Heterologous expression and enzymatic analysis of Streptomyces griseus trypsin in Streptomyces lividans.
- Author:
Tengbo MA
1
;
Zhenmin LING
;
Zhen KANG
;
Jianghua LI
;
Guocheng DU
;
Jian CHEN
Author Information
1. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu, China.
- Publication Type:Journal Article
- MeSH:
Fermentation;
Recombinant Proteins;
biosynthesis;
genetics;
Streptomyces griseus;
enzymology;
Streptomyces lividans;
genetics;
metabolism;
Trypsin;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2013;29(4):466-479
- CountryChina
- Language:Chinese
-
Abstract:
Trypsin as an important serine protease has been widely used in food, pharmaceutical and tanning industries. In this study, we successfully expressed trypsin (cloning from Streptomyces griseus ATCC10137) in Streptomyces lividans TK24 and comparatively investigated its enzymatic properties. Specifically, applying S. griseus ATCC 10137 genome as template, we obtained the sprT gene and sub-cloned it into the expression plasmid pIJ86, generating the recombinant strain S. lividans TK24/pIJ86-sprT. When cultivated in R2YE and SELF, the activity of rSGT reached 9.21 U/mL and 8.61 U/mL, respectively. Meanwhile, the results of the enzymatic analysis showed that rSGT exhibited a higher acid tolerance and a higher specificity to hydrolyze amide bonds compared with bovine trypsin (BT). In addition, Zn2+ and organic solvents up-regulated esterase and amidase of rSGT. Taken together, the results obtained herein provide meaningful information for further modification of rSGT and its industrial application.
