
==== Front
Appl Microbiol Biotechnol
Appl Microbiol Biotechnol
Applied Microbiology and Biotechnology
0175-7598
1432-0614
Springer Berlin Heidelberg Berlin/Heidelberg

39264460
13282
10.1007/s00253-024-13282-4
Biotechnological Products and Process Engineering
Exploration of diverse secondary metabolites from Penicillium brasilianum by co-culturing with Armillaria mellea
Rong Xiaoting 1
Zhang Lihua 2
He Wenni 1
Guo Zhe 1
Lv Hui 1
Bai Jinglin 1
Yu Liyan yly@cpcc.ac.cn

1
Zhang Lixin lxzhang@ecust.edu.cn

3
http://orcid.org/0000-0002-0715-1739
Zhang Tao tzhang_0218@163.com

1
1 https://ror.org/02drdmm93 grid.506261.6 0000 0001 0706 7839 Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100050 China
2 grid.410648.f 0000 0001 1816 6218 National Key Laboratory of Chinese Medicine Modernization, State Key Laboratory of Component-Based Chinese Medicine, Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617 China
3 grid.28056.39 0000 0001 2163 4895 State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai, 200237 China
12 9 2024
12 9 2024
2024
108 1 46221 4 2024
7 8 2024
14 8 2024
© The Author(s) 2024
2024
https://creativecommons.org/licenses/by-nc-nd/4.0/ Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
Abstract

Bioinformatic analysis revealed that the genomes of ubiquitous Penicillium spp. might carry dozens of biosynthetic gene clusters (BGCs), yet many clusters have remained uncharacterized. In this study, a detailed investigation of co-culture fermentation including the basidiomycete Armillaria mellea CPCC 400891 and the P. brasilianum CGMCC 3.4402 enabled the isolation of five new compounds including two bisabolene-type sesquiterpenes (arpenibisabolanes A and B), two carotane-type sesquiterpenes (arpenicarotanes A and B), and one polyketide (arpenichorismite A) along with seven known compounds. The assignments of their structures were deduced by the extensive analyses of detailed spectroscopic data, electronic circular dichroism spectra, together with delimitation of the biogenesis. Most new compounds were not detected in monocultures under the same fermentation conditions. Arpenibisabolane A represents the first example of a 6/5-fused bicyclic bisabolene. The bioassay of these five new compounds exhibited no cytotoxic activities in vitro against three human cancer cell lines (A549, MCF-7, and HepG2). Moreover, sequence alignments and bioinformatic analysis to other metabolic pathways, two BGCs including Pb-bis and Pb-car, responsible for generating sesquiterpenoids from co-culture were identified, respectively. Furthermore, based on the chemical structures and deduced gene functions of the two clusters, a hypothetic metabolic pathway for biosynthesizing induced sesquiterpenoids was proposed. These results demonstrated that the co-culture approach would facilitate bioprospecting for new metabolites even from the well-studied microbes. Our findings would provide opportunities for further understanding of the biosynthesis of intriguing sesquiterpenoids via metabolic engineering strategies.

Key points

• Penicillium and Armillaria co-culture facilitates the production of diverse secondary metabolites

• Arpenibisabolane A represents the first example of 6/5-fused bicyclic bisabolenes

• A hypothetic metabolic pathway for biosynthesizing induced sesquiterpenoids was proposed

Graphical Abstract

Supplementary Information

The online version contains supplementary material available at 10.1007/s00253-024-13282-4.

Keywords

Co-culture
Penicillium brasilianum
Armillaria mellea
Secondary metabolites
Sesquiterpenes
http://dx.doi.org/10.13039/501100001809 National Natural Science Foundation of China 31872617 Zhang Tao http://dx.doi.org/10.13039/501100019018 Chinese Academy of Medical Sciences Initiative for Innovative Medicine 2021-I2M-1-055 2019-I2M-1-005 Zhang Tao issue-copyright-statement© Springer-Verlag GmbH Germany, part of Springer Nature 2024
==== Body
pmcIntroduction

Filamentous fungi have been demonstrated to be an important source of agrochemicals, pharmaceutical drugs, and other chemical agents (Bills and Gloer 2016; Keller 2019; Zhang et al. 2023b, 2013). However, it is becoming increasingly difficult to screen novel lead compounds by conventional chemical-only strategies for high frequency of repeated discovery of known metabolites (Malit et al. 2022). The fact that the metabolic potential has not yet been exploited is due to the observation that most gene clusters are transcriptionally silent under normal laboratory culture conditions. Furthermore, genome sequencing has revolutionized secondary metabolites (SMs) mining efforts, manifesting that the capacity of filamentous fungi to produce compounds was far more than we anticipated previously (Keller 2019; Medema et al. 2021). To circumvent these limitations, several strategies have been conducted to trigger cryptic biosynthetic pathways, exemplified as “one strain many compounds (OSMAC)” approach (Bode et al. 2002; Pan et al. 2019; Zhang et al. 2022a), metabolic engineering of targeted pathways (Zhang et al. 2022b, 2018), epigenetic modification (Cichewicz 2010; Mao et al. 2015; Zheng et al. 2022), heterologous expression of gene clusters using different host (Lyu et al. 2021; Oikawa 2020; Zhang et al. 2023a), and co-culture (Bertrand et al. 2014; Wang et al. 2022; Zhuang and Zhang 2021). Previous studies have demonstrated that co-culture methodology is effective in facilitating the discovery of new SMs with fascinating structures (Knowles et al. 2022; Park et al. 2017). Thus, the genome-guided co-culturing approach renders the isolation of bioactive molecules efficient and promising.

Cell–cell interactions of different microorganisms could simulate the competition of natural ecosystems (Ghoul and Mitri 2016; Weiland-Brauer 2021), and the cryptic metabolic pathways are transcriptionally triggered by interspecies crosstalk or environmental stimuli (Zhang et al. 2022a). The presence of another strain population might stimulate strain behavior or culturing success (Goers et al. 2014). Also, the chemical inducer released from other microbes might cause defense responses, which include yielding diverse arrays of SMs, secretion of extracellular enzymes, the synergistic act of biotransformation, or signaling pathways activation (Bouws et al. 2008; Elhamouly et al. 2022). Co-culture systems are highly relevant and could find myriad applications in mining metabolites based on the natural interactions between cell populations. Recently, the microbial co-culture-based SMs mining approach has attracted increasing interest, as exemplified by bacterial-bacterial (Sugiyama et al. 2019), fungal-bacterial (Park et al. 2017), and fungal-fungal co-cultivation systems (Wang et al. 2018). These studies highlighted the capability of co-culture to effectively trigger the generation of metabolites (Kim et al. 2021). Thus, the different co-culture systems have increased productivity over monocultures and hold greater potential for mining novel compounds.

Penicillium is a diverse genus occurring worldwide composed of 354 accepted species (Visagie et al. 2014), and a large number of SMs exhibiting potent bioactivities and intriguing structural features are reported from Penicillium genus, such as polyketides, alkaloids, peptides, terpenoids, and hybrids (Brase et al. 2009; Kozlovsky et al. 2020). P. brasilianum could produce compounds including griseofulvin, verruculogen, brasiliamides, and meroterpenoids, and has been an interesting fungus to many researchers in the continuous screening for new compounds (Bazioli et al. 2017). Metabolomic profiling of Armillaria on rice medium was previously performed indicating that only a small subset of metabolites could be produced (Zhang et al. 2022a). Additionally, genomic analysis further revealed a rich repertoire of genes in Armillaria encoding cytochrome P450s and plant cell wall–degrading enzymes (PCWDEs) genome (Zhang et al. 2022a, 2023a). Therefore, a scaled-up fermentation of the co-culture system composed of P. brasilianum and the A. mellea was performed. In the present study, systematic chemical investigation enabled the isolation of five new compounds, including two bisabolene-type sesquiterpenes, named arpenibisabolanes A and B (1–2), two carotane-type sesquiterpenes, named arpenicarotanes A and B (3–4), and one polyketide, arpenichorismite (5), together with seven known compounds. Notably, some of the new compounds (1–5) were not produced by either of the two fungi when cultivated alone under the same fermentations, and arpenibisabolanes A (1) represents the first example of a 6/5-fused bicyclic bisabolene. Herein, we described the isolation and structural elucidation of these metabolites. The BGCs and hypothetical metabolic pathways for biosynthesizing induced sesquiterpenoids were also investigated.

Materials and methods

Fungal strains and culture conditions

The fungi P. brasilianum CGMCC 3.4402 (NBRC 6234) and A. mellea CPCC 400891 (DSM 3731) used in this study were deposited in the China Pharmaceutical Culture Collection Center (CPCC). The fungi were routinely cultivated on potato dextrose medium (PDA) and preserved in 15% glycerol at − 80 °C. The growth temperature of the strains is 28 °C. OSMAC-based evaluation of strain NBRC 6234 producing secondary metabolites was conducted using eight different culture media as performed previously: PDB broth, YMEG broth, MEP broth, F1 broth, F2 broth, CY broth, and rice medium (20 g rice/20 mL of deionized water in 250-mL Erlenmeyer flask) (Zhang et al. 2022a). The fermentation cultures include a two-stage process; the fungus P. brasilianum was cultivated on PDA media at 28 °C for 7 days and served as seed culture. The spores were harvested and inoculated equally into the fermentation medium (100 mL respective broth in 500-mL Erlenmeyer flasks) described above. The routine fermentation was cultivated for 10 days on a shaker (150 r/min) or incubated statically in rice medium for 25 days (Rong et al. 2023a, 2023b).

Sequencing and bioinformatic tools

Genomic DNA (gDNA) from Penicillium was extracted from mycelia cultivated in PDB broth using E.Z.N.A.® Fungal DNA Mini Kit (Omega, Norcross, GA, USA) according to the manufacturer’s instruction. Genome sequencing was conducted at Shanghai Majorbio Bio-pharm Technology Co. Ltd. (Shanghai, China) using the Illumina Hiseq 2000 platform. The contigs that were assembled and annotated by SOAPdenovo 1.05 were formatted to the UniProt database for BLAST alignment (Bateman et al. 2023; Luo et al. 2012). The AntiSMASH software was used for analysis of secondary metabolites biosynthetic gene clusters (BGCs) (Blin et al. 2019). Gene annotations of the coding sequences were deduced and verified manually based on the homologues in the NCBI database.

Co-culturing approach and large-scale fermentation

For the fungus A. mellea, the cultivation was performed in MEP broth based on the details as previously reported (200 mL each in 500-mL Erlenmeyer flasks × 5) for 10 days (Zhang et al. 2022a, 2023a). For the Penicillium strain CGMCC 3.4402, it was cultivated in PDB broth (200 mL each in 500-mL Erlenmeyer flasks × 3) for 7 s days. The large-scale co-culture fermentation (6 kg rice) was carried out in Erlenmeyer flasks (100 g rice/100 mL of deionized water in 500-mL Erlenmeyer flask). The fungal strains were transferred into rice media simultaneously, in which the inoculants of Penicillium and Armillaria were 5 mL, respectively. The fermentations were cultured at 28 °C under static conditions for 30 days (Zhang et al. 2022b).

