{"title":"莫克 I 基因过表达促进莫纳可林 K 生产的分子机制","authors":"Zhiwei Huang, Lishi Xiao, Wenlan Mo, Yaru Zhang, Yiyang Cai, Simei Huang, Zhiting Chen, Chuannan Long","doi":"10.3390/jof10100721","DOIUrl":null,"url":null,"abstract":"<p><p><i>Monascus</i> species are capable of producing various active metabolites, including monacolin K (MK) and pigments. Studies have shown that the overexpression of the <i>mok I</i> gene from the MK synthesis gene cluster in <i>Monascus</i> species can significantly increase MK production; however, the molecular mechanism has not yet been fully elucidated. Therefore, this study focused on the <i>mok I</i> gene of <i>Monascus pilosus</i> to construct overexpression strains of the <i>mok I</i> gene, resulting in high-yield MK production. Sixteen positive transformants were obtained, seven of which produced 9.63% to 41.39% more MK than the original strain, with no citrinin detected in any of the transformants. The qRT-PCR results revealed that the expression levels of <i>mok I</i> in the transformed strains TI-13, TI-24, and TI-25 increased by more than 50% compared to the original strain at various fermentation times, with the highest increase being 10.9-fold. Furthermore, multi-omics techniques were used to analyze the molecular mechanisms underlying enhanced MK production in transformed strains. The results indicated that <i>mok I</i> overexpression may enhance MK synthesis in <i>M. pilosus</i> by regulating the expression of key genes (such as <i>MAO</i>, <i>HPD</i>, <i>ACX</i>, and <i>PLC</i>) and the synthesis levels of key metabolites (such as delta-tocopherol and alpha-linolenic acid) in pathways linked to the biosynthesis of cofactors, the biosynthesis of unsaturated fatty acids, tyrosine metabolism, ubiquinone and other terpenoid-quinone biosynthesis, alpha-linolenic acid metabolism, and glycerophospholipid metabolism. These findings provide a theoretical basis for further study of the metabolic regulation of MK in <i>Monascus</i> species and for effectively enhancing their MK production.</p>","PeriodicalId":15878,"journal":{"name":"Journal of Fungi","volume":"10 10","pages":""},"PeriodicalIF":4.2000,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11508744/pdf/","citationCount":"0","resultStr":"{\"title\":\"Molecular Mechanism of <i>Mok I</i> Gene Overexpression in Enhancing Monacolin K Production in <i>Monascus pilosus</i>.\",\"authors\":\"Zhiwei Huang, Lishi Xiao, Wenlan Mo, Yaru Zhang, Yiyang Cai, Simei Huang, Zhiting Chen, Chuannan Long\",\"doi\":\"10.3390/jof10100721\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><i>Monascus</i> species are capable of producing various active metabolites, including monacolin K (MK) and pigments. Studies have shown that the overexpression of the <i>mok I</i> gene from the MK synthesis gene cluster in <i>Monascus</i> species can significantly increase MK production; however, the molecular mechanism has not yet been fully elucidated. Therefore, this study focused on the <i>mok I</i> gene of <i>Monascus pilosus</i> to construct overexpression strains of the <i>mok I</i> gene, resulting in high-yield MK production. Sixteen positive transformants were obtained, seven of which produced 9.63% to 41.39% more MK than the original strain, with no citrinin detected in any of the transformants. The qRT-PCR results revealed that the expression levels of <i>mok I</i> in the transformed strains TI-13, TI-24, and TI-25 increased by more than 50% compared to the original strain at various fermentation times, with the highest increase being 10.9-fold. Furthermore, multi-omics techniques were used to analyze the molecular mechanisms underlying enhanced MK production in transformed strains. The results indicated that <i>mok I</i> overexpression may enhance MK synthesis in <i>M. pilosus</i> by regulating the expression of key genes (such as <i>MAO</i>, <i>HPD</i>, <i>ACX</i>, and <i>PLC</i>) and the synthesis levels of key metabolites (such as delta-tocopherol and alpha-linolenic acid) in pathways linked to the biosynthesis of cofactors, the biosynthesis of unsaturated fatty acids, tyrosine metabolism, ubiquinone and other terpenoid-quinone biosynthesis, alpha-linolenic acid metabolism, and glycerophospholipid metabolism. These findings provide a theoretical basis for further study of the metabolic regulation of MK in <i>Monascus</i> species and for effectively enhancing their MK production.