Sun Jiaduo, Wang Yan, Wu Bin, Bai Zhongzhong, He Bingfang
College of Biotechnology and Pharmaceutical Engineering, Nanjing Technology University, Nanjing, Jiangsu, People's Republic of China.
Biotechnol Appl Biochem. 2015 Mar-Apr;62(2):287-92. doi: 10.1002/bab.1267. Epub 2014 Sep 23.
To improve the production of d-lactic acid, atmospheric and room temperature plasma (ARTP) was used to generate mutations in Sporolactobacillus sp. Y2-8. An efficient mutant YBS1-5 was rapidly isolated by implanting ARTP twice with a 100 W radio-frequency power input, 10 standard liters per minute of the helium flow, and a 2 mm treatment distance. Significant improvement of d-lactic acid productivity (1.39 g L(-1) H(-1) ) by YBS1-5 was achieved, and it was 41.84% higher than the productivity (0.98 g L(-1) H(-1) ) of Y2-8. Moreover, the dry cell weight of YBS1-5 was 16.7% higher than that of Y2-8. Metabolic activities of concerned substrates related with key enzymes of d-lactic acid fermentation were analyzed by Biolog approach. Results showed that the activities of the key enzymes glucokinase and d-lactate dehydrogenase in mutant YBS1-5 were increased by approximately 45% and 66%, respectively, in comparison with those of the strain Y2-8. Fed-batch fermentation further improved the productivity; 127 g L(-1) d-lactic acid in 74 H by YBS1-5 with higher productivity (1.72 g L(-1) H(-1) ) was achieved. The subculture experiments indicated that YBS1-5 was genetically stable after eight generations.
为提高d-乳酸的产量,采用常压室温等离子体(ARTP)对芽孢乳杆菌属Y2-8进行诱变。通过以100 W射频功率输入、每分钟10标准升的氦气流速和2 mm处理距离进行两次ARTP注入,快速筛选出一株高效突变体YBS1-5。YBS1-5的d-乳酸产量显著提高(1.39 g L⁻¹ h⁻¹),比Y2-8的产量(0.98 g L⁻¹ h⁻¹)高41.84%。此外,YBS1-5的干细胞重量比Y2-8高16.7%。采用Biolog方法分析了与d-乳酸发酵关键酶相关的底物的代谢活性。结果表明,与Y2-8菌株相比,突变体YBS1-5中关键酶葡萄糖激酶和d-乳酸脱氢酶的活性分别提高了约45%和66%。补料分批发酵进一步提高了产量;YBS1-5在74小时内实现了127 g L⁻¹的d-乳酸产量,且生产率更高(1.72 g L⁻¹ h⁻¹)。传代实验表明,YBS1-5在八代后遗传稳定。