Chemicals and chemical analysis

Circular dichroism spectra were recorded using a Jasco J-815 spectropolarimeter (Jasco, Tokyo, Japan). High-performance liquid chromatography (HPLC) analyses were carried out on an Agilent 1290 instrument with a YMC-Pack ODS-AQ column (4.6 × 250 mm, 5 µm). TLC was conducted on silica gel GF254 plates from Qingdao Marine Chemical, China. The medium-pressure liquid chromatography was carried out on Combi Flash Rf 200 (Teledyne Isco, Lincoln NE, USA) using a SEPAF FLASH® Flash silica gel column (40–63 µm, 60 Å, 330 g, Santai Technologies, China). The semi-preparative HPLC separations were performed on SSI series 1500 (Cometro Technology Ltd, NJ, USA) equipped with a DAD detector using a YMC-Pack ODS-AQ column (10.0 mm × 250 mm, 5 µm). High-resolution mass spectrometric data was obtained using a Thermo LTQ Orbitrap XL Mass Spectrometer installed with an electrospray ionization source (Thermo Fisher Scientific, CA, USA). Nuclear magnetic resonance (NMR) data were acquired on a Bruker AVIII-600 spectrometer using TMS as internal standard (150 MHz for 13C NMR and 600 MHz for 1H NMR, Bruker Corporation, Germany), which were measured in CDCl3 or CD3OD.

Extraction and isolation of secondary metabolites

After the fermentation period, the cultures of different fermentation media were centrifuged to harvest the mycelia from broth, respectively (5500 r/min, 15 min). The supernatant and respective fungal material were extracted twice with ethyl acetate (EtOAc) separately and successively combined to afford one sample (Zhang et al. 2022a). The culture of rice medium was directly extracted using EtOAc twice. All seven solutions were collected and concentrated in vacuo to dryness. Each crude extract was redissolved in MeOH and analyzed following the same methodology described previously (Rong et al. 2023a, 2023b).

The co-culture materials were extracted repeatedly with MeOH (3 × 25 L), and the successive MeOH extracts were combined and evaporated under vacuum yielding 300 mL of crude extracts. The materials were fractionated with n-hexane (3 × 200 mL) and EtOAc (4 × 500 mL), and the EtOAc solvent was evaporated to dryness under rotatory evaporation to yield the crude extract (ca. 22 g). The crude extract was subsequently separated on silica gel (200–300 mesh) column chromatography (12 cm × 40 cm). The column was eluted with a gradient consisting of CHCl2/MeOH (v/v, 100:0 → 95:5 → 90:10 → 85:15 → 80:20 → 75:25 → 70:30 → 65:35 → 60:40 → 55:45 → 50:50 → 45:55 → 40:60 → 0:100), to obtain 10 fractions (Fr.1–Fr.10). Fr.6 was further separated by ODS column, using a stepped gradient elution of CH3CN-H2O (20:80 to 100:0) to yield 14 subfractions (Fr.6–1 to Fr.6–14). Fraction Fr.6–2 was purified by pHPLC eluted with CH3CN-H2O (0.01% TFA) (20:80) to afford compound 9 (6.8 mg, tR = 8.0 min). Purification of fraction Fr.6–9 was conducted by semi-preparative HPLC, eluting with CH3CN-H2O (0.01% TFA) (25:75) to afford compounds 3 (4.6mg, tR = 25.8 min) and 7 (3.5 mg, tR = 26.8 min). Fraction Fr.6–3 was separated by semi-preparative HPLC with CH3CN-H2O (0.01% TFA) (10:90) to yield compounds 12 (3.5 mg, tR = 11.0 min), 10 (3.5 mg, tR = 18.0 min), and 11 (4.8 mg, tR = 20.0 min). Fraction Fr.6–5 was purified by pHPLC eluted with CH3CN-H2O (0.01% TFA) (40:60) to obtain compound 1 (4.5 mg, tR = 13.0 min). Purification of fraction Fr.6–5 was conducted by semi-preparative HPLC, eluting with CH3CN-H2O (0.01% TFA) (40:60) to afford compound 2 (4.5 mg, tR = 13.0 min). Purification of fraction Fr.6–6 was performed by pHPLC, using CH3CN-H2O (0.01% TFA) (40:60) to obtain compound 4 (7.2 mg, tR = 7.0 min) and compound 6 (5.7 mg, tR = 5.4 min). Fraction Fr.5 was eluted with a gradient using a silica gel column CH3CN-H2O (20:80 to 100:0) to yield 13 subfractions (Fr.5–1 to Fr.5–13). Fraction Fr.5–2 was purified by semi-preparative HPLC with CH3CN-H2O (0.01% TFA) (15:85) to obtain compound 8 (14.4 mg, tR = 25.0 min). Fraction Fr.5–4 was purified by pHPLC eluted with CH3CN-H2O (0.01% TFA) (25:75) to acquire compound 5 (10.9 mg, tR = 15.0 min).

Electronic circular dichroism (ECD) calculation of 1 − 4

Stochastic conformational searches were firstly conducted under MMFF94 force field for 1 − 4 and gave their conformers. Their conformers were optimized at the B3LYP/6-31G(d) basis set level in methanol, and the frequencies were calculated by further time-dependent density functional theory (TDDFT) method, indicating that these conformers were stable. Using the conformers at the B3LYP/6-31G(d) basis set level in methanol, their excitation states at the B3LYP/6-31G(d) basis set level were calculated, peak stretcher was 0.4 eV, and finally the calculation results were Boltzmann averaged to yield the depicted electronic circular dichroism (ECD) spectra of 1 − 4. All calculations were performed by Gaussian 09 program package (Version C.01).

Cytotoxic activity assays

Five new metabolites were tested for their cytotoxic activities as previously reported methods (Rong et al. 2023a, 2023b; Zhang et al. 2022a). Briefly, cells (3 × 104 cells/mL) were inoculated in the wells of a 96-well plate at 100 µL/well. After 24 h of incubation at 37 °C and 5% CO2, cells were cultivated to 90% confluence. Carcinogenic cell lines including A549, MCF-7, and HepG2 were treated with gradient concentrations of isolated compounds for 48 h (DMSO and cisplatin were used as negative and positive controls, respectively). The concentration gradient of cisplatin is 20, 10, 5, 2.5, 1.25, and 0.625 µM, respectively. The cytotoxicity was measured by the CCK-8 method (Zhang et al. 2022a). Measure the absorbance at 450 nm using a microplate reader (Bio-Rad, Fitchburg, WI, USA). The dilutions of the tested compounds were independently performed thrice. The half-maximal inhibitory concentration (IC50) of compounds in three cell lines was calculated using GraphPad Prism 7. Purities of tested metabolites were > 95% detected by HPLC-ELSD.

Gene Pb_bisA synthesis, yeast transformation, and GC–MS analysis

The candidate gene Pb_bisA was synthesized by GenScript and codon-optimized for expression in yeast. The oligonucleotide primers used for are listed in Table S1 (Supporting information). PCR reaction is performed using 2 × Phanta Max Master Mix (Vazyme) following the manufacturer’s instructions. Yeast expression plasmid YET was used for construction of YET-Pb_bisA by in vivo homologous recombination strategy. Yeast transformation was conducted using a Frozen-EZ Yeast Transformation II Kit™ (Zymo Research). Yeast plasmid was prepared by a E.Z.N.A® Yeast Plasmid Miniprep Kit (Omega) and transformed into E. coli Trans-T1 (TransGen Biotech) for sequencing.

The fermentation and samples preparation for gas chromatography–mass spectrometry (GC–MS) analysis were performed as reported previously with minor modifications (Li et al. 2023b; Zhang et al. 2023a). The 5.0 mL cultures of yeast transformants were collected in the headspace (20 mL), followed by incubation at 55 °C for 20 min. The volatile components were harvested at 55 °C for 30 min by solid-phase micro-extraction (SPME) with a DVB/CAR/PDMS fiber. The samples were analyzed on an Agilent 7890B/5977A gas chromatography incorporated with an Agilent 7200 accurate-mass quadrupole time-of-flight (GC–MS-TOF) and a DB-WAX column (Agilent, 60 m × 0.25 mm ID, 0.25 µm film thickness). The system and the program were performed under the standard condition described previously (Li et al. 2023b; Zhang et al. 2023a). Full-scan mass spectra were recorded in the range of 100–300 m/z, and ionization was conducted by electron impact at 70 eV with an electrospray ionization source temperature adjusted at 230 °C.

Results

Constructing co-culture system to induce SMs production

To investigate the biosynthetic potential of P. brasilianum, we initially performed a whole genome sequencing of the fungus. The Illumina HiSeq 2500 sequencing of the strain yielded a total of ~ 5148 million bases. Assembling of the unpaired reads generated 44 scaffolds, which consist of 34.12 million nonredundant bases. During the antiSMASH-guided bioinformatic analysis, forty BGCs were obtained and the potential biosynthetic compounds include polyketides (no. 12), nonribosomal peptides (no. 17), hybrids (no. 4), terpenes (no. 5), and beta lactones (no. 2). Subsequently, OSMAC approach was applied to evaluate the chemical diversity of the strain. However, the fungus did not biosynthesize many secondary metabolites in the rice medium, PDB broth, MEP broth, F1 broth, F2 broth, YMEG broth, or CY broth (Fig. S1, supporting information), which are commonly used for promoting chemodiveristy in filamentous ascomycetes (Ariantari et al. 2019; Zang et al. 2020). These results indicated that the OSMAC strategy could not broaden the structurally diverse pattern of SMs from the strain P. brasilianum.

The A. mellea exhibits a parasitic lifestyle and is widely distributed as devastating pathogens of herbaceous plants or broadleaf trees (Baumgartner et al. 2011; Fradj et al. 2020). Significantly, the Armillaria encode a large number of PCWDEs (Zhang et al. 2023a). Thus, this strain was believed to be capable of generating interaction with the Penicillium and served as an inducing strain of co-culture system. As a control, the culture of Armillaria strain alone resulted in a similar condition (Fig. 1, trace I; Fig. S2, supporting information). Markedly, chemical analysis of the strain P. brasilianum cultivated in rice medium resulted in a sparse metabolite profile, only a few dominating peaks were visible in the HPLC-ELSD chromatogram (Fig. 1, trace ii; Fig. S2, supporting information). However, the co-culture significantly stimulated the expression of BGCs and/or uncharacterized metabolic pathways (Fig. 1, trace iii; Figs. S2 and S3, supporting information). This demonstrates that the co-culture system is feasible, which could stimulate the production of new or uncharacterized SMs.Fig. 1 HPLC profiles of organic extracts obtained from the strain P. brasilianum by co-culture strategy. (i) A. mella; (ii) P. brasilianum; (iii) co-culture. The extracts were analyzed by measuring UV absorbance spectra at 230 nm on an Agilent 1290 Infinity system equipped with an Alltech ELSD 2000 detector