</p>\",\"PeriodicalId\":15878,\"journal\":{\"name\":\"Journal of Fungi\",\"volume\":\"10 10\",\"pages\":\"\"},\"PeriodicalIF\":4.2000,\"publicationDate\":\"2024-10-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11508744/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Fungi\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.3390/jof10100721\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Fungi","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.3390/jof10100721","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
莫纳科菌类能够产生各种活性代谢产物,包括莫纳可林 K(MK)和色素。研究表明,过表达莫纳可林合成基因簇中的 mok I 基因可显著提高莫纳可林的产量,但其分子机制尚未完全阐明。因此,本研究聚焦于Monascus pilosus的mok I基因,构建mok I基因的过表达菌株,从而获得高产的MK产量。研究获得了 16 个阳性转化株,其中 7 个转化株的 MK 产量比原始菌株高出 9.63% 至 41.39%,所有转化株均未检测到柠檬蛋白。qRT-PCR 结果显示,在不同的发酵时间,转化菌株 TI-13、TI-24 和 TI-25 中 mok I 的表达水平比原菌株提高了 50%以上,最高提高了 10.9 倍。此外,研究人员还利用多组学技术分析了转化菌株MK产量增加的分子机制。结果表明,mok I 的过表达可通过调控 M. pilosus 菌株的表达来提高 MK 的合成。结果表明,mok I 的过表达可通过调节关键基因(如 MAO、HPD、ACX 和 PLC)的表达和关键代谢产物(如 delta-生育酚和α-亚麻酸)在辅助因子生物合成、不饱和脂肪酸生物合成、酪氨酸代谢、泛醌和其他萜类-醌类生物合成、α-亚麻酸代谢和甘油磷脂代谢等相关途径中的合成水平来提高 MK 的合成。这些发现为进一步研究莫纳树属物种的 MK 代谢调节和有效提高其 MK 产量提供了理论依据。
Molecular Mechanism of Mok I Gene Overexpression in Enhancing Monacolin K Production in Monascus pilosus.
Monascus species are capable of producing various active metabolites, including monacolin K (MK) and pigments. Studies have shown that the overexpression of the mok I gene from the MK synthesis gene cluster in Monascus species can significantly increase MK production; however, the molecular mechanism has not yet been fully elucidated. Therefore, this study focused on the mok I gene of Monascus pilosus to construct overexpression strains of the mok I gene, resulting in high-yield MK production. Sixteen positive transformants were obtained, seven of which produced 9.63% to 41.39% more MK than the original strain, with no citrinin detected in any of the transformants. The qRT-PCR results revealed that the expression levels of mok I in the transformed strains TI-13, TI-24, and TI-25 increased by more than 50% compared to the original strain at various fermentation times, with the highest increase being 10.9-fold. Furthermore, multi-omics techniques were used to analyze the molecular mechanisms underlying enhanced MK production in transformed strains. The results indicated that mok I overexpression may enhance MK synthesis in M. pilosus by regulating the expression of key genes (such as MAO, HPD, ACX, and PLC) and the synthesis levels of key metabolites (such as delta-tocopherol and alpha-linolenic acid) in pathways linked to the biosynthesis of cofactors, the biosynthesis of unsaturated fatty acids, tyrosine metabolism, ubiquinone and other terpenoid-quinone biosynthesis, alpha-linolenic acid metabolism, and glycerophospholipid metabolism. These findings provide a theoretical basis for further study of the metabolic regulation of MK in Monascus species and for effectively enhancing their MK production.
期刊介绍:
Journal of Fungi (ISSN 2309-608X) is an international, peer-reviewed scientific open access journal that provides an advanced forum for studies related to pathogenic fungi, fungal biology, and all other aspects of fungal research. The journal publishes reviews, regular research papers, and communications in quarterly issues. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. Therefore, there is no restriction on paper length. Full experimental details must be provided so that the results can be reproduced.