Production of Monacolin K in Monascus pilosus: Comparison between Industrial Strains and Analysis of Its Gene Clusters

Dai, Weihua; Shao, Yanchun; Chen, Fusheng

doi:10.3390/microorganisms9040747

Cited by 8 publications

(5 citation statements)

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“…On the other hand, gene sequencing similarity of monacolin K among Monascus species was highly homologous. This finding is in agreement with the study of Dai et al, who compared the monacolin K gene cluster of seven M. pilosus and M. ruber strains (Dai et al 2021 ). The highly homologous citrinin genes in Monascus spp.…”

Section: Discussionsupporting

confidence: 93%

Identification of the high-yield monacolin K strain from Monascus spp. and its submerged fermentation using different medicinal plants

Chen

Hwang

et al. 2022

Bot Stud

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Background Medical plants confer various benefits to human health and their bioconversion through microbial fermentation can increase efficacy, reduce toxicity, conserve resources and produce new chemical components. In this study, the cholesterol-lowering monacolin K genes and content produced by Monascus species were identified. The high-yield monacolin K strain further fermented with various medicinal plants. The antioxidant and anti-inflammatory activities, red pigment and monacolin K content, total phenolic content, and metabolites in the fermented products were analyzed. Results Monacolin K was detected in Monascus pilosus (BCRC 38072), and Monascus ruber (BCRC 31533, 31523, 31534, 31535, and 33323). It responded to the highly homologous mokA and mokE genes encoding polyketide synthase and dehydrogenase. The high-yield monacolin K strain, M. ruber BCRC 31535, was used for fermentation with various medicinal plants. A positive relationship between the antioxidant capacity and total phenol content of the fermented products was observed after 60 days of fermentation, and both declined after 120 days of fermentation. By contrast, red pigment and monacolin K accumulated over time during fermentation, and the highest monacolin K content was observed in the fermentation of Glycyrrhiza uralensis, as confirmed by RT-qPCR. Moreover, Monascus-fermented medicinal plants including Paeonia lactiflora, Alpinia oxyphylla, G. uralensis, and rice were not cytotoxic. Only the product of Monascus-fermented G. uralensis significantly exhibited the anti-inflammatory capacity in a dose-dependent manner in lipopolysaccharide-induced Raw264.7 cells. The metabolites of G. uralensis with and without fermentation (60 days) were compared by LC/MS. 2,3-Dihydroxybenzoic acid, 3,4-dihydroxyphenylglycol, and 3-amino-4-hydroxybenzoate were considered to enhance the antioxidant and anti-inflammatory ability. Conclusions Given that highly homologous monacolin K and citrinin genes can be observed in Monascus spp., monacolin K produced by Monascus species without citrinin genes can be detected through the complementary methods of PCR and HPLC. In addition, the optimal fermentation time was important to the acquisition of antioxidants, red pigment and monacolin K. These bioactive substances were significantly affected by medicinal plants over fermentation time. Consequently, Monascus-fermented G. uralensis had a broad spectrum of biological activities.

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Section: Discussionsupporting

confidence: 93%

Identification of the high-yield monacolin K strain from Monascus spp. and its submerged fermentation using different medicinal plants

Chen

Hwang

et al. 2022

Bot Stud

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“…The gene sequences of the BGCs in M. ruber FWB13, M. ruber KACC 46666, M. ruber GA, and M. pilosus MS-1 were found to be highly similar to the reference BGC. M. pilosus YDJ1, M. pilosus YDJ2, and M. pilosus K104061 were isolated from commercial MK products and found to be capable of producing MK ( Dai et al, 2021 ), but antiSMASH or BLAST searches revealed incomplete BGC sequences in their genomes. MokH was absent in M. pilosus K104061, and MokE was missing from M. pilosus YDJ1’s BGC.…”

Section: Resultsmentioning

confidence: 99%

“…The phenotype-based identification schemes in Monascus were difficult to match with the results obtained by ITS, partial LSU and/or β-tubulin gene sequencing ( Patakova, 2013 ), especially in the case of single-gene locus-based phylogenetics. For example, Dai et al (2021) inferred a Neighbor-Joining tree using ITS sequences, which revealed that several isolates from M. pilosus , M. fuliginosus , M. barkeri , M. paxii , M. albidulus , M. ruber , M. purpureus , and M. fumeus were evolutionarily close in the same clade with high bootstrap values. In another clade, several strains from M. purpureus , M. rutilus , M. aurantiacus , and M. kaoliang were clustered.…”