Identification of induced metabolites from co-culture system

Compound 1 was obtained as colorless oil. The molecular formula C15H18O6, with seven degrees of unsaturation, was determined by HR-ESI–MS at m/z 293.1020 [M-H]− (calcd for C15H17O6− 293.1025). The 1D NMR (Table S2, supporting information) and HSQC data of 1 clearly displayed the presence of two carbonyls [δC 189.5 and 171.4], three double bonds [δC/δH 148.3/7.69 (1H, s), 141.9/6.78 (1H, tq, J = 7.44, 1.44 Hz); δC 141.7, 138.1, 130.2, 126.9], two methyls [δC/δH 12.5/1.76 (3H, s), 18.2/1.30 (3H, s)], three methylenes [δC/δH 29.5/2.48 (2H, m); 28.4/3.06 (1H, dd, J = 5.4, 16.8 Hz), 2.58(1H, m); 22.8/2.62(2H, m)], and two oxygenated quaternary carbons [δC 79.3, 75.2] (Fig. 2). The key HMBC correlations (Fig. 3) of H3-15/C-2, C-3, C-4; of H-2/C-3, C-6; of H-1/C-2, C-5, C-6, C-7; and H-14/C-5, C-6, C-7 led to the assignment of a bicyclic system consisted of a cyclohexanone ring fused with a furan ring through C-5 and C-6. Then, a C5 α, β-unsaturated carboxylic acid side chain at C-7 could be inferred, supported by the molecular formula of 1 and the key HMBC correlations (Fig. 3) of H-13/C-10, C-11, C-12; of H-10/C-8, C-9, C-11, C-12; of H-9/C-7, C-8, C-11; and of H-8/C-1. Thus, the gross structure of 1 was established. Due to no available NOESY spectrogram, the relative configuration of C-2/C-3 could not be established. Further, the absolute configuration of 1 was assigned as 2S,3R by comparison of the experimental and calculated ECD spectra of four configurations (2R,3R-1a, 2S,3S-1b, 2S,3R-1c, 2R,3S-1d) (Fig. 4A and B). Accordingly, the absolute configuration of 1 was determined and characterized as arpenibisabolane A (Figs. S4–S11, supporting information).Fig. 2 Structures of compounds 1–12 isolated in the study

Fig. 3 Key HMBC, COSY, and NOESY correlations of 1–5. A Key HMBC and COSY correlations of 1–5. B NOESY correlations of 2–4

Fig. 4 Calculated and experimental ECD spectra for compounds 1–4. A, B Arpenibisabolane A (1). C Arpenibisabolane B (2). D Arpenicarotane A (3). E, F Arpenicarotane B (4) in MeOH

The molecular formula of compound 2 was assigned as C15H20O4 based on the HR-ESI–MS at m/z 263.1278 [M-H]− (calcd for C15H19O4− 263.1283), corresponding to six degrees of unsaturation. Comparison of the 1D NMR data (Table S2, supporting information) of 2 with those of 1 indicated that 2 shared the identical α, β-unsaturated carboxylic acid moiety. The remaining NMR data (Table S2, supporting information) corresponded to one carbonyl (δC 192.7), two double bonds [δC/δH 129.5/5.84 (1H, m); δC 164.3, 129.3, 149.2], two methyls [δC/δH 21.0/1.94 (3H, s), 20.2/2.02 (3H, s)], one methylene [δC/δH 38. 0/3.0 (1H, m), 2.61 (1H, m)], and one oxygenated methine [δC/δH 70.0/4.25 (1H, m)]. The key HMBC correlations (Fig. 2) of H-13/C-10, C-11, C-12; of H-10/C-8, C-12; of H-9/C-11; and of H-14/C-7, C-8 confirmed the presence of a C6 α, β-unsaturated carboxylic acid side chain (C-7–C-12). Then, one α, β-unsaturated cyclohexanone moiety was deduced by the key HMBC correlations of H3-15/C-2, C-3, C-4; of H-2/C-4, C-6; and H2-5/C-1, C-3, C-4. Further, HMBC correlations of H3-14/C-6 and H2-5/C-7 led to the linkage of C-6/C-7 (Fig. 3). Thus, the gross structure of 2 was established (Fig. 2). The E-geometry of Δ10(11) and Z-geometry of Δ6(7) double bonds were assigned by the NOESY correlations (Fig. 3) of H3-14/H2-5 and H3-13/H2-9. The absolute configuration of 2 was determined by comparison of its experimental and calculated ECD data. The experimental ECD curve (Fig. 4C) of 2 agreed well with the calculated curve of 4S-2a, which assigned its absolute configuration as 4S. Thus, the structure of 2 was established and named arpenibisabolane B (Figs. S12–S20, supporting information).

Compound 3 has a molecular formula of C15H24O4, which was deduced from its HR-ESI–MS data at m/z 291.1550 [M + Na]+ (calcd for C15H24O4Na+ 291.1550), corresponding to four degrees of unsaturation. The 1H NMR data (Table S3, supporting information) exhibited one olefinic proton at δH 7.07 (1H, m), two oxygenated methylenes at δH 3.40 (1H, d, J = 11.1 Hz) and 3.36 (1H, d, J = 11.4 Hz), and two methyls at δH 1.12 (3H, s) and 0.78 (3H, s). The 13C NMR (Table S3, supporting information) and HSQC data revealed the existence of fifteen carbon signals, including one carbonyl, two sp2 carbons, two methyls, five methylenes, three methines, and two quaternary carbons (Fig. 2). The key HMBC correlations (Fig. 3) of H3-15/C-2, C-7; of H2-2/C-1, C-3, C-4; of H-3/C-5, C-14; of H2-5/C-6, C-14; of H2-6/C-1, C-4; of H-7/C-1, C-2, C-5, C-6, C-8; of H-10/C-1, C-7, C-8, C-9, C-15; and of H-9/C-8 suggested the presence of a bicyclic system consisted of a cyclopentane fused with a cycloheptane with one methyl and one carboxyl respectively assigned at C-15 and C-4. Then, the key HMBC correlations (Fig. 3) of H3-13/C-8, C-11; of H2-12/C-8, C-11, C-13; of H2-9/C-11; of H-7/C-11; of H3-13/C-11, C-8; of H2-12/C-8 and C-11, C-13; and of H2-9/C-11, and H-7/C-11, together with the downfield chemical shifts of C-11 (δC 76.8) and C-12 (δC 70.6), assigned a dihydroxy-isopropyl group at C-8. Consequently, the gross structure of 3 was established. The NOESY correlations (Fig. 3) of CH3-15/H-8/H2-12 and CH3-13/H-7 indicated that H-8, H2-12, and H3-15 were β-oriented, and H3-13 and H-7 were α-oriented. By comparison of the experimental and calculated ECD spectra (Fig. 4D), the absolute configuration of 3 was assigned as 1R,7S,8R,11R. Thus, the structure of 3 was established and named arpenicarotane A (Fig. S21–S28, supporting information).

Compound 4 was deduced to possess the molecular formula of C15H22O4 by analysis of its HR-ESI–MS data at m/z 265.1439 [M-H]− (calcd for C15H21O4− 265.1440), corresponding to five degrees of unsaturation. The HSQC correlations and 1D NMR data (Table S3, supporting information) of 3 showed the presence of one carbonyl [δC 176.4], two double bonds [δH/δC 4.83 (1H, s), 4.73 (1H, s)/111.9, 5.57 (1H, q, J = 2.4 Hz)/126.1; δC 149.2, 149.1], two quaternary carbons [δC 79.3, 44.4], two methyls [δH/δC 1.83 (3H, s)/22.3, 1.06 (3H, s)/20.9], two methines [δH/δC 2.79 (1H, d, J = 12.7 Hz)/51.2, 3.72 (1H, dd, J = 11.6, 5.2 Hz)], and four methylenes [δH/δC 2.31 (1H, m), 1.29 (1H, m)/22.5; 2.23(1H, m), 1.76 (1H, m)/33.8; 2.08 (1H, dd, J = 13.0, 5.2 Hz), 1.98 (t, J = 13.0 Hz)/47.4; 2.15 (1H, d, J = 16.0 Hz), 2.04 (1H, dd, J = 16.0, 2.4 Hz)/49.4] (Fig. 2). In the key HMBC correlations (Fig. 3) of H3-15/C-1, C-6, C-7, C-8; of H2-6/C-1, C-4, C-5; of H-5/C-4, C-14; of H2-3/C-1, C-4, C-5, C-14; of H2-2/C-1, C-4, C-7; of H-9/ C-1, C-7, C-8, C-10; and of H2-8/C-15, C-9, combined with the 1H-1H COSY correlations of H-5/H2-6, H-10/H-1, H-1/H2-2 and H2-2/H2-3, a bicyclic system consisted of a cyclopentane fused with a cycloheptane with a methyl located at C-15 and a carboxyl assigned at C-4 could be deduced. Considering the downfield chemical shifts of C-4 (δC 79.3) and C-5 (δC 77.5), two hydroxy groups at C-4 and C-5 could be inferred. Further, HMBC correlations of H2-13/C-12, C-11 and of H3-13/C-10, C-11 established an isopropenyl positioned at C-10. Thus, the planar structure of 4 was established. The NOESY (Fig. 3) correlations of H-13/H-2 and CH3-12/H-9 indicated the trans-form for the double bonds Δ9(10) and Δ11(13). The NOESY correlations of CH3-15/H-5/H-6 (δH 1.98) and H-1/H-6 (δH 2.08) indicated that CH3-15 and H-5 were β-oriented, while H-1 and HO-5 were α-oriented. The relative configuration of C-4 could not be determined due to a lack of available NOESY correlations. Finally, comparison of the experimental and calculated ECD spectra (Fig. 4E and F) assigned the absolute configuration of 4 as 1R,4R,5S,7R. Thus, the structure of 4 was determined and named arpenicarotane B (Fig. S29–S37, supporting information).

The molecular formula of compound 5 was deduced to be C11H10O6 with nine degrees of unsaturation, based on the analysis of its HR-ESI–MS data at m/z 237.0400 [M-H]− (calcd for C11H9O6237.0405). The 1H NMR data (Table S4, supporting information) showed a 3H 1,3,5-trisubstituted benzene system at δH 7.19 (1H, t, J = 1.8 Hz), 7.08 (1H, t, J = 1.8 Hz), and 6.28 (1H, t, J = 2.3 Hz), two geminal olefinic protons at δH 5.81 (1H, d, J = 1.68 Hz) and 5.10 (1H, d, J = 1.68 Hz), and a methoxy group at δH 3.87 (3H, s). The 13C NMR data (Table S4, supporting information) showed the existence of eleven carbon signals, including two carbonyls, eight sp2 carbons, and one methoxy carbon, which were unambiguously designated by the HSQC experiment (Fig. 2). HMBC correlations (Fig. 3) of H-6/C-2, C-4, C-5, C-7; of H-4/C-3, C-5, C-6; of H-2/C-3, C-4, C-7; and of H3-8/C-7 confirmed the presence of 1,3,5-trisubstituted aromatic ring with one hydroxyl and one methoxycarbonyl group positioned at C-5 and C-1, respectively. Further, the key HMBC correlations of H2-11/C-9 and C-10, as well as the downfield chemical shifts of C-3 (δC 158.8), were indicative of a monosubstituted acrylic acid moiety connected to C-3 of the benzene ring through an ether bond. Thus, compound 5 was identified as a new chorismite derivate and named arpenichorismite A (Fig. S38–S44, supporting information).

