Inactivation of MrPigH in Monascus ruber M7 Results in its Monascus Pigments Increase and Citrinin Decrease with mrpyrG Selection Marker

Monascus pigments (MPs) have been used as food colorants for several centuries in Asian countries and nowadays in the whole world via Asian catering. The MPs biosynthetic pathway has been well-illustrated, however, the functions of a few genes including mrpigH in the MPs gene cluster of M. ruber M7 are still unclear. In current study, mrpigH was disrupted in Δmrlig4ΔmrpyrG, a highly efficient gene modification system, using mrpyrG as a selection marker, and ΔmrpigHΔmrlig4ΔmrpyrG::mrpyrG and ΔmrpigHΔmrlig4ΔmrpyrG have been obtained. Subsequently, their morphologies, biomasses, MPs and citrinin (CIT) production were analyzed, respectively. These results have revealed that the deletion of mrpigH has significant effects on the morphology and growth of M. ruber M7. Moreover, compared with M. ruber M7, the yields of MPs and CIT were drastically increased and decreased in mrpigH mutants, respectively.

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Inactivation of mrpigH Gene in Monascus ruber M7 Results in Increased Monascus Pigments and Decreased Citrinin with mrpyrG Selection Marker

Journal of Fungi ◽

10.3390/jof7121094 ◽

2021 ◽

Vol 7 (12) ◽

pp. 1094

Author(s):

Li Li ◽

Na Xu ◽

Fusheng Chen

Keyword(s):

Gene Cluster ◽

Biosynthetic Pathway ◽

Selection Marker ◽

Asian Countries ◽

Gene Modification ◽

Monascus Ruber ◽

Modification System ◽

Monascus Pigments ◽

Efficient Gene ◽

The World

Monascus pigments (MPs) have been used as food colorants for several centuries in Asian countries and are currently used around the world via Asian catering. The MPs biosynthetic pathway has been well-illustrated; however, the functions of a few genes including mrpigH in the MPs gene cluster of M. ruber M7 are still unclear. In the current study, mrpigH was disrupted in Δmrlig4ΔmrpyrG, a highly efficient gene modification system, using mrpyrG as a selection marker, and ΔmrpigHΔmrlig4ΔmrpyrG::mrpyrG and ΔmrpigHΔmrlig4ΔmrpyrG have been obtained. Subsequently, their morphologies, biomasses, MPs and citrinin (CIT) production were analyzed, respectively. These results have revealed that the deletion of mrpigH has significant effects on the morphology and growth of M. ruber M7. Moreover, compared with M. ruber M7, the yields of MPs and CIT were drastically increased and decreased in mrpigH mutants, respectively.

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Effects of mrpigG on Development and Secondary Metabolism of Monascus ruber M7

Journal of Fungi ◽

10.3390/jof6030156 ◽

2020 ◽

Vol 6 (3) ◽

pp. 156

Author(s):

Li Li ◽

Fusheng Chen

Keyword(s):

Biosynthetic Pathway ◽

Gene Deletion ◽

The Other ◽

Side Chain ◽

Asian Countries ◽

Monascus Ruber ◽

Monascus Pigments ◽

The World ◽

Carbon Side Chain ◽

Yellow Pigments

Monascus pigments (MPs) have been used as food colorants for several centuries in Asian countries and are now used throughout the world via Asian catering. The MP biosynthetic pathway has been well-illustrated, but the functions of a few genes, including mrpigG, in the MP gene cluster are still unclear. In the current study, in order to investigate the function of mrpigG in M. ruber M7, gene deletion (ΔmrpigG), complementation (ΔmrpigG::mrpigG) and overexpression (M7::PtrpC-mrpigG) mutants were successfully obtained. The morphologies and biomasses, as well as the MP and citrinin production, of these mutants were analyzed. The results revealed that the disruption, complementation and overexpression of mrpigG showed no apparent defects in morphology, biomass or citrinin production (except MP production) in ΔmrpigG compared with M. ruber M7. Although the MP profiles of ΔmrpigG and M. ruber M7 were almost the same—with both having four yellow pigments, two orange pigments (OPs) and two red pigments (RPs)—their yields were decreased in ΔmrpigG to a certain extent. Particularly, the content of rubropunctatin (an OP) and its derivative rubropunctamine (an RP) in ΔmrpigG, both of which have a five-carbon side chain, accounted for 57.7%, and 22.3% of those in M. ruber M7. On the other hand, monascorubrin (an OP) and its derivative monascorubramine (an RP), both of which have a seven-carbon side chain, were increased by 1.15 and 2.55 times, respectively, in ΔmrpigG compared with M. ruber M7. These results suggest that the MrPigG protein may preferentially catalyze the biosynthesis of MPs with a five-carbon side chain.

