文献类型：期刊文献

英文题名：Engineered Folding and Secretion for Expression Improvement of α-Amylase in Komagataella Phaffii

作者：Zhang, Yilun[1];Wang, Xingbin[1];Han, Bei[1];Yao, Chaoying[1];Liu, Qi[1,2];Cai, Menghao[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg, Shanghai, Peoples R China

年份：2025

卷号：4

期号：1

起止页码：3

外文期刊名：FOOD BIOENGINEERING

收录：EI(收录号：20251318107515);WOS:【ESCI(收录号:WOS:001470126500002)】；

基金：This study was supported by the National Key Research and Development Program of China (2022YFC2805102); China Postdoctoral Science Foundation (2022M711146); National Natural Science Foundation of China (32201206).

语种：英文

外文关键词：K. phaffii; molecular chaperone; multicopy; vesicle transport factor; alpha-amylase

摘要：alpha-Amylase is the second most widely produced enzyme globally, with diverse applications in the fields of food, pharmaceutical, bioenergy, papermaking, etc. However, natural alpha-amylase often fails to withstand the extreme conditions encountered in industrial processes, such as low pH and high temperatures. Previous studies identified an alpha-amylase derived from deep-sea sources with resistance to low pH, and subsequent amino acid mutations well enhanced its thermal stability. Nevertheless, the advantageous enzyme mutant exhibited low expression levels in Escherichia coli, highlighting the need for a more suitable expression host. In this study, an engineered industrial host, Komagataella phaffii, was involved for heterologous production of alpha-amylase. High-efficiency signal peptides were screened and multi-copy integrant strains were constructed to achieve a high-yield strain. A total of 31 key chaperones and 11 vesicle transport factors were further investigated to facilitate protein folding and secretion, which resulted in a 3.4-fold increase in alpha-amylase production. Finally, batch fermentation in a 3-L bioreactor achieved a maximum alpha-amylase activity of 2.5 x 104 U/mL. This study demonstrates the development of a high-yield alpha-amylase strain for potential industrial applications, offering valuable insights and strategies for engineering high-producing strains of other industrial enzymes.