The remaining known compounds were identified as aspterric acid (6) (Xing et al. 2019), aspergerthinacid A (7) (Pan et al. 2021), 8-O-4-coupled dehydrodiferulic acid (8) (Ralph et al. 1994), methy 4-hydroxyphenylacetate (9) (Shen et al. 2013), 4-hydroxyphenylacetic acid (10) (Ohtani et al. 2011), p-tolyl-3-aminopropanoate (11) (Xie et al. 2008), and p-(acetylamino) benzoic acid (12) (Lewis et al. 2003) by comparison of their NMR spectral data and optical rotation values with those reported in the literature (Fig. S45–S64, supporting information).

Bioinformatic analysis of the BGCs Pb_bis and Pb_car

Further detailed investigation of the structures of the metabolites (1–4) isolated from the co-culture, two different classes of sesquiterpenoids including bisabolene- and carotane-type were characterized. To obtain the encoding gene clusters that potentially might be involved in the biosynthesis of arpenibisabolanes and arpenicarotanes, we examine the genome sequences of P. brasilianum and the basidiomycete A. mellea (https://mycocosm.jgi.doe.gov/Armme1_1/Armme1_1.home.html) (Li et al. 2023b). Interestingly, three sesquiterpenoids including compounds 2, 4, and 6 were also isolated from the genetically modified mutant derived from the strain P. brasilianum, in which a pathway-specific activator BerA involving berkeleyacetals biosynthesis from Neosartorya glabra was overexpressed (personal communication). This implied that the P. brasilianum possessed the capability to biosynthesize the bisabolene- and carotane-type sesquiterpenoids. The biosynthetic locus of aspterric acid has been previously characterized in Aspergillus terreus NIH2624; thus, we searched for genomic locus coding enzymes resembling to those encoded by ast cluster. The cluster Pb_car was ascertained and it is highly homologous to ast cluster in the strain A. terreus and both clusters exhibited syntenic and highly conserved (Fig. 5A, Table S5, supporting information). Bioinformatic analysis of the cluster Pb_car allowed the discovery of genes coding typical enzymes (Fig. 5B), which include the scaffold-forming terpene cyclase (CarA), two post-modification cytochrome P450s (CarB and CarC), and one dihydroxyacid dehydratase (CarD) responsible for self-tolerance.Fig. 5 Bioinformatic analysis of biosynthetic clusters involved in formation of arpenibisabolanes and arpenicarotanes. A Phylogenetic tree of STSs from P. brasilianum, A. mella, and closely characterized α-bisabolol synthases using the Neighbor-Joining method. The scale shows changes per site; numbers at branches are bootstrap values. Pb-CarA and Pb-BisA are STSs from P. brasilianum, Arm_STS1-Arm_STS14b are STSs from A. mellea. B Putative biosynthetic clusters of isolated metabolites. Pb_bis is the encoded cluster of arpenibisabolanes, Pb_car is the encoded cluster of arpenicarotanes

Arpenibisabolanes (1–2) are sesquiterpene-derived and the scaffold should be biosynthesized by the sesquiterpene synthase (STS). Therefore, all STSs encoded from two fungal strains were recovered, in which eighteen STSs from A. mellea and two hypothetical sesquiterpene synthases from the strain P. brasilianum were included. A phylogenetic dendrogram of STSs obtained according to bioinformatic mining from co-culture and previously characterized α-bisabolol synthases was constructed, exhibiting a clear separation of terpene synthases (Fig. 5A). In this phylogenetic tree, one synthase designated Pb_BisA formed a distinct branch with UbiA-type α-bisabolol synthases (BibS) from Fusarium sp. JNU-XJ070152 and Stachybotrys sp. PYH05-7 (Luo et al. 2022), β-trans bergamotene synthase from Aspergillus fumigatus Af293 (Lin et al. 2013) and plant-derived alpha-bisabolol synthases exemplified as SspiBS from Santalum spicatum (Jones et al. 2011), or homologues from Arabidopsis thaliana and Artemisia annua (Muangphrom et al. 2016; Ro et al. 2006). This indicates it might support the proposal of Pb_BisA as a crucial synthase in arpenibisabolanes biosynthesis. DNA sequence analysis of the clustered ~ 7.5 kb Pb_bis locus (Fig. 5B), and further amino acid alignment revealed that Pb_BisA partakes of 36% identity to alpha-bisabolol synthases from Stachybotrys sp. PYH05-7, followed by BibS from Fusarium sp. JNU-XJ070152-01 (31% identity) and bergamotene synthase from A. fumigatus Af293 (24% identity). In addition to Pb-BisA, neighboring genes encoded the cytochrome P450 (BisB), G-protein coupled receptors (GPCR)–like protein (BisC), and decarboxylase (BisD) were also identified. To characterize that the target Pb_BisA is responsible for the production of arpenibisabolanes, the Pb_bisA was heterologously expressed in yeast Saccharomyces cerevisiae BJ5464. Then, a combination of GC–MS analysis with National Institute of Standards and Technology (NIST) Standard Reference Database (v20, 2023–01, https://webbook.nist.gov/) searching revealed that the compound β-bisabolene (m/z = 204, C15H24) could be detected and identified (Fig. S65, supporting information). This reinforced that the target gene cluster Pb_bis involved in arpenibisabolanes biosynthesis.

Hypothetic biogenesis pathway of induced sesquiterpenes

For compounds 1 and 2, nevertheless, only a few studies have been carried out for scrutinizing the metabolic process of such skeletal unique molecules. Luo et al. characterized the α-bisabolol synthases FmaTC and StaTC1 from Fusarium sp. and Stachybotrys sp., respectively (Luo et al. 2022). These UbiA-type terpene cyclases could produce diverse bisabolene-derived molecules and the pioneering biogenesis insights of bisabolol derivatives were only preliminary. Although compounds 1 and 2 displayed different structures, their primary skeletons are bisabolene-derived. To explain the possible biogenetic origin, their plausible metabolic pathways were proposed in Scheme 1A. Arpenibisabolanes (1–2) initiate from farnesyl pyrophosphate (FPP) and the enzymatic cyclization generates the intermediate β-bisabolene. Nevertheless, considering the isolated 1 and 2 that exhibit a wealth of oxygenated modifications, we preferred that a multi-step oxidative cascade occurs after the bisabolene formation step. These conversions might involve one cytochrome P450 monooxygenase (BisB)–catalyzed oxidations. The BisB exhibits a relatively higher identity to the cytochrome P450 (Af510), which catalyzed a cascade of oxidations during fumagillin generation from Aspergillus fumigatus (Lin et al. 2013). This implies that BisB might be multifunctional and it was believed to play a key role in generating highly oxygenated structure during arpenibisabolanes biosynthesis. In this scheme, the generation of compound 1 might be modified by auxiliary oxygenases encoded by Armillaria species.Scheme 1 Proposed metabolic pathway of arpenibisabolanes and arpenicarotanes biosynthesis

Biogenetically, compounds 3, 4, and 6 possess a unique carotane-type “5 + 7” bicycle skeleton. Tang and coworkers reconstituted the biosynthesis of aspterric acid and demonstrated that sesquiterpene cyclase AstA and two cytochrome P450s (AstB and AstC) jointly catalyzed the production of aspterric acid (Yan et al. 2018). Thus, as outlined in Scheme 1B, the starter compound FPP was converted into carotane-type key intermediate (-)-daucane catalyzed by terpene cyclase Pb_CarA. The intermediate is converted into structurally determined α-epoxy carboxylate intermediate iii through CarB-catalyzed multi-step oxidation, in which the epoxy ring might be cleaved to afford trans-1,2-diol intermediate iv. The existence of the intermediate ii is supported by the isolation and characterization of 4 from the strain P. brasilianum. The intermediate iv might undergo C15-hydroxylation to generate intermediate v, which might be catalyzed by cytochrome P450 CarC. The intermediate iii would then be transformed into the final product aspterric acid, in which the C9–C15 β-ether oxygen moiety was formed. In addition, the biosynthesizing pathway of compound 3 might undergo dihydroxylation of the C11–C12 ethylene bond. The step of oxidative modification might be catalyzed by a separated oxygenase in Penicllium or co-culture pairing strain Armillaria.

Cytotoxic activities of induced representative metabolites

These compounds (1–6) were evaluated for their cytotoxic activities against MCF-7, A549, and HepG2 cell lines in vitro with cisplatin as the positive control. All six compounds have no obvious inhibition activities against three cell lines (Table S6, supporting information).

Discussion

In this report, we have documented the endeavor to unearth the chemical repertoire of co-culture system including P. brasilianum and A. mellea species. Interestingly, this has led to the discovery of new compounds that were not detected in monoculture. Since the study of the fungal-fungal co-cultivation of Acremonium sp. Tbp-5 and Mycogone rosea DSM 12973 two decades ago (Degenkolb et al. 2002), the co-culture approach has accelerated the application in mining SMs from fungal species, and numerous compounds exhibiting unique architectures or potent bioactivities have been identified (Goers et al. 2014; Knowles et al. 2022). In addition, with genome sequences becoming accessible due to decreasing costs, genome-guided strategies are now accessible and have revolutionized the discovery of bioactive metabolites (Zhang et al. 2023b, 2022b). Previous studies demonstrated that the transcriptional induction of BGCs is controlled by environmental or interspecies motivation (Knowles et al. 2022; Zhang et al. 2022a). Therefore, considering the biosynthetic potential of the fungus P. brasilianum, an OSMAC and a co-culture approach were performed to activate the production of novel SMs, respectively. However, the OSMAC strategy did not exhibit the capability of the strain P. brasilianum model (Fig. S1, supporting information). In contrast, during co-culturing with Armillaria, the metabolic spectrum of the strain P. brasilianum has been markedly broadened and new sesquiterpene compounds were generated (Figs. 1 and 2). Interestingly, previous studies conducted by Ding and colleagues utilized a liquid co-culture system of Armillaria sp. and Epicoccum sp., and new protoilludane-derived epicoterpenes and armilliphatics A synthesized by Armillaria were isolated (Li et al. 2020). But based on our study, nevertheless, the paring fungus Armillaria mainly acted as an inducer and most metabolites accumulated in the co-culture were produced by the fungus P. brasilianum. This result also highlighted the complexity of the metabolomic process of fungal interspecies interactions, suggesting the different response mechanisms of fungal chemical defenses or inductions (Knowles et al. 2022). Taken together, our present study demonstrates that this approach is a promising strategy to broaden the metabolic patterns of even for well-studied Penicillium strains.