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Effect of Static Magnetic Field on Monascus ruber M7 Based on Transcriptome Analysis

Journal of Fungi ◽

10.3390/jof7040256 ◽

2021 ◽

Vol 7 (4) ◽

pp. 256

Author(s):

Shuyan Yang ◽

Hongyi Zhou ◽

Weihua Dai ◽

Juan Xiong ◽

Fusheng Chen

Keyword(s):

Magnetic Field ◽

Static Magnetic Field ◽

Morphological Characteristics ◽

Mapk Signaling ◽

Mitogen Activated Protein Kinase ◽

Primary Metabolism ◽

Monascus Ruber ◽

Growing Period ◽

Monascus Pigments ◽

Mitogen Activated Protein

The effects of a static magnetic field (SMF) on Monascus ruber M7 (M. ruber M7) cultured on potato dextrose agar (PDA) plates under SMF treatment at different intensities (5, 10, and 30 mT) were investigated in this paper. The results revealed that, compared with the control (CK, no SMF treatment), the SMF at all tested intensities did not significantly influence the morphological characteristics of M. ruber M7, while the intracellular and extracellular Monascus pigments (MPs) and extracellular citrinin (CIT) of M. ruber M7 were increased at 10 and 30 mT SMF but there was no impact on the MPs and CIT at 5 mT SMF. The transcriptome data of M. ruber M7 cultured at 30 mT SMF on PDA for 3 and 7 d showed that the SMF could increase the transcriptional levels of some relative genes with the primary metabolism, including the carbohydrate metabolism, amino acid metabolism, and lipid metabolism, especially in the early growing period (3 d). SMF could also affect the transcriptional levels of the related genes to the biosynthetic pathways of MPs, CIT, and ergosterol, and improve the transcription of the relative genes in the mitogen-activated protein kinase (MAPK) signaling pathway of M. ruber M7. These findings provide insights into a comprehensive understanding of the effects of SMF on filamentous fungi.

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A Gene Cluster Containing Two Fungal Polyketide Synthases Encodes the Biosynthetic Pathway for a Polyketide, Asperfuranone, inAspergillus nidulans

Journal of the American Chemical Society ◽

10.1021/ja8088185 ◽

2009 ◽

Vol 131 (8) ◽

pp. 2965-2970 ◽

Cited By ~ 199

Author(s):

Yi-Ming Chiang ◽

Edyta Szewczyk ◽

Ashley D. Davidson ◽

Nancy Keller ◽

Berl R. Oakley ◽

...

Keyword(s):

Gene Cluster ◽

Biosynthetic Pathway ◽

Polyketide Synthases

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The fungal gene cluster for biosynthesis of the antibacterial agent viriditoxin

10.1101/601716 ◽

2019 ◽

Cited By ~ 1

Author(s):

Andrew S. Urquhart ◽

Jinyu Hu ◽

Yit-Heng Chooi ◽

Alexander Idnurm

Keyword(s):

Secondary Metabolites ◽

Gene Cluster ◽

Gene Disruption ◽

Biosynthetic Pathway ◽

Fluorescent Protein ◽

Model Species ◽

Paecilomyces Variotii ◽

Bioinformatic Approaches ◽

Green Fluorescent ◽

Fungal Gene

AbstractBackgroundViriditoxin is one of the ‘classical’ secondary metabolites produced by fungi and that has antibacterial and other activities; however, the mechanism of its biosynthesis has remained unknown.ResultsHere, a gene cluster responsible for its synthesis was identified, using bioinformatic approaches from two species that produce viriditoxin and then through gene disruption and metabolite profiling. All eight genes in the cluster inPaecilomyces variotiiwere mutated, revealing their roles in the synthesis of this molecule and establishing its biosynthetic pathway which includes an interesting Baeyer-Villiger monooxygenase catalyzed reaction. Additionally, a candidate catalytically-inactive hydrolase was identified as being required for the stereoselective biosynthesis of (M)-viriditoxin. The localization of two proteins were assessed by fusing these proteins to green fluorescent protein, revealing that at least two intracellular structures are involved in the compartmentalization of the synthesis steps of this metabolite.ConclusionsThe full pathway for synthesis of viriditoxin was established by a combination of genomics, bioinformatics, gene disruption and chemical analysis processes. Hence, this work reveals the basis for the synthesis of an understudied class of fungal secondary metabolites and provides a new model species for understanding the synthesis of biaryl compounds with a chiral axis.

