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常压室温等离子体诱变的芽孢乳杆菌Y2-8突变体提高d-乳酸产量

Enhanced production of d-lactic acid by Sporolactobacillus sp.Y2-8 mutant generated by atmospheric and room temperature plasma.

作者信息

Sun Jiaduo, Wang Yan, Wu Bin, Bai Zhongzhong, He Bingfang

机构信息

College of Biotechnology and Pharmaceutical Engineering, Nanjing Technology University, Nanjing, Jiangsu, People's Republic of China.

出版信息

Biotechnol Appl Biochem. 2015 Mar-Apr;62(2):287-92. doi: 10.1002/bab.1267. Epub 2014 Sep 23.

Abstract

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.

摘要

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。

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