Trypsin is a serine trypsin which is widely used in the leather making, food processing, biochemical testing, and pharmaceutical industries. This study aimed to obtain novel trypsin genes from different sources, heterologously expressed in Komagataella phaffii and characterized their enzymatic properties. Through gene mining strategy, potential trypsin genes were obtained from human, bovine, shrimp and Streptomyces fradiae and successfully expressed in the K. phaffii GS115 through pPIC9k vector. After purification and activation, the activities of human-derived trypsin (Homo sapiens trypsin, HST), bovine-derived trypsin (Bos taurus trypsin, BTT), shrimp-derived trypsin (Penaeus vannamei trypsin, PVT) and S. fradiae-derived trypsin (SFT) were 18.6, 12.3, 44.7, and 13.8 U/mL, respectively. The optimum temperatures of the above four types of trypsin were 20, 35, 40, and 45 ℃ while their optimum pH values were 9, 8.5, 8.5, and 9, respectively. Among four types of trypsin, SFT showed the best stability at alkaline environment since it was relatively stable at pH 9-11.
HE Linman
,
NIU Chengtuo
,
LIU Guozheng
,
ZHENG Feiyun
,
LIU Chunfeng
,
WANG Jinjing
,
LI Qi
. Gene mining, heterologous expression and biochemical characterization of trypsin from different sources in Komagataella phaffii GS115[J]. Food and Fermentation Industries, 2024
, 50(2)
: 100
-105
.
DOI: 10.13995/j.cnki.11-1802/ts.035329
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