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Biosynthesis of the uridine-derived nucleoside antibiotic A-94964: identification and characterization of the biosynthetic gene cluster provide insight into the biosynthetic pathway

Organic & Biomolecular Chemistry ◽

10.1039/c8ob02765j ◽

2019 ◽

Vol 17 (3) ◽

pp. 461-466 ◽

Cited By ~ 5

Author(s):

Taro Shiraishi ◽

Makoto Nishiyama ◽

Tomohisa Kuzuyama

Keyword(s):

Gene Cluster ◽

In Silico ◽

Biosynthetic Pathway ◽

Gene Deletion ◽

Biosynthetic Gene Cluster ◽

Biosynthetic Gene ◽

Identification And Characterization ◽

Insight Into

The biosynthetic pathway of the uridine-derived nucleoside antibiotic A-94964 was proposed via in silico analysis coupled with gene deletion experiments.

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Biosynthesis of Fusarubins Accounts for Pigmentation of Fusarium fujikuroi Perithecia

Applied and Environmental Microbiology ◽

10.1128/aem.00823-12 ◽

2012 ◽

Vol 78 (12) ◽

pp. 4468-4480 ◽

Cited By ~ 109

Author(s):

Lena Studt ◽

Philipp Wiemann ◽

Karin Kleigrewe ◽

Hans-Ulrich Humpf ◽

Bettina Tudzynski

Keyword(s):

Secondary Metabolites ◽

Gene Cluster ◽

Biosynthetic Pathway ◽

Polyketide Synthase ◽

Biological Function ◽

Fusarium Fujikuroi ◽

Diverse Group ◽

Fruiting Bodies ◽

Content Type ◽

Encoding Gene

ABSTRACTFusarium fujikuroiproduces a variety of secondary metabolites, of which polyketides form the most diverse group. Among these are the highly pigmented naphthoquinones, which have been shown to possess different functional properties for the fungus. A group of naphthoquinones, polyketides related to fusarubin, were identified inFusariumspp. more than 60 years ago, but neither the genes responsible for their formation nor their biological function has been discovered to date. In addition, although it is known that the sexual fruiting bodies in which the progeny of the fungus develops are darkly colored by a polyketide synthase (PKS)-derived pigment, the structure of this pigment has never been elucidated. Here we present data that link the fusarubin-type polyketides to a defined gene cluster, which we designatefsr, and demonstrate that the fusarubins are the pigments responsible for the coloration of the perithecia. We studied their regulation and the function of the single genes within the cluster by a combination of gene replacements and overexpression of the PKS-encoding gene, and we present a model for the biosynthetic pathway of the fusarubins based on these data.

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A GENETIC AND BIOCHEMICAL STUDY OF HISTIDINE BIOSYNTHESIS IN MICROCOCCUS LUTEUS

Genetics ◽

10.1093/genetics/79.3.361 ◽

1975 ◽

Vol 79 (3) ◽

pp. 361-376

Author(s):

Caroline Kane-Falce ◽

Wesley E Kloos

Keyword(s):

Gene Cluster ◽

Biosynthetic Pathway ◽

Gene Sequence ◽

Biochemical Study ◽

Micrococcus Luteus ◽

Gene Clusters ◽

Growth Responses ◽

Histidine Biosynthesis ◽

Marshall Test ◽

Linkage Relationships

ABSTRACT Histidine auxotrophs of Micrococcus luteus strain ATCC 27141 were induced by treatment of the parent strain with N-methyl-N′-nitro-N-nitro-soguanidine. Auxotrophs were biochemically characterized by examining culture accumulations of histidine intermediates, using paper chromatography and the Bratton-Marshall test, and growth responses to L-histidinol. his(IG) mutants failed to accumulate Pauly-positive imidazoles; his(EAHF) mutants accumulated 5-amino-1-ribosyl-4-imidazole carboxamide; hisB mutants accumulated imidazoleglycerol; hisC mutants accumulated imidazoleacetol; hisD mutants accumulated histidinol. L-histidinol failed to stimulate the growth of hisD mutants, but did stimulate all other histidine mutants, blocked at earlier steps in the biosynthetic pathway. In addition, imidazoleglycerol phosphate dehydrase activity was assayed in representative mutants of each class. hisB mutants lacked activity for this enzyme.—Two-point, three-point, and cotransformation analyses resolved linkage relationships of histidine genes and in two gene clusters aided in determining their sequences. Histidine biosynthetic genes exist in at least four separate, unlinked regions of the chromosome. One histidine gene cluster is closely linked to a tryptophan gene cluster and appears to be contiguous in the sequence his(IG)-his(EAHF)-trpE-trpC-trpB-trpA. A second and unlinked histidine cluster has the tentative gene sequence his(EAHF)-hisB-hisC-his(EAHF). The hisD gene and an unclassified mutant site his-94 are not linked to any of the other histidine genes examined in this study or to each other.

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