To the best of our knowledge, this is the first study that two different categories of sesquiterpenes were identified from the strain P. brasilianum. Previous studies have already manifested the metabolic potential of P. brasilianum species, and a diverse assortment of bioactive metabolites have been isolated (Bazioli et al. 2017). A more detailed investigation of P. brasilianum species and metabolites produced has already been conducted, including the isolate P. brasilianum LaBioMMI 024 (producing preaustinoids, verruculogens), the isolate JV-379 (producing brasiliamides), P. brasilianum Batista and Penicillium sp. MG-11 (producing austin, penicillic acid), P. brasilianum NBRC 6234 (producing paraherquonin), the isolate LaBioMMi 136 (producing cyclodepsipeptides JBIR 113), and the isolate FKI-3368 (producing spirohexaline, viridicatumtoxin), for detailed information reference review paper by Bazioli et al. (2017). However, for the strain P. brasilianum, no sesquiterpenoids have been reported so far. In our report, conspicuously, the arpenibisabolanes (1–2) and arpenicarotanes (3–4) were identified. Arpenibisabolanes 1 and 2 were biosynthesized by bisabolene synthase Pb_BisA, which exhibited higher identity to UbiA-type α-bisabolol synthase (BibS) from Fusarium sp., Stachybotrys sp., and A. absinthium (Luo et al. 2022; Muangphrom et al. 2016). Noteworthy, Pb_BisA from P. brasilianum was distributed in the BibS branch (Fig. 5A). This group of sesquiterpene synthases constitutes a new subfamily and only few fungal-derived BibSs have been characterized (Luo et al. 2022). For the metabolites of carotane-type sesquiterpenes, aspterric acid was first discovered from A. terreus IFO-6123 (Tsuda et al. 1978) and the biosynthetic pathway was deciphered using yeast-based heterologous expression platform by Tang and colleagues (Yan et al. 2018). Interestingly, uncommon compounds 1 and 3 exhibited complexified oxidations, e.g., furan ring formation in 1, and C11–C12 dihydroxylations in 3; these biosynthetic processes might be catalyzed by oxygenases from paring Armillaria species. New metabolites induced by co-culture including P. brasilianum and A. mellea revealed a complex molecular mechanism involving interspecies induction or biotransformation.

In the course of our studies, we could not detect the cytotoxic activities of isolated sesquiterpenoids on carcinogenic cell lines. Nevertheless, compound 6 was reported as potent herbicidal agent (Tsuda et al. 1978; Yan et al. 2018). In addition, for the metabolites of bisabolene sesquiterpene derivatives, (Z)-12-acetoxybisabol-1-one and 12-acetoxybisabolen-1-ol from Trichoderma asperellum EN-764 exhibiting inhibitory activities (4–16 µg/mL) against aquatic pathogens including Vibrio alginolyticus and V. harveyi were reported (Li et al. 2023a). This implies that the two classes of sesquiterpenoids might function as herbicides or possess antibacterial activities. With the strain and genomic information, our understanding of the biosynthesis of intriguing sesquiterpenoids could be promoted by employing heterologous reconstitution or metabolic engineering strategies.

To summarize, the co-cultivation system of P. brasilianum and A. mellea was conducted to induce new SMs production and compound discovery. Four novel sesquiterpenoids (1–4) and one new polyketide 5 were identified from their co-cultivation. Among these, compound 1 possessed a 6/5-fused bicyclic ring scaffold, representing the first example of a 6/5-fused bicyclic bisabolene. These findings will enrich the chemical diversity of sesquiterpenoids and stimulate the interest from academic researchers. Consequently, this study might serve as an example that co-culturing is an attractive approach for broadening the chemical profile of microbes.

Supplementary Information

Below is the link to the electronic supplementary material.Supplementary file1 (PDF 4862 KB)

Author contribution

T.Z., L.X.Z., and L.Y. conceived the study. T.Z. obtained initial funding and supervised experiments. T.Z. and X.R. carried out experiments and bioinformatic analysis. L.H.Z. and W.H. elucidated the structures of the compounds. Z.G., J.B., and H.L. conducted the carcinoma cell assay. T.Z. and L.H.Z. wrote the draft manuscript. T.Z. edited the draft manuscript. All authors reviewed the final version of the manuscript.

Funding

This work was financially supported by the National Natural Science Foundation of China (No. 31872617), the CAMS Innovation Fund for Medical Sciences (CIFMS) (2021-I2M-1–055, 2019-I2M-1–005), the Central Level, Scientific Research Institutes for Basic R & D Fund Business (3332018097), and National Microbial Resource Center, (No. NMRC-2024-3).

Data availability

The raw read sequences of the biosynthetic gene clusters (Pb_car and Pb_bis) genes in this study are publicly available in the NCBI database (PP700700 and PP700701). The authors declare that the data supporting the findings of this study are available within the paper and its Supplementary Information files. Should any raw data files be needed in another format, they are available from the corresponding author upon reasonable request.

Declarations

Conflict of interest

The authors declare no competing interests.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Xiaoting Rong and Lihua Zhang contributed equally to this work.
==== Refs
References

