肺炎克雷伯氏菌3-羟基丙酸合成关键酶基因过表达与NAD^+再生的耦合  

Coupling NAD^+ regeneration with overexpression of key enzymes for 3-hydroxypropionic acid biosynthesis in Klebsiella pneumoniae

作　　者：曹宁[1] 李莎[1] 葛喜珍[2] 田平芳[1] 

机构地区：[1]北京化工大学生命科学与技术学院,北京100029 [2]北京联合大学生物化学工程学院,北京100023

出　　处：《北京化工大学学报（自然科学版）》2014年第1期74-77,共4页Journal of Beijing University of Chemical Technology(Natural Science Edition)

基　　金：国家"973"计划(2012CB725200);国家自然科学基金(20876009/21076013/21276014)

摘　　要：在肺炎克雷伯氏菌(Klebsiella pneumoniae)中超表达内源甘油脱水酶基因dhaB和醛脱氢酶基因puuC,同时引入酿酒酵母(Saccharomyces cerevisiae)的3-磷酸甘油脱氢酶基因GPD1,得到耦合3-羟基丙酸(3-HP)合成与NAD+再生的重组菌K.p(pET-pk-dhaB-puuC-GPD1)。微氧摇瓶发酵结果显示在15h时3-HP产量达到1.45g/L,是携带空载体菌的2.39倍。分析是遗传扰动影响了底物甘油的代谢流量分配,从而提高了3-羟基丙酸的产量。The glycerol dehydratase coding gene dhaB and the aldehyde dehydrogenase coding gene puuC from Klebsiella pneumoniae,and the 3-phosphoric acid glycerol dehydrogenase coding gene GPD1 from Saccharomyces cerevisiae,have been cloned by the polymerase chain reaction(PCR). Subsequently the recombinant K. pneumoniae K. p(pET-pk-dhaB-puuC-GPD1) was acquired. Under micro-oxygen fermentation conditions,it showed peak production of 3-hydroxypropionic acid(1. 45 g/ L) at 15 h,which is 2. 39 times that obtained with the strain harboring the blank plasmid pET-pk. These results suggest that overexpressing dhaB,puuC and GPD1 facilitates 3-HP biosynthesis via reallocation of the metabolic flux in K. pneumoniae.

关 键 词：肺炎克雷伯氏菌 3-羟基丙酸 甘油脱水酶 醛脱氢酶 NAD+ 

分 类 号：Q78[生物学—分子生物学]