Section: Discussionmentioning

confidence: 99%

“…More recently, DNA sequencing and molecular phylogenetics have become essential in microbiological taxonomy. ITS ( Dai et al, 2021 ), LSU ( He et al, 2020 ; Higa et al, 2020 ), β-tubulin ( Tong et al, 2022 ), calmodulin ( He et al, 2020 ; Ruiz and Radwan, 2021 ) are frequently used for accurate species identification of close relatives among Monascus spp. However, phylogenetic delineation of this genus was complicated by gene region inconsistency and low support for internal nodes.…”

Section: Introductionmentioning

confidence: 99%

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Insight into the phylogeny and metabolic divergence of Monascus species (M. pilosus, M. ruber, and M. purpureus) at the genome level

et al. 2023

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BackgroundSpecies of the genus Monascus are economically important and widely used in the production of food colorants and monacolin K. However, they have also been known to produce the mycotoxin citrinin. Currently, taxonomic knowledge of this species at the genome level is insufficient.MethodsThis study presents genomic similarity analyses through the analysis of the average nucleic acid identity of the genomic sequence and the whole genome alignment. Subsequently, the study constructed a pangenome of Monascus by reannotating all the genomes and identifying a total of 9,539 orthologous gene families. Two phylogenetic trees were constructed based on 4,589 single copy orthologous protein sequences and all the 5,565 orthologous proteins, respectively. In addition, carbohydrate active enzymes, secretome, allergic proteins, as well as secondary metabolite gene clusters were compared among the included 15 Monascus strains.ResultsThe results clearly revealed a high homology between M. pilosus and M. ruber, and their distant relationship with M. purpureus. Accordingly, all the included 15 Monascus strains should be classified into two distinctly evolutionary clades, namely the M. purpureus clade and the M. pilosus-M. ruber clade. Moreover, gene ontology enrichment showed that the M. pilosus-M. ruber clade had more orthologous genes involved with environmental adaptation than the M. purpureus clade. Compared to Aspergillus oryzae, all the Monascus species had a substantial gene loss of carbohydrate active enzymes. Potential allergenic and fungal virulence factor proteins were also found in the secretome of Monascus. Furthermore, this study identified the pigment synthesis gene clusters present in all included genomes, but with multiple nonessential genes inserted in the gene cluster of M. pilosus and M. ruber compared to M. purpureus. The citrinin gene cluster was found to be intact and highly conserved only among M. purpureus genomes. The monacolin K gene cluster was found only in the genomes of M. pilosus and M. ruber, but the sequence was more conserved in M. ruber.ConclusionThis study provides a paradigm for phylogenetic analysis of the genus Monascus, and it is believed that this report will lead to a better understanding of these food microorganisms in terms of classification, metabolic differentiation, and safety.

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“…PRJEB20852), and M. pilosus MS-1, YDJ-2, YDJ-1, and K104061 (accession no. PRJNA718072; [ 13 ]), M. purpureus YY-1 (accession no. PRJNA453427) and M. purpureus NRRL 1596 (BCRC 31542 T ; accession no.…”

Section: Methodsmentioning

confidence: 99%

Investigation of monacolin K, yellow pigments, and citrinin production capabilities of Monascus purpureus and Monascus ruber (Monascus pilosus)

Lin¹,

Chiu²,

Chen³

et al. 2023

Journal of Food and Drug Analysis

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Red mold rice (RMR) is a traditional Chinese medicine prepared using Monascus fermentation. Monascus ruber ( pilosus ) and Monascus purpureus have a long history of use as food and medicine. As an economically important starter culture, the relationship between the taxonomy of Monascus and production capabilities of secondary metabolites is crucial for the Monascus food industry. In this study, monacolin K, monascin, ankaflavin, and citrinin production by M. purpureus and M. ruber were genomically and chemically investigated. Our findings suggest that M. purpureus can produce monascin and ankaflavin in a correlated manner, whereas M. ruber produces monascin with minimum ankaflavin. M. purpureus is capable of producing citrinin; however, it is unlikely able to produce monacolin K. In contrast, M. ruber produces monacolin K, but not citrinin. We suggest that the current monacolin K content-related regulation of Monascus food should be revised, and labeling of Monascus species should be considered.

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Production of Monacolin K in Monascus pilosus: Comparison between Industrial Strains and Analysis of Its Gene Clusters

Cited by 8 publications

References 50 publications

Identification of the high-yield monacolin K strain from Monascus spp. and its submerged fermentation using different medicinal plants

Identification of the high-yield monacolin K strain from Monascus spp. and its submerged fermentation using different medicinal plants

Insight into the phylogeny and metabolic divergence of Monascus species (M. pilosus, M. ruber, and M. purpureus) at the genome level

Investigation of monacolin K, yellow pigments, and citrinin production capabilities of Monascus purpureus and Monascus ruber (Monascus pilosus)

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