Ariantari NP Daletos G Mandi A Kurtan T Muller WEG Lin W Ancheeva E Proksch P Expanding the chemical diversity of an endophytic fungus Bulgaria inquinans, an ascomycete associated with mistletoe, through an OSMAC approach RSC Adv 2019 9 43 25119 25132 10.1039/c9ra03678d 35528664
Ariantari NP, Daletos G, Mandi A, Kurtan T, Muller WEG, Lin W, Ancheeva E, Proksch P (2019) Expanding the chemical diversity of an endophytic fungus Bulgaria inquinans, an ascomycete associated with mistletoe, through an OSMAC approach. RSC Adv 9(43):25119–25132. 10.1039/c9ra03678d35528664 10.1039/c9ra03678d
Bateman A, Martin MJ, Orchard S, Magrane M, Ahmad S, Alpi E, Bowler-Barnett EH, Britto R, Cukura A, Denny P, Dogan T, Ebenezer T, Fan J, Garmiri P, Gonzales LJD, Hatton-Ellis E, Hussein A, Ignatchenko A, Insana G, Ishtiaq R, Joshi V, Jyothi D, Kandasaamy S, Lock A, Luciani A, Lugaric M, Luo J, Lussi Y, MacDougall A, Madeira F, Mahmoudy M, Mishra A, Moulang K, Nightingale A, Pundir S, Qi GY, Raj S, Raposo P, Rice DL, Saidi R, Santos R, Speretta E, Stephenson J, Totoo P, Turner E, Tyagi N, Vasudev P, Warner K, Watkins X, Zellner H, Bridge AJ, Aimo L, Argoud-Puy GL, Auchincloss AH, Axelsen KB, Bansal P, Baratin D, Neto TMB, Blatter MC, Bolleman JT, Boutet E, Breuza L, Gil BC, Casals-Casas C, Echioukh KC, Coudert E, Cuche B, de Castro E, Estreicher A, Famiglietti ML, Feuermann M, Gasteiger E, Gaudet P, Gehant S, Gerritsen V, Gos A, Gruaz N, Hulo C, Hyka-Nouspikel N, Jungo F, Kerhornou A, Le Mercier P, Lieberherr D, Masson P, Morgat A, Muthukrishnan V, Paesano S, Pedruzzi I, Pilbout S, Pourcel L, Poux S, Pozzato M, Pruess M, Redaschi N, Rivoire C, Sigrist CJA, Sonesson K, Arighi CN, Armin-ski L, Chen CM, Chen YX, Huang HZ, Laiho K, McGarvey P, Natale DA, Ross K, Vinayaka CR, Wang QH, Wang YQ, Zhang J, Bye-A-Jee H, Zaru R, Sundaram S, Wu CH, Consortium U UniProt: the universal protein knowledgebase in 2023 Nucleic Acids Res 2023 51 D1 D523 D531 10.1093/nar/gkac1052 36408920
Bateman A, Martin MJ, Orchard S, Magrane M, Ahmad S, Alpi E, Bowler-Barnett EH, Britto R, Cukura A, Denny P, Dogan T, Ebenezer T, Fan J, Garmiri P, Gonzales LJD, Hatton-Ellis E, Hussein A, Ignatchenko A, Insana G, Ishtiaq R, Joshi V, Jyothi D, Kandasaamy S, Lock A, Luciani A, Lugaric M, Luo J, Lussi Y, MacDougall A, Madeira F, Mahmoudy M, Mishra A, Moulang K, Nightingale A, Pundir S, Qi GY, Raj S, Raposo P, Rice DL, Saidi R, Santos R, Speretta E, Stephenson J, Totoo P, Turner E, Tyagi N, Vasudev P, Warner K, Watkins X, Zellner H, Bridge AJ, Aimo L, Argoud-Puy GL, Auchincloss AH, Axelsen KB, Bansal P, Baratin D, Neto TMB, Blatter MC, Bolleman JT, Boutet E, Breuza L, Gil BC, Casals-Casas C, Echioukh KC, Coudert E, Cuche B, de Castro E, Estreicher A, Famiglietti ML, Feuermann M, Gasteiger E, Gaudet P, Gehant S, Gerritsen V, Gos A, Gruaz N, Hulo C, Hyka-Nouspikel N, Jungo F, Kerhornou A, Le Mercier P, Lieberherr D, Masson P, Morgat A, Muthukrishnan V, Paesano S, Pedruzzi I, Pilbout S, Pourcel L, Poux S, Pozzato M, Pruess M, Redaschi N, Rivoire C, Sigrist CJA, Sonesson K, Arighi CN, Armin-ski L, Chen CM, Chen YX, Huang HZ, Laiho K, McGarvey P, Natale DA, Ross K, Vinayaka CR, Wang QH, Wang YQ, Zhang J, Bye-A-Jee H, Zaru R, Sundaram S, Wu CH, Consortium U (2023) UniProt: the universal protein knowledgebase in 2023. Nucleic Acids Res 51(D1):D523–D531. 10.1093/nar/gkac105236408920 10.1093/nar/gkac1052
Baumgartner K Coetzee MP Hoffmeister D Secrets of the subterranean pathosystem of Armillaria Mol Plant Pathol 2011 12 6 515 534 10.1111/j.1364-3703.2010.00693.x 21722292
Baumgartner K, Coetzee MP, Hoffmeister D (2011) Secrets of the subterranean pathosystem of Armillaria. Mol Plant Pathol 12(6):515–534. 10.1111/j.1364-3703.2010.00693.x21722292 10.1111/j.1364-3703.2010.00693.x
Bazioli JM Amaral LDS Fill TP Rodrigues-Filho E Insights into Penicillium brasilianum secondary metabolism and its biotechnological potential Molecules 2017 22 6 858 10.3390/molecules22060858 28632186
Bazioli JM, Amaral LDS, Fill TP, Rodrigues-Filho E (2017) Insights into Penicillium brasilianum secondary metabolism and its biotechnological potential. Molecules 22(6):858. 10.3390/molecules2206085828632186 10.3390/molecules22060858
Bertrand S Bohni N Schnee S Schumpp O Gindro K Wolfender JL Metabolite induction via microorganism co-culture: a potential way to enhance chemical diversity for drug discovery Biotechnol Adv 2014 32 6 1180 1204 10.1016/j.biotechadv.2014.03.001 24651031
Bertrand S, Bohni N, Schnee S, Schumpp O, Gindro K, Wolfender JL (2014) Metabolite induction via microorganism co-culture: a potential way to enhance chemical diversity for drug discovery. Biotechnol Adv 32(6):1180–1204. 10.1016/j.biotechadv.2014.03.00124651031 10.1016/j.biotechadv.2014.03.001
Bills GF Gloer JB Biologically active secondary metabolites from the fungi Microbiol Spectr 2016 4 6 10 128 10.1128/microbiolspec.FUNK-0009-2016
Bills GF, Gloer JB (2016) Biologically active secondary metabolites from the fungi. Microbiol Spectr 4(6):10–128. 10.1128/microbiolspec.FUNK-0009-201610.1128/microbiolspec.FUNK-0009-2016
Blin K Pascal Andreu V de Los Santos ELC Del Carratore F Lee SY Medema MH Weber T The antiSMASH database version 2: a comprehensive resource on secondary metabolite biosynthetic gene clusters Nucleic Acids Res 2019 47 D1 D625 D630 10.1093/nar/gky1060 30395294
Blin K, Pascal Andreu V, de Los Santos ELC, Del Carratore F, Lee SY, Medema MH, Weber T (2019) The antiSMASH database version 2: a comprehensive resource on secondary metabolite biosynthetic gene clusters. Nucleic Acids Res 47(D1):D625–D630. 10.1093/nar/gky106030395294 10.1093/nar/gky1060
Bode HB Bethe B Höfs R Zeeck A Big effects from small changes: possible ways to explore nature’s chemical diversity ChemBioChem 2002 3 7 619 627 10.1002/1439-7633(20020703)3:7<619::AID-CBIC619>3.0.CO;2-9 12324995
Bode HB, Bethe B, Höfs R, Zeeck A (2002) Big effects from small changes: possible ways to explore nature’s chemical diversity. ChemBioChem 3(7):619–627. 10.1002/1439-7633(20020703)3:7%3c619::AID-CBIC619%3e3.0.CO;2-912324995 10.1002/1439-7633(20020703)3:7<619::AID-CBIC619>3.0.CO;2-9
Bouws H Wattenberg A Zorn H Fungal secretomes–nature’s toolbox for white biotechnology Appl Microbiol Biotechnol 2008 80 3 381 388 10.1007/s00253-008-1572-5 18636256
Bouws H, Wattenberg A, Zorn H (2008) Fungal secretomes–nature’s toolbox for white biotechnology. Appl Microbiol Biotechnol 80(3):381–388. 10.1007/s00253-008-1572-518636256 10.1007/s00253-008-1572-5
Brase S Encinas A Keck J Nising CF Chemistry and biology of mycotoxins and related fungal metabolites Chem Rev 2009 109 9 3903 3990 10.1021/cr050001f 19534495
Brase S, Encinas A, Keck J, Nising CF (2009) Chemistry and biology of mycotoxins and related fungal metabolites. Chem Rev 109(9):3903–3990. 10.1021/cr050001f19534495 10.1021/cr050001f
Cichewicz RH Epigenome manipulation as a pathway to new natural product scaffolds and their congeners Nat Prod Rep 2010 27 1 11 22 10.1039/b920860g 20024091
Cichewicz RH (2010) Epigenome manipulation as a pathway to new natural product scaffolds and their congeners. Nat Prod Rep 27(1):11–22. 10.1039/b920860g20024091 10.1039/b920860g
Degenkolb T Heinze S Schlegel B Strobel G Grafe U Formation of new lipoaminopeptides, acremostatins A, B, and C, by co-cultivation of Acremonium sp. Tbp-5 and Mycogone rosea DSM 12973 Biosci Biotechnol Biochem 2002 66 4 883 886 10.1271/bbb.66.883 12036069
Degenkolb T, Heinze S, Schlegel B, Strobel G, Grafe U (2002) Formation of new lipoaminopeptides, acremostatins A, B, and C, by co-cultivation of Acremonium sp. Tbp-5 and Mycogone rosea DSM 12973. Biosci Biotechnol Biochem 66(4):883–886. 10.1271/bbb.66.88312036069 10.1271/bbb.66.883
Elhamouly NA Hewedy OA Zaitoon A Miraples A Elshorbagy OT Hussien S El-Tahan A Peng D The hidden power of secondary metabolites in plant-fungi interactions and sustainable phytoremediation Front Plant Sci 2022 13 1044896 10.3389/fpls.2022.1044896 36578344
Elhamouly NA, Hewedy OA, Zaitoon A, Miraples A, Elshorbagy OT, Hussien S, El-Tahan A, Peng D (2022) The hidden power of secondary metabolites in plant-fungi interactions and sustainable phytoremediation. Front Plant Sci 13:1044896. 10.3389/fpls.2022.104489636578344 10.3389/fpls.2022.1044896
Fradj N de Montigny N Merindol N Awwad F Boumghar Y Germain H Desgagne-Penix I A first insight into north American plant pathogenic fungi Armillaria sinapina transcriptome Biology (Basel) 2020 9 7 153 10.3390/biology9070153 32635577
Fradj N, de Montigny N, Merindol N, Awwad F, Boumghar Y, Germain H, Desgagne-Penix I (2020) A first insight into north American plant pathogenic fungi Armillaria sinapina transcriptome. Biology (Basel) 9(7):153. 10.3390/biology907015332635577 10.3390/biology9070153
Ghoul M Mitri S The ecology and evolution of microbial competition Trends Microbiol 2016 24 10 833 845 10.1016/j.tim.2016.06.011 27546832
Ghoul M, Mitri S (2016) The ecology and evolution of microbial competition. Trends Microbiol 24(10):833–845. 10.1016/j.tim.2016.06.01127546832 10.1016/j.tim.2016.06.011
Goers L Freemont P Polizzi KM Co-culture systems and technologies: taking synthetic biology to the next level J R Soc Interface 2014 11 96 20140065 10.1098/rsif.2014.0065 24829281
Goers L, Freemont P, Polizzi KM (2014) Co-culture systems and technologies: taking synthetic biology to the next level. J R Soc Interface 11(96):20140065. 10.1098/rsif.2014.006524829281 10.1098/rsif.2014.0065
Jones CG Moniodis J Zulak KG Scaffidi A Plummer JA Ghisalberti EL Barbour EL Bohlmann J Sandalwood fragrance biosynthesis involves sesquiterpene synthases of both the terpene synthase (TPS)-a and TPS-b subfamilies, including santalene synthases J Biol Chem 2011 286 20 17445 17454 10.1074/jbc.M111.231787 21454632
Jones CG, Moniodis J, Zulak KG, Scaffidi A, Plummer JA, Ghisalberti EL, Barbour EL, Bohlmann J (2011) Sandalwood fragrance biosynthesis involves sesquiterpene synthases of both the terpene synthase (TPS)-a and TPS-b subfamilies, including santalene synthases. J Biol Chem 286(20):17445–17454. 10.1074/jbc.M111.23178721454632 10.1074/jbc.M111.231787
Keller NP Fungal secondary metabolism: regulation, function and drug discovery Nat Rev Microbiol 2019 17 3 167 180 10.1038/s41579-018-0121-1 30531948
Keller NP (2019) Fungal secondary metabolism: regulation, function and drug discovery. Nat Rev Microbiol 17(3):167–180. 10.1038/s41579-018-0121-130531948 10.1038/s41579-018-0121-1
Kim JH Lee N Hwang S Kim W Lee Y Cho S Palsson BO Cho BK Discovery of novel secondary metabolites encoded in actinomycete genomes through coculture J Ind Microbiol Biotechnol 2021 48 3–4 kuaa001 10.1093/jimb/kuaa001 33825906
