Functional expression of trypsin from Streptomyces griseus by Pichia pastoris |
| |
Authors: | Zhenmin Ling Tengbo Ma Jianghua Li Guocheng Du Zhen Kang Jian Chen |
| |
Institution: | 1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China 2. School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China 3. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, 214122, China 4. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China
|
| |
Abstract: | In the present study, the genes encoding trypsinogen and active trypsin from Streptomyces griseus were both cloned and expressed in the methylotrophic yeast Pichia pastoris with the α-factor secretion signal under the control of the alcohol oxidase promoter. The mature trypsin was successfully accumulated extracellularly in soluble form with a maximum amidase activity of 6.6?U?ml?1 (batch cultivation with flask cultivation) or 14.4?U?ml?1 (fed-batch cultivation with a 3-l fermentor). In contrast, the recombinant trypsinogen formed inclusion bodies and no activity was detected. Replacement of the trypsin propeptide Ala-Pro-Asn-Pro confirmed that its physiological function was as a repressor of activity. More importantly, our results proved that the propeptide inhibited the activity of trypsinogen after its successful folding. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|