Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Latest Articles
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • Topics
    • Applied and Environmental Science
    • Clinical Science and Epidemiology
    • Ecological and Evolutionary Science
    • Host-Microbe Biology
    • Molecular Biology and Physiology
    • Therapeutics and Prevention
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About mBio
    • Editor in Chief
    • Board of Editors
    • AAM Fellows
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
mBio
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Latest Articles
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • Topics
    • Applied and Environmental Science
    • Clinical Science and Epidemiology
    • Ecological and Evolutionary Science
    • Host-Microbe Biology
    • Molecular Biology and Physiology
    • Therapeutics and Prevention
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About mBio
    • Editor in Chief
    • Board of Editors
    • AAM Fellows
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
Research Article | Molecular Biology and Physiology

Monasone Naphthoquinone Biosynthesis and Resistance in Monascus Fungi

Mu Li, Lijing Kang, Xiaoli Ding, Jiao Liu, Qingpei Liu, Yanchun Shao, István Molnár, Fusheng Chen
B. Gillian Turgeon, Editor
Mu Li
aHubei International Scientific and Technological Cooperation Base of Traditional Fermented Foods, Key Laboratory of Environment Correlative Dietology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Mu Li
Lijing Kang
aHubei International Scientific and Technological Cooperation Base of Traditional Fermented Foods, Key Laboratory of Environment Correlative Dietology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Xiaoli Ding
aHubei International Scientific and Technological Cooperation Base of Traditional Fermented Foods, Key Laboratory of Environment Correlative Dietology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jiao Liu
aHubei International Scientific and Technological Cooperation Base of Traditional Fermented Foods, Key Laboratory of Environment Correlative Dietology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Qingpei Liu
aHubei International Scientific and Technological Cooperation Base of Traditional Fermented Foods, Key Laboratory of Environment Correlative Dietology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China
bSouthwest Center for Natural Products Research, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Yanchun Shao
aHubei International Scientific and Technological Cooperation Base of Traditional Fermented Foods, Key Laboratory of Environment Correlative Dietology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
István Molnár
bSouthwest Center for Natural Products Research, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for István Molnár
Fusheng Chen
aHubei International Scientific and Technological Cooperation Base of Traditional Fermented Foods, Key Laboratory of Environment Correlative Dietology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
B. Gillian Turgeon
Cornell University
Roles: Editor
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/mBio.02676-19
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • FIG 1
    • Open in new tab
    • Download powerpoint
    FIG 1

    A branching pathway yields MonAzPs and monasone congeners in M. ruber M7. The reactive polyketide intermediate 1 gives rise to MonAzPs on one branch (represented by rubropunctatin 7a and monascorubrin 7b) and to naphthoquinone congeners on the other branch (represented by trihydroxynaphthalene 2, monasone A [compound 3], monasone B [compound 4], tetralindione 8, and trihydroxytetralone MA-1 [compound 9]). The structures of the boxed compounds were elucidated by LC-MS/MS and NMR analysis (data for compounds 2, 3, 4, 8 and 9 are in Table S8 and Fig. S10 [19]).

  • FIG 2
    • Open in new tab
    • Download powerpoint
    FIG 2

    A conserved MrPigB regulon in MABGC-like gene clusters. (a) qRT-PCR analysis of the MABGC genes of the M. ruber M7 wild-type (WT), ΔmrpigB, and ΔmrpigC knockout strains, measured from monoculture or during cocultivation with Penicillium expansum ATCC 7861. Gene expression levels from monoculture are taken as the basis of comparison, with the means and standard deviations calculated from measurements from three biological replicates for each strain/cultivation condition shown. (b) MABGC-like gene clusters in filamentous fungal genomes. Arrows with identical colors indicate orthologous genes; white arrows show nonorthologous genes. (c) Proteins encoded by MABGC-related SM gene clusters of various fungi. Proteins highlighted in red are encoded by all MABGC-like gene clusters. Genes encoding orthologues of the MFS transporter MrPigP (highlighted in pink) are present in the majority of the MABGC-like gene clusters.

  • FIG 3
    • Open in new tab
    • Download powerpoint
    FIG 3

    Biosynthesis of monasones during in vivo and vitro enzymatic reactions with MrPigA and MrPigN. (a) Metabolite profiles (reversed-phase HPLC traces recorded at 280 nm with a photodiode array detector) of fermentation extracts of A. oryzae M-2-3 expressing the indicated MABGC genes. (b) In vitro enzymatic assays with the indicated purified MABGC enzymes (reversed-phase HPLC traces recorded at 280 nm with a photodiode array detector). (c) Quantification of trihydroxynaphthalene 2 in enzymatic reactions with the indicated enzymes after 10 min or 30 min. Yields of compound 2 are shown in micrograms per milliliter as the means ± standard deviations (SDs) from three independent experiments of three replicates each, n = 9. Statistical analysis with Student's t test revealed that there was a significant difference between group MrPigA and group MrPigA+MrPigN at P < 0.05.