Kim JH, Lee N, Hwang S, Kim W, Lee Y, Cho S, Palsson BO, Cho BK (2021) Discovery of novel secondary metabolites encoded in actinomycete genomes through coculture. J Ind Microbiol Biotechnol 48(3–4):kuaa001. 10.1093/jimb/kuaa00133825906 10.1093/jimb/kuaa001
Knowles SL Raja HA Roberts CD Oberlies NH Fungal-fungal co-culture: a primer for generating chemical diversity Nat Prod Rep 2022 39 8 1557 1573 10.1039/d1np00070e 35137758
Knowles SL, Raja HA, Roberts CD, Oberlies NH (2022) Fungal-fungal co-culture: a primer for generating chemical diversity. Nat Prod Rep 39(8):1557–1573. 10.1039/d1np00070e35137758 10.1039/d1np00070e
Kozlovsky AG Kochkina GA Zhelifonova VP Antipova ТV Ivanushkina NE Ozerskaya SM Secondary metabolites of the genus Penicillium from undisturbed and anthropogenically altered Antarctic habitats Folia Microbiol (Praha) 2020 65 1 95 102 10.1007/s12223-019-00708-0 30982204
Kozlovsky AG, Kochkina GA, Zhelifonova VP, Antipova ТV, Ivanushkina NE, Ozerskaya SM (2020) Secondary metabolites of the genus Penicillium from undisturbed and anthropogenically altered Antarctic habitats. Folia Microbiol (Praha) 65(1):95–102. 10.1007/s12223-019-00708-030982204 10.1007/s12223-019-00708-0
Lewis EA Adamek TL Vining LC White RL Metabolites of a blocked chloramphenicol producer J Nat Prod 2003 66 1 62 66 10.1021/np020306e 12542347
Lewis EA, Adamek TL, Vining LC, White RL (2003) Metabolites of a blocked chloramphenicol producer. J Nat Prod 66(1):62–66. 10.1021/np020306e12542347 10.1021/np020306e
Li H Tang L Liu T Yang R Yang Y Zhou H Ding ZT Protoilludane-type sesquiterpenoids from Armillaria sp. by co-culture with the endophytic fungus Epicoccum sp. associated with Gastrodia elata Bioorg Chem 2020 95 103503 10.1016/j.bioorg.2019.103503 31855825
Li H, Tang L, Liu T, Yang R, Yang Y, Zhou H, Ding ZT (2020) Protoilludane-type sesquiterpenoids from Armillaria sp. by co-culture with the endophytic fungus Epicoccum sp. associated with Gastrodia elata. Bioorg Chem 95:103503. 10.1016/j.bioorg.2019.10350331855825 10.1016/j.bioorg.2019.103503
Li H Li X Ying Z Li Y Wang B Bisabolane sesquiterpene and cyclopentene derivatives from the marine algal-derived endophytic fungus Trichoderma asperellum EN-764 Phytochemistry 2023 210 113644 10.1016/j.phytochem.2023.113644 36935049
Li H, Li X, Ying Z, Li Y, Wang B (2023) Bisabolane sesquiterpene and cyclopentene derivatives from the marine algal-derived endophytic fungus Trichoderma asperellum EN-764. Phytochemistry 210:113644. 10.1016/j.phytochem.2023.11364436935049 10.1016/j.phytochem.2023.113644
Li Y Feng J Li J Li X Wang H Yu L Li J Zhang T Diversity and functional research of sesquiterpene synthases in fungus Armillaria mellea CPCC 400891 Biotic Resources 2023 45 4 355 364 10.14188/j.ajsh.2023.04.007
Li Y, Feng J, Li J, Li X, Wang H, Yu L, Li J, Zhang T (2023) Diversity and functional research of sesquiterpene synthases in fungus Armillaria mellea CPCC 400891. Biotic Resources 45(4):355–364. 10.14188/j.ajsh.2023.04.00710.14188/j.ajsh.2023.04.007
Lin HC Chooi YH Dhingra S Xu W Calvo AM Tang Y The fumagillin biosynthetic gene cluster in Aspergillus fumigatus encodes a cryptic terpene cyclase involved in the formation of beta-trans-bergamotene J Am Chem Soc 2013 135 12 4616 4619 10.1021/ja312503y 23488861
Lin HC, Chooi YH, Dhingra S, Xu W, Calvo AM, Tang Y (2013) The fumagillin biosynthetic gene cluster in Aspergillus fumigatus encodes a cryptic terpene cyclase involved in the formation of beta-trans-bergamotene. J Am Chem Soc 135(12):4616–4619. 10.1021/ja312503y23488861 10.1021/ja312503y
Luo R Liu B Xie Y Li Z Huang W Yuan J He G Chen Y Pan Q Liu Y Tang J Wu G Zhang H Shi Y Liu Y Yu C Wang B Lu Y Han C Cheung D Yiu S Peng S Zhu X Liu G Liao X Li Y Yang H Wang J Lam T Wang J (2015) SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler Gigascience 2012 1 1 18 10.1186/2047-217X-1-18 23587118
Luo R, Liu B, Xie Y, Li Z, Huang W, Yuan J, He G, Chen Y, Pan Q, Liu Y, Tang J, Wu G, Zhang H, Shi Y, Liu Y, Yu C, Wang B, Lu Y, Han C, Cheung D, Yiu S, Peng S, Zhu X, Liu G, Liao X, Li Y, Yang H, Wang J, Lam T, Wang J (2012) SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler. Gigascience 1(1):18. 10.1186/2047-217X-1-1823587118 10.1186/2047-217X-1-18
Luo P Lv J Xie Y Xiao L Qin S Chen G Luo X Hu D Gao H Discovery and characterization of a novel sub-group of UbiA-type terpene cyclases with a distinct motif I Org Chem Front 2022 9 11 3057 3060 10.1039/D2QO00408A
Luo P, Lv J, Xie Y, Xiao L, Qin S, Chen G, Luo X, Hu D, Gao H (2022) Discovery and characterization of a novel sub-group of UbiA-type terpene cyclases with a distinct motif I. Org Chem Front 9(11):3057–3060. 10.1039/D2QO00408A10.1039/D2QO00408A
Lyu HN Zhang J Zhou S Liu HW Zhuang WY Li SM Yin WB Heterologous expression of a single fungal HR-PKS leads to the formation of diverse 2-alkenyl-tetrahydropyrans in model fungi Org Biomol Chem 2021 19 38 8377 8383 10.1039/d1ob01501j 34528986
Lyu HN, Zhang J, Zhou S, Liu HW, Zhuang WY, Li SM, Yin WB (2021) Heterologous expression of a single fungal HR-PKS leads to the formation of diverse 2-alkenyl-tetrahydropyrans in model fungi. Org Biomol Chem 19(38):8377–8383. 10.1039/d1ob01501j34528986 10.1039/d1ob01501j
Malit JJL Leung HYC Qian P Targeted large-scale genome mining and candidate prioritization for natural product discovery Mar Drugs 2022 20 6 398 10.3390/md20060398 35736201
Malit JJL, Leung HYC, Qian P (2022) Targeted large-scale genome mining and candidate prioritization for natural product discovery. Mar Drugs 20(6):398. 10.3390/md2006039835736201 10.3390/md20060398
Mao X Xu W Li D Yin WB Chooi YH Li Y Tang Y Hu Y Epigenetic genome mining of an endophytic fungus leads to the pleiotropic biosynthesis of natural products Angew Chem Int Ed Engl 2015 54 26 7592 7596 10.1002/anie.201502452 26013262
Mao X, Xu W, Li D, Yin WB, Chooi YH, Li Y, Tang Y, Hu Y (2015) Epigenetic genome mining of an endophytic fungus leads to the pleiotropic biosynthesis of natural products. Angew Chem Int Ed Engl 54(26):7592–7596. 10.1002/anie.20150245226013262 10.1002/anie.201502452
Medema MH de Rond T Moore BS Mining genomes to illuminate the specialized chemistry of life Nat Rev Genet 2021 22 9 553 571 10.1038/s41576-021-00363-7 34083778
Medema MH, de Rond T, Moore BS (2021) Mining genomes to illuminate the specialized chemistry of life. Nat Rev Genet 22(9):553–571. 10.1038/s41576-021-00363-734083778 10.1038/s41576-021-00363-7
Muangphrom P Seki H Suzuki M Komori A Nishiwaki M Mikawa R Fukushima EO Muranaka T Functional analysis of amorpha-4,11-diene synthase (ADS) homologs from non-artemisinin-producing Artemisia species: the discovery of novel koidzumiol and (+)-alpha-bisabolol synthases Plant Cell Physiol 2016 57 8 1678 1688 10.1093/pcp/pcw094 27273626
Muangphrom P, Seki H, Suzuki M, Komori A, Nishiwaki M, Mikawa R, Fukushima EO, Muranaka T (2016) Functional analysis of amorpha-4,11-diene synthase (ADS) homologs from non-artemisinin-producing Artemisia species: the discovery of novel koidzumiol and (+)-alpha-bisabolol synthases. Plant Cell Physiol 57(8):1678–1688. 10.1093/pcp/pcw09427273626 10.1093/pcp/pcw094
Ohtani K Fujioka S Kawano T Shimada A Kimura Y Nematicidal activities of 4-hydroxyphenylacetic acid and oidiolactone D produced by the fungus Oidiodendron sp Z Naturforsch C J Biosci 2011 66 1–2 31 34 10.1515/znc-2011-1-205 21476434
Ohtani K, Fujioka S, Kawano T, Shimada A, Kimura Y (2011) Nematicidal activities of 4-hydroxyphenylacetic acid and oidiolactone D produced by the fungus Oidiodendron sp. Z Naturforsch C J Biosci 66(1–2):31–34. 10.1515/znc-2011-1-20521476434 10.1515/znc-2011-1-205
Oikawa H Heterologous production of fungal natural products: reconstitution of biosynthetic gene clusters in model host Aspergillus oryzae Proc Jpn Acad Ser B Phys Biol Sci 2020 96 9 420 430 10.2183/pjab.96.031 33177296
Oikawa H (2020) Heterologous production of fungal natural products: reconstitution of biosynthetic gene clusters in model host Aspergillus oryzae. Proc Jpn Acad Ser B Phys Biol Sci 96(9):420–430. 10.2183/pjab.96.03133177296 10.2183/pjab.96.031
Pan R Bai X Chen J Zhang H Wang H Exploring structural diversity of microbe secondary metabolites using OSMAC strategy: a literature review Front Microbiol 2019 10 294 10.3389/fmicb.2019.00294 30863377
Pan R, Bai X, Chen J, Zhang H, Wang H (2019) Exploring structural diversity of microbe secondary metabolites using OSMAC strategy: a literature review. Front Microbiol 10:294. 10.3389/fmicb.2019.0029430863377 10.3389/fmicb.2019.00294
Pan G Li Y Che X Tian D Han W Wang Z Zhao Y Ren S Xu Y Hao G Guo M Xiao N Kong F New thio-compounds and monoterpenes with anti-inflammatory activities from the fungus Aspergillus sp. CYH26 Front Microbiol 2021 12 668938 10.3389/fmicb.2021.668938 33841388
Pan G, Li Y, Che X, Tian D, Han W, Wang Z, Zhao Y, Ren S, Xu Y, Hao G, Guo M, Xiao N, Kong F (2021) New thio-compounds and monoterpenes with anti-inflammatory activities from the fungus Aspergillus sp. CYH26. Front Microbiol 12:668938. 10.3389/fmicb.2021.66893833841388 10.3389/fmicb.2021.668938
Park HB Park JS Lee SI Shin B Oh DC Kwon HC Gordonic acid, a polyketide glycoside derived from bacterial coculture of Streptomyces and Gordonia species J Nat Prod 2017 80 9 2542 2546 10.1021/acs.jnatprod.7b00293 28845982
Park HB, Park JS, Lee SI, Shin B, Oh DC, Kwon HC (2017) Gordonic acid, a polyketide glycoside derived from bacterial coculture of Streptomyces and Gordonia species. J Nat Prod 80(9):2542–2546. 10.1021/acs.jnatprod.7b0029328845982 10.1021/acs.jnatprod.7b00293
Ralph J Quideau S Grabber JH Hatfield RD Identification and synthesis of new ferulic acid dehydrodimers present in grass cell walls J Chem Soc, Perkin Trans 1994 1 23 3485 3498 10.1039/P19940003485
Ralph J, Quideau S, Grabber JH, Hatfield RD (1994) Identification and synthesis of new ferulic acid dehydrodimers present in grass cell walls. J Chem Soc, Perkin Trans 1(23):3485–3498. 10.1039/P1994000348510.1039/P19940003485
Ro DK Ehlting J Keeling CI Lin R Mattheus N Bohlmann J Microarray expression profiling and functional characterization of AtTPS genes: duplicated Arabidopsis thaliana sesquiterpene synthase genes At4g13280 and At4g13300 encode root-specific and wound-inducible (Z)-gamma-bisabolene synthases Arch Biochem Biophys 2006 448 1–2 104 116 10.1016/j.abb.2005.09.019 16297850
Ro DK, Ehlting J, Keeling CI, Lin R, Mattheus N, Bohlmann J (2006) Microarray expression profiling and functional characterization of AtTPS genes: duplicated Arabidopsis thaliana sesquiterpene synthase genes At4g13280 and At4g13300 encode root-specific and wound-inducible (Z)-gamma-bisabolene synthases. Arch Biochem Biophys 448(1–2):104–116. 10.1016/j.abb.2005.09.01916297850 10.1016/j.abb.2005.09.019
