Research Institute of Innovative Technology for the Earth, 9-2 Kizugawadai, Kizugawa, Kyoto, Japan.
Appl Microbiol Biotechnol. 2010 Apr;86(4):1057-66. doi: 10.1007/s00253-009-2372-2. Epub 2009 Dec 10.
Wild-type Corynebacterium glutamicum produced 0.6 g l(-1) xylitol from xylose at a productivity of 0.01 g l(-1) h(-1) under oxygen deprivation. To increase this productivity, the pentose transporter gene (araE) from C. glutamicum ATCC31831 was integrated into the C. glutamicum R chromosome. Consequent disruption of its lactate dehydrogenase gene (ldhA), and expression of single-site mutant xylose reductase from Candida tenuis (CtXR (K274R)) resulted in recombinant C. glutamicum strain CtXR4 that produced 26.5 g l(-1) xylitol at 3.1 g l(-1) h(-1). To eliminate possible formation of toxic intracellular xylitol phosphate, genes encoding xylulokinase (XylB) and phosphoenolpyruvate-dependent fructose phosphotransferase (PTS(fru)) were disrupted to yield strain CtXR7. The productivity of strain CtXR7 increased 1.6-fold over that of strain CtXR4. A fed-batch 21-h CtXR7 culture in mineral salts medium under oxygen deprivation yielded 166 g l(-1) xylitol at 7.9 g l(-1) h(-1), representing the highest bacterial xylitol productivity reported to date.
野生型谷氨酸棒杆菌在缺氧条件下,以 0.01 g l(-1) h(-1)的产率从木糖生产 0.6 g l(-1)木糖醇。为了提高这一生产力,将谷氨酸棒杆菌 ATCC31831 的戊糖转运蛋白基因(araE)整合到谷氨酸棒杆菌 R 染色体上。随后敲除其乳酸脱氢酶基因(ldhA),并表达来自脆弱拟杆菌的单点突变木糖还原酶(CtXR(K274R)),导致重组谷氨酸棒杆菌菌株 CtXR4 以 3.1 g l(-1) h(-1)的产率生产 26.5 g l(-1)木糖醇。为了消除可能形成的有毒细胞内木糖醇磷酸盐,敲除编码木酮糖激酶(XylB)和磷酸烯醇丙酮酸依赖性果糖磷酸转移酶(PTS(fru))的基因,得到菌株 CtXR7。与菌株 CtXR4 相比,菌株 CtXR7 的生产力提高了 1.6 倍。在缺氧条件下,矿物盐培养基中进行 21 小时的 CtXR7 分批补料培养,以 7.9 g l(-1) h(-1)的产率生产 166 g l(-1)木糖醇,这是迄今为止报道的最高细菌木糖醇生产力。