摘要
L-色氨酸是芳香族氨基酸的一种,被广泛应用于医药、食品和饲料等领域。大肠杆菌磷酸烯醇式丙酮酸-糖磷酸转移酶系统(PTS系统)在葡萄糖转运和磷酸化过程中起重要作用,是糖代谢基因表达调控的核心。利用Red同源重组系统,构建包含两类典型PTS系统突变(ptsHIcrr^-glf-glk^+和ptsG^-)的L-色氨酸生产菌,并对相关菌株进行补料分批发酵研究。结果表明,不同类型PTS系统突变对菌体生长、L-色氨酸产量、糖酸转化率及副产物生成均有较大影响。与出发菌相比,ptsHIcrr^-glf-glk^+突变株最高OD_(600)达到125,提高47.0%,产酸38.5 g/L,提高25.9%,糖酸转化率16.7%,提高26.5%,乙酸生成略有增加;ptsG^-突变株最高OD_(600)达到100,提高17.6%,产酸33.4 g/L,提高9.4%,糖酸转化率15.5%,提高17.4%,乙酸生成略有减少。对葡萄糖转运系统的进一步研究将为大肠杆菌合成L-色氨酸效率的提升提供帮助。
L-tryptophan, one of the aromatic amino acids, is widely used in the fields of medicine, food and feed additives. The phosphoenolpyruvate-carbohydrate phosphotransferase system (PTS) plays an important role in glucose transport and phosphorylation in Escherichia coll. PTS-mediated regulation dominates the carbohydrates' uptake and metabolism in E. coli. We constructed L-tryptophan-producing bacteria containing two typical PTS mutations (ptsHIcrr- glf-glk+ and ptsG-) by Red homologous recombination system, and studied in 50 L jar fermenter using fed-batch fermentation. Both PTS system mutants had a great impact on the biomass (increasing 47.0% and 17.6%, respectively), L-tryptophan production (increasing 25.9% and 9.4%, respectively), glucose conversion rate (increasing 26.5% and 17.4%, respectively) and byproduct acetic acid generation (slightly increased and decreased, respectively).
出处
《生物工程学报》
CAS
CSCD
北大核心
2017年第11期1877-1882,共6页
Chinese Journal of Biotechnology
关键词
大肠杆菌
L-色氨酸
PTS系统
补料分批发酵
Escherichia coli, L-tryptophan, phosphotransferase system, fed-batch fermentation