两种甲醇脱氢酶在食甲基杆菌甲醇代谢中的作用  

Role of Two Methanol Dehydrogenases in Methanol Metabolism of Methylobacterium

作　　者：郭雅卿 王东澍 朱力 张惟材 刘颖[1] 王恒樑 GUO Yaqing;WANG Dongshu;ZHU Li;ZHANG Weicai;LIU Ying;WANG Hengliang(College of Food Engineering,Harbin University of Commerce,Harbin 150028,China;State Key Laboratory of Pathogen and Biosecurity,Beijing Institute of Biotechnology,Academy of Military Medical Sciences,Beijing 100071,China)

机构地区：[1]哈尔滨商业大学食品工程学院,黑龙江哈尔滨150028 [2]军事医学研究院生物工程研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《食品工业科技》2024年第10期126-132,共7页Science and Technology of Food Industry

摘　　要：本研究以食葡糖食甲基杆菌MP688(Methylobacterium glucophilum MP688)的吡咯喹啉醌(pyrroloquino-line quinone,PQQ)高产突变株J1-1为研究对象,通过生物信息学分析从基因组上找到两个甲醇脱氢酶(methanol dehydrogenase,MDH)基因mpq0771和mpq2496。考察了Ca^(2+)及La^(3+)对菌体生长、PQQ合成、MDH表达及酶活力的影响,并研究两种酶在甲醇代谢中的作用。结果显示:Ca^(2+)和La^(3+)均能促进J1-1菌体生长,且在菌体快速生长的情况下导致PQQ合成下降;Ca^(2+)存在时,菌株J1-1中mpq0771表达量较高,添加La^(3+)后mpq0771表达受到阻遏,但增强了mpq2496表达,菌体MDH总活力也有所提高;敲除菌J1-1Δmpq0771不能利用甲醇生长,添加La^(3+)后可利用甲醇生长并能合成PQQ,且能检测到MDH活性。mpq0771编码产物是Ca^(2+)依赖的MDH,mpq2496编码产物是严格依赖La^(3+)的MDH,且La^(3+)能够诱导mpq2496表达并替代mpq0771发挥甲醇脱氢酶的作用。In this study,the pyrroloquinoline quinone(PQQ)high-yield mutant J1-1 of Methylobacterium glucophilum MP688 was used to do bioinformatics analysis to extract the two methanol dehydrogenase(MDH)genes from the genome,mpq0771 and mpq2496.The effects of Ca^(2+)and La^(3+)on bacterial growth,PQQ synthesis,MDH expression,and enzyme activity were explored,as well as the roles of the two enzymes in methanol metabolism.The results showed that Ca^(2+)and La^(3+)both increased J1-1 bacteriophage growth and decreased PQQ synthesis in the context of fast viral development.In the presence of Ca^(2+),mpq0771 expression increased in strain J1-1.La^(3+)inhibited mpq0771 expression while promoting mpq2496 expression and increasing total bacterial MDH viability.The knockout bacteria J1-1Δmpq0771 could not grow in methanol,but with the addition of La^(3+),it could grow in methanol and synthesize PQQ,and MDH activity was identified.The product encoded by mpq0771 was a Ca^(2+)-dependent MDH,and the product encoded by mpq2496 was a La^(3+)-dependent MDH,and La^(3+)could increase the production of mpq2496 and replace mpq0771 as a methanol dehydrogenase.

关 键 词：食甲基杆菌 甲醇脱氢酶 吡咯喹啉醌 La^(3+) 

分 类 号：TS201.3[轻工技术与工程—食品科学] Q939.9[轻工技术与工程—食品科学与工程]