Rong X Guo Z He W Cai G Gong K Wang L Yu L Zhang T Gao K Secondary metabolites exhibiting antitumor bioactivities from the fungus Stachybotrys sp CPCC 401591 Mycosystema 2023 42 7 1611 1621 10.13346/j.mycosystema.220342
Rong X, Guo Z, He W, Cai G, Gong K, Wang L, Yu L, Zhang T, Gao K (2023a) Secondary metabolites exhibiting antitumor bioactivities from the fungus Stachybotrys sp CPCC 401591. Mycosystema 42(7):1611–1621. 10.13346/j.mycosystema.22034210.13346/j.mycosystema.220342
Rong X He W Guo Z Li X Wang L Gao K Yu L Zhang T Isolation and antitumor activity of phenylspirodrimane derivatives from the fungus Stachybotrys sp CPCC 401591 Nat Prod Res Dev 2023 35 8 1348 1356 10.16333/j.1001-6880.2023.8.007
Rong X, He W, Guo Z, Li X, Wang L, Gao K, Yu L, Zhang T (2023b) Isolation and antitumor activity of phenylspirodrimane derivatives from the fungus Stachybotrys sp CPCC 401591. Nat Prod Res Dev 35(8):1348–1356. 10.16333/j.1001-6880.2023.8.00710.16333/j.1001-6880.2023.8.007
Shen S Li W Wang J A novel and other bioactive secondary metabolites from a marine fungus Penicillium oxalicum 0312F1 Nat Prod Res 2013 27 24 2286 2291 10.1080/14786419.2013.827190 23962399
Shen S, Li W, Wang J (2013) A novel and other bioactive secondary metabolites from a marine fungus Penicillium oxalicum 0312F1. Nat Prod Res 27(24):2286–2291. 10.1080/14786419.2013.82719023962399 10.1080/14786419.2013.827190
Sugiyama R Nakatani T Nishimura S Takenaka K Ozaki T Asamizu S Onaka H Kakeya H Chemical interactions of cryptic actinomycete metabolite 5-alkyl-1,2,3,4-tetrahydroquinolines through aggregate formation Angew Chem Int Ed Engl 2019 58 38 13486 13491 10.1002/anie.201905970 31389661
Sugiyama R, Nakatani T, Nishimura S, Takenaka K, Ozaki T, Asamizu S, Onaka H, Kakeya H (2019) Chemical interactions of cryptic actinomycete metabolite 5-alkyl-1,2,3,4-tetrahydroquinolines through aggregate formation. Angew Chem Int Ed Engl 58(38):13486–13491. 10.1002/anie.20190597031389661 10.1002/anie.201905970
Tsuda Y Kaneda M Tada A Nitta K Yamamoto Y Iitka Y Aspterric acid, a new sesquiterpenoid of carotane group, a metabolite from Aspergillus terreus IFO-6123 - X-ray crystal and molecular structure of its para-bromobenzoate J Chem Soc Chem Comm 1978 4 160 161 10.1039/c39780000160
Tsuda Y, Kaneda M, Tada A, Nitta K, Yamamoto Y, Iitka Y (1978) Aspterric acid, a new sesquiterpenoid of carotane group, a metabolite from Aspergillus terreus IFO-6123 - X-ray crystal and molecular structure of its para-bromobenzoate. J Chem Soc Chem Comm 4:160–161. 10.1039/c3978000016010.1039/c39780000160
Visagie CM Houbraken J Frisvad JC Hong SB Klaassen CH Perrone G Seifert KA Varga J Yaguchi T Samson RA Identification and nomenclature of the genus Penicillium Stud Mycol 2014 78 343 371 10.1016/j.simyco.2014.09.001 25505353
Visagie CM, Houbraken J, Frisvad JC, Hong SB, Klaassen CH, Perrone G, Seifert KA, Varga J, Yaguchi T, Samson RA (2014) Identification and nomenclature of the genus Penicillium. Stud Mycol 78:343–371. 10.1016/j.simyco.2014.09.00125505353 10.1016/j.simyco.2014.09.001
Wang W Zeng F Bie Q Dai C Chen C Tong Q Liu J Wang J Zhou Y Zhu H Zhang Y Cytochathiazines A-C: three merocytochalasans with a 2 H-1,4-thiazine functionality from coculture of Chaetomium globosum and Aspergillus flavipes Org Lett 2018 20 21 6817 6821 10.1021/acs.orglett.8b02942 30350674
Wang W, Zeng F, Bie Q, Dai C, Chen C, Tong Q, Liu J, Wang J, Zhou Y, Zhu H, Zhang Y (2018) Cytochathiazines A-C: three merocytochalasans with a 2 H-1,4-thiazine functionality from coculture of Chaetomium globosum and Aspergillus flavipes. Org Lett 20(21):6817–6821. 10.1021/acs.orglett.8b0294230350674 10.1021/acs.orglett.8b02942
Wang G Ran H Fan J Keller NP Liu Z Wu F Yin WB Fungal-fungal cocultivation leads to widespread secondary metabolite alteration requiring the partial loss-of-function VeA1 protein Sci Adv 2022 8 17 eabo6094 10.1126/sciadv.abo6094 35476435
Wang G, Ran H, Fan J, Keller NP, Liu Z, Wu F, Yin WB (2022) Fungal-fungal cocultivation leads to widespread secondary metabolite alteration requiring the partial loss-of-function VeA1 protein. Sci Adv 8(17):eabo6094. 10.1126/sciadv.abo609435476435 10.1126/sciadv.abo6094
Weiland-Brauer N Friends or foes-microbial interactions in nature Biology (Basel) 2021 10 6 496 10.3390/biology10060496 34199553
Weiland-Brauer N (2021) Friends or foes-microbial interactions in nature. Biology (Basel) 10(6):496. 10.3390/biology1006049634199553 10.3390/biology10060496
Xie X Mei W Zeng Y Lin H Lin H Zhuang L Dai H Hong K Cytotoxic constituents from marine actinomycete Streptomyces sp. 124092 Chemi J Chin Univ 2008 29 11 2183 2186
Xie X, Mei W, Zeng Y, Lin H, Lin H, Zhuang L, Dai H, Hong K (2008) Cytotoxic constituents from marine actinomycete Streptomyces sp. 124092. Chemi J Chin Univ 29(11):2183–2186
Xing CP Xie CL Xia JM Liu QM Lin WX Ye DZ Liu GM Yang XW Penigrisacids A-D, four new sesquiterpenes from the deep-sea-derived Penicillium griseofulvum Mar Drugs 2019 17 9 507 10.3390/md17090507 31470535
Xing CP, Xie CL, Xia JM, Liu QM, Lin WX, Ye DZ, Liu GM, Yang XW (2019) Penigrisacids A-D, four new sesquiterpenes from the deep-sea-derived Penicillium griseofulvum. Mar Drugs 17(9):507. 10.3390/md1709050731470535 10.3390/md17090507
Yan Y Liu Q Zang X Yuan S Bat-Erdene U Nguyen C Gan J Zhou J Jacobsen SE Tang Y Resistance-gene-directed discovery of a natural-product herbicide with a new mode of action Nature 2018 559 7714 415 418 10.1038/s41586-018-0319-4 29995859
Yan Y, Liu Q, Zang X, Yuan S, Bat-Erdene U, Nguyen C, Gan J, Zhou J, Jacobsen SE, Tang Y (2018) Resistance-gene-directed discovery of a natural-product herbicide with a new mode of action. Nature 559(7714):415–418. 10.1038/s41586-018-0319-429995859 10.1038/s41586-018-0319-4
Zang Y Gong Y Gong J Liu J Chen C Gu L Zhou Y Wang J Zhu H Zhang Y Fungal polyketides with three distinctive ring skeletons from the fungus Penicillium canescens uncovered by OSMAC and molecular networking strategies J Org Chem 2020 85 7 4973 4980 10.1021/acs.joc.0c00147 32118426
Zang Y, Gong Y, Gong J, Liu J, Chen C, Gu L, Zhou Y, Wang J, Zhu H, Zhang Y (2020) Fungal polyketides with three distinctive ring skeletons from the fungus Penicillium canescens uncovered by OSMAC and molecular networking strategies. J Org Chem 85(7):4973–4980. 10.1021/acs.joc.0c0014732118426 10.1021/acs.joc.0c00147
Zhang T Zhuo Y Jia XP Liu JT Gao H Song FH Liu M Zhang LX Cloning and characterization of the gene cluster required for beauvericin biosynthesis in Fusarium proliferatum Sci China Life Sci 2013 56 7 628 637 10.1007/s11427-013-4505-1 23832252
Zhang T, Zhuo Y, Jia XP, Liu JT, Gao H, Song FH, Liu M, Zhang LX (2013) Cloning and characterization of the gene cluster required for beauvericin biosynthesis in Fusarium proliferatum. Sci China Life Sci 56(7):628–637. 10.1007/s11427-013-4505-123832252 10.1007/s11427-013-4505-1
Zhang T Wan J Zhan Z Bai J Liu B Hu Y Activation of an unconventional meroterpenoid gene cluster in Neosartorya glabra leads to the production of new berkeleyacetals Acta Pharm Sin B 2018 8 3 478 487 10.1016/j.apsb.2017.12.005 29881687
Zhang T, Wan J, Zhan Z, Bai J, Liu B, Hu Y (2018) Activation of an unconventional meroterpenoid gene cluster in Neosartorya glabra leads to the production of new berkeleyacetals. Acta Pharm Sin B 8(3):478–487. 10.1016/j.apsb.2017.12.00529881687 10.1016/j.apsb.2017.12.005
Zhang T Cai G Rong X Wang Y Gong K Liu W Wang L Pang X Yu L A combination of genome mining with an OSMAC approach facilitates the discovery of and contributions to the biosynthesis of melleolides from the basidiomycete Armillaria tabescens J Agric Food Chem 2022 70 39 12430 12441 10.1021/acs.jafc.2c04079 36134616
Zhang T, Cai G, Rong X, Wang Y, Gong K, Liu W, Wang L, Pang X, Yu L (2022) A combination of genome mining with an OSMAC approach facilitates the discovery of and contributions to the biosynthesis of melleolides from the basidiomycete Armillaria tabescens. J Agric Food Chem 70(39):12430–12441. 10.1021/acs.jafc.2c0407936134616 10.1021/acs.jafc.2c04079
Zhang T Pang X Zhao J Guo Z He W Cai G Su J Cen S Yu L Discovery and activation of the cryptic cluster from Aspergillus sp. CPCC 400735 for asperphenalenone biosynthesis ACS Chem Biol 2022 17 6 1524 1533 10.1021/acschembio.2c00204 35616995
Zhang T, Pang X, Zhao J, Guo Z, He W, Cai G, Su J, Cen S, Yu L (2022) Discovery and activation of the cryptic cluster from Aspergillus sp. CPCC 400735 for asperphenalenone biosynthesis. ACS Chem Biol 17(6):1524–1533. 10.1021/acschembio.2c0020435616995 10.1021/acschembio.2c00204
Zhang T Feng J He W Rong X Lv H Li J Li X Wang H Wang L Zhang L Yu L Genomic and transcriptomic approaches provide a predictive framework for sesquiterpenes biosynthesis in Desarmillaria tabescens CPCC 401429 J Fungi (Basel) 2023 9 4 481 10.3390/jof9040481 37108935
Zhang T, Feng J, He W, Rong X, Lv H, Li J, Li X, Wang H, Wang L, Zhang L, Yu L (2023) Genomic and transcriptomic approaches provide a predictive framework for sesquiterpenes biosynthesis in Desarmillaria tabescens CPCC 401429. J Fungi (Basel) 9(4):481. 10.3390/jof904048137108935 10.3390/jof9040481
Zhang T Gu G Liu G Su J Zhan Z Zhao J Qian J Cai G Cen S Zhang D Yu L Late-stage cascade of oxidation reactions during the biosynthesis of oxalicine B in Penicillium oxalicum Acta Pharm Sin B 2023 13 1 256 270 10.1016/j.apsb.2022.09.008 36815048
Zhang T, Gu G, Liu G, Su J, Zhan Z, Zhao J, Qian J, Cai G, Cen S, Zhang D, Yu L (2023) Late-stage cascade of oxidation reactions during the biosynthesis of oxalicine B in Penicillium oxalicum. Acta Pharm Sin B 13(1):256–270. 10.1016/j.apsb.2022.09.00836815048 10.1016/j.apsb.2022.09.008
Zheng J Li Y Liu N Zhang J Liu S Tan H Multi-omics data reveal the effect of sodium butyrate on gene expression and protein modification in Streptomyces Genom Proteom Bioinform 2022 21 6 1149 1162 10.1016/j.gpb.2022.09.002
Zheng J, Li Y, Liu N, Zhang J, Liu S, Tan H (2022) Multi-omics data reveal the effect of sodium butyrate on gene expression and protein modification in Streptomyces. Genom Proteom Bioinform 21(6):1149–1162. 10.1016/j.gpb.2022.09.00210.1016/j.gpb.2022.09.002
Zhuang L Zhang H Utilizing cross-species co-cultures for discovery of novel natural products Curr Opin Biotechnol 2021 69 252 262 10.1016/j.copbio.2021.01.023 33647849
Zhuang L, Zhang H (2021) Utilizing cross-species co-cultures for discovery of novel natural products. Curr Opin Biotechnol 69:252–262. 10.1016/j.copbio.2021.01.02333647849 10.1016/j.copbio.2021.01.023