  • FIG 4
    • Open in new tab
    • Download powerpoint
    FIG 4

    MrPigH-mediated reductive transformation of monasones. (a) Proposed metabolic grid for the enzymatic reduction of monasone A (compound 3) by MrPigH and MrPigC or a similar ketoreductase under aerobic or anaerobic conditions. The structures of the boxed compounds were elucidated by LC-HRMS/MS and NMR analysis (Table S8 and Fig. S10 [19]). (b) Time course analysis of the reduction of monasone A (compound 3) into compounds 10 and 11 by recombinant MrPigH under aerobic conditions. (c) Time course analysis of the reduction of monasone A (compound 3) into compounds 14 and 17 by MrPigH under anaerobic conditions. (d) Reduction of monasone B (compound 4) by recombinant MrPigH into compound 12 under aerobic conditions and to compound 16 under anaerobic conditions. Reconstituted enzymatic reactions were performed in HEPES buffer (pH 7.0) containing 1.5 mM substrate and 1.5 mM NADPH at 30°C, and metabolites were detected by reversed-phase HPLC at 280 nm with a photodiode array detector.

  • FIG 5
    • Open in new tab
    • Download powerpoint
    FIG 5

    MrPigP is an inducible naphthoquinone transporter. (a) Growth of wild-type (WT) M. ruber M7, the ΔmrpigP knockout mutant, and the mrpigC-complemented knockout strain (CΔmrpigP) on PDA plates containing 32 μg/ml or 64 μg/ml monasone A (compound 3) at 30°C for 5 days. (b) MICs of the WT, ΔmrpigP, and CΔmrpigP strains evaluated after cultivation in PDB at 30°C for 5 days. (c) Flux assay measuring the intracellular concentration of monasone A (compound 3) in the WT, ΔmrpigP, and CΔmrpigP strains after immersion in 32 μg/ml monasone A for 6 h. Statistical analysis using Student's t test revealed that there was a significant difference (P < 0.05) between group ΔmrpigP and either group WT or group CΔmrpigP. Groups WT and CΔmrpigP were not significantly different. (d) Relative transcription levels of the mrpigP gene after growth of the wild-type strain on different monasone A (compound 3) concentrations. The β-actin gene was used as an internal standard to normalize expression levels. Extracellular (e) and intracellular (f) MonAzP concentrations in the WT and ΔmrpigP strains. The strains were cultivated in PDB medium at 30°C for 10 days (mid-production phase). MonAzPs measured: Monc, monascin; Ank, ankaflavin; Rubt, rubropunctatin; Monb, monascorubrin; Rubm, rubropunctamine; Monm, monascorubramine. Statistical analysis using Student's t test revealed no significant differences between groups WT and ΔmrpigP for any of these compounds. Data from all quantitative experiments are shown as the means ± SDs from three independent experiments of three replicates each, n = 9.

  • FIG 6
    • Open in new tab
    • Download powerpoint
    FIG 6

    Model for monasone biosynthesis, export, and reductive detoxification in M. ruber M7. Thick black arrows show steps in monasone biosynthesis and recycling; thin black arrows indicate steps in MonAzP biosynthesis; blue arrows represent monasone export; dotted and solid red arrows show aerobic and anaerobic monasones detoxification steps, respectively; black block arrows indicate genes for monasone and/or MonAzP biosynthesis and resistance; gray block arrows show genes involved only in MonAzP biosynthesis.

PreviousNext
Back to top
Download PDF
Citation Tools
Monasone Naphthoquinone Biosynthesis and Resistance in Monascus Fungi
Mu Li, Lijing Kang, Xiaoli Ding, Jiao Liu, Qingpei Liu, Yanchun Shao, István Molnár, Fusheng Chen
mBio Feb 2020, 11 (1) e02676-19; DOI: 10.1128/mBio.02676-19

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this mBio article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Monasone Naphthoquinone Biosynthesis and Resistance in Monascus Fungi
(Your Name) has forwarded a page to you from mBio
(Your Name) thought you would be interested in this article in mBio.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Monasone Naphthoquinone Biosynthesis and Resistance in Monascus Fungi
Mu Li, Lijing Kang, Xiaoli Ding, Jiao Liu, Qingpei Liu, Yanchun Shao, István Molnár, Fusheng Chen
mBio Feb 2020, 11 (1) e02676-19; DOI: 10.1128/mBio.02676-19
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • INTRODUCTION
    • RESULTS
    • DISCUSSION
    • MATERIALS AND METHODS
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

Monascus spp.
naphthoquinone
nested biosynthetic pathway
supercluster
resistance mechanism

Related Articles

Cited By...

About

  • About mBio
  • Editor in Chief
  • Board of Editors
  • AAM Fellows
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Author Warranty
  • Article Types
  • Ethics
  • Contact Us

Follow #mBio

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Online ISSN: 2150-7511