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
郭雅卿 1王东澍 2朱力 2张惟材 2刘颖 1王恒樑2
作者信息
- 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
- 折叠
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Ca2+及La3+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:Ca2+和La3+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;Ca2+存在时,菌株J1-1中mpq0771表达量较高,添加La3+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La3+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 Ca2+依赖的 MDH,mpq2496编码产物是严格依赖La3+的MDH,且La3+能够诱导mpq2496表达并替代mpq0771发挥甲醇脱氢酶的作用.
Abstract
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 Ca2+ and La3+ 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 Ca2+ and La3+ both increased J1-1 bacteriophage growth and decreased PQQ synthesis in the context of fast viral development.In the presence of Ca2+,mpq0771 expression increased in strain J1-1.La3+ 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 La3+,it could grow in methanol and synthesize PQQ,and MDH activity was identified.The product encoded by mpq0771 was a Ca2+-dependent MDH,and the product encoded by mpq2496 was a La3+-dependent MDH,and La3+ 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/La3+Key words
Methylobacterium/methanol dehydrogenase/pyrroloquinoline quinone/lanthanide引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据