Zhen Jie, Tan Ming, Fu Xiaoping, Shu Wenju, Zhao Xingya, Yang Shibin, Xu Jianyong, Ma Yanhe, Zheng Hongchen, Song Hui
1Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, No. 32 West 7th Avenue, Tianjin Airport Economic Park, Tianjin, 300308 People's Republic of China.
2Tianjin Key Laboratory for Industrial Biological Systems and Bioprocessing Engineering, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308 People's Republic of China.
3 Biotech. 2020 Feb;10(2):49. doi: 10.1007/s13205-019-2022-z. Epub 2020 Jan 14.
A high heterologous expression of an alkaline pectate lyase (APL) pelNK93I in was obtained through optimizing the lactose feeding and fed-batch fermentation. The highest soluble APL activity produced by BL21 (pET22b-) was 10,181 U/mL which is the highest level so far. On this basis, to improve the extracellular yield of APL, optimized glycine feeding was used to achieve elevated extracellular production of pelNK93I. The highest extracellular APL activity produced by BL21 (pET22b-) was 6357 U/mL which was also relatively higher than that in previous reports. The final productivity of APL was 282.8 U/mL/h in the fermentation of BL21 (pET22b-3I) in a 10 L fermenter. Thus the current study has provided a cost-effective method for the over-expression and preparation of alkaline pectate lyase pelNK93I for its industrial applications. Moreover, pelNK93I (4 U/mL) used for bioscouring increased cottonseed husk removal and radial capillary effect of cotton fabric by 37.63% and 47.06%, respectively, making it a promising enzyme in green textile technology.
通过优化乳糖补料和分批补料发酵,实现了碱性果胶酸裂解酶(APL)pelNK93I在大肠杆菌中的高异源表达。大肠杆菌BL21(pET22b-)产生的最高可溶性APL活性为10181 U/mL,这是迄今为止的最高水平。在此基础上,为提高APL的胞外产量,采用优化的甘氨酸补料实现了pelNK93I胞外产量的提高。大肠杆菌BL21(pET22b-)产生的最高胞外APL活性为6357 U/mL,这也相对高于以往报道。在10 L发酵罐中对大肠杆菌BL21(pET22b-3I)进行发酵时,APL的最终生产效率为282.8 U/mL/h。因此,本研究为碱性果胶酸裂解酶pelNK93I的过表达和制备提供了一种具有成本效益的方法,可用于其工业应用。此外,用于生物精练的pelNK93I(4 U/mL)使棉籽壳去除率和棉织物的径向毛细管效应分别提高了37.63%和47.06%,使其成为绿色纺织技术中一种有前景的酶。