CAJ | 학술논문

构建了共表达烟酸转磷酸核糖激酶（ NAPRTase）和丙酮酸羧化酶（ PYC）的重组质粒pTrc99a pncB pyc，并考察了重组菌 E． coli NZN111/pTrc99a pncB pyc 生产丁二酸的能力。结果表明：重组菌 NZN111/pTrc99a pncB pyc的NAPRTase和PYC的比酶活达到最高，分别为20?75和1?04 U/mg，同时，辅酶NADH、NAD＋及NAD （ H）总量达到最高。厌氧摇瓶发酵结果：48 h能够消耗17?5 g/L的葡萄糖生成14?08 g/L的丁二酸，而丙酮酸的产量大幅度降低，仅为0?11 g/L。本研究为基因工程菌大肠杆菌厌氧条件下发酵生产丁二酸提供了一定的基础。
구건료공표체연산전린산핵당격매（ NAPRTase）화병동산최화매（ PYC）적중조질립pTrc99a pncB pyc，병고찰료중조균 E． coli NZN111/pTrc99a pncB pyc 생산정이산적능력。결과표명：중조균 NZN111/pTrc99a pncB pyc적NAPRTase화PYC적비매활체도최고，분별위20?75화1?04 U/mg，동시，보매NADH、NAD＋급NAD （ H）총량체도최고。염양요병발효결과：48 h능구소모17?5 g/L적포도당생성14?08 g/L적정이산，이병동산적산량대폭도강저，부위0?11 g/L。본연구위기인공정균대장간균염양조건하발효생산정이산제공료일정적기출。
The recombinant plasmid pTrc99a-pncB-pyc, co-expressing nicotinic acid phosphoribosy ltransferase ( NAPRTase ) and pyruvate carboxylase ( PYC ) , was constructed and the capability of succinic acid production in E. coli NZN111/pTrc99a-pncB-pyc was investigated. The results showed that the specific enzyme activities of NAPRTase and PYC in E. coli NZN111/pTrc99a-pncB-pyc reached the highest, which were 20?75 and 1?04 U/mg, respectively. At the same time, the total amount of NADH, NAD+, and NAD ( H ) reached the highest. The fermentation results in the sealed bottles showed that 17?5 g/L glucose was consumed and 14?08 g/L was produced at 48 h and the accumulation of pyruvic acid was decreased to 0?11 g/L. The study provided a basis for the production of succinic acid in the engineered E.coli under anaerobic conditions.