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Overproduction of Bacterial Trypsin in Streptomyces - Optimization for Streptomyces griseus Trypsin Production by Recombinant Streptomyces  

Kim, Jong-Hee (Department of Food and Nutrition, Seoil College)
Hong, Soon-Kwang (Department of Biological Science & Institute of Bioscience and Biotechnology, Myongji University)
Publication Information
Microbiology and Biotechnology Letters / v.36, no.1, 2008 , pp. 28-33 More about this Journal
Abstract
The expression vector (pWHM3-TR1R2) for sprT gene encoding Streptomyces griseus trypsin (SGT) followed by two regulatory genes, sgtR1 and sgtR2, was introduced into Streptomyces lividans TK24 and Streptomyces griseus IFO 13350. Various media with different compositions were used to maximize the productivity of SGT in the recombinant trains. he SGT productivity was best when the transformant of S. lividans TK24 was cultivated in R2YE medium (0.74 unit/mL) at 5 days of cultivation. C5/L (0.66 unit/mL) medium also gave a good productivity, but Livid (0.08 unit/mL) and NDSK (0.06 unit/mL) yielded poor productivities. S. griseus IFO 13350/pWHM3-TR1R2 produced SGT by 1.518 unit/mL (C5/L), 1.284unit/mL (R2YE),0.932 unit/mL (NDSK), and 0.295 unit/mL (Livid) at 7 days of cultivation, which was much higher than those from S. lividans TK24/TR1R2. The SGT protein was purified from the culture broth of S. griseus IFO 13350/pWHM3-TR1R2 in C5/L to homogeneity via ammonium sulfate fractionation, and CM-sepharose and SP-sepharose column chromatographies. The specific activity of purified SGT was 69,252 unit/mg, and the final purification fold and recovery yield were 6.5 and 1.4%, respectively.
Keywords
Streptomyces griseus; bacterial trypsin; sprT; sgtR1; sgtR2;
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