Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR

• Published in: Electronic Journal of Biotechnology Vol. 39; pp. 91 - 97
• Main Authors: Huang, Lin, Zheng, Dong, Zhao, Yatong, Ma, Jieying, Li, Yanzhen, Xu, Zehua, Shan, Mengying, Shao, Shulin, Guo, Qingwen, Zhang, Jie, Lu, Fuping, Liu, Yihan
• Format: Journal Article
• Language: English
• Published: Elsevier España, S.L.U 01.05.2019; Elsevier
• Subjects: Alkali stability; Bacterial lipases; Biocatalysts; Detergent industry; Directed evolution; Enzymatic detergent; Error-prone PCR; Industrial enzymes; Penicillium cyclopium lipase; Enzymatic detergent; Directed evolution; Detergent industry; Bacterial lipases; Industrial enzymes; Error-prone PCR; Penicillium cyclopium lipase; Biocatalysts; Alkali stability
• ISSN: 0717-3458; 0717-3458
• DOI: 10.1016/j.ejbt.2019.04.002

Lipases are extensively exploited in lots of industrial fields; cold-adapted lipases with alkali-resistance are especially desired in detergent industry. Penicillium cyclopium lipase I (PCL) might be suitable for applications of detergent industry due to its high catalytic efficiency at low temperature and relatively good alkali stability. In this study, to better meet the requirements, the alkali stability of PCL was further improved via directed evolution with error-prone PCR. The mutant PCL (N157F) with an improved alkali stability was selected based on a high-throughput activity assay. After incubating at pH 11.0 for 120 min, N157F retained 70% of its initial activity, which was 23% higher than that of wild type PCL. Combined with the three-dimensional structure analysis, N157F exhibited an improved alkali stability under the high pH condition due to the interactions of hydrophilicity and β-strand propensity. This work provided the theoretical foundation and preliminary data for improving alkali stability of PCL to meet the industrial requirements, which is also beneficial to improving alkali-tolerance ability of other industrial enzymes via molecular modification. How to cite: Huang L, Zheng D, Zhao Y, et al. Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR. Electron J Biotechnol 2019;39. https://doi.org/10.1016/j.ejbt.2019.04.002