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Heterologous Expression and Purification of Zea mays Transglutaminase in Pichia pastoris
Hongbo Li,Yanhua Cui,Lanwei Zhang,Huaxi Yi,Xue Han,Yuehua Jiao,Ming Du,Rongbo Fan,Shuang Zhang 한국식품과학회 2014 Food Science and Biotechnology Vol.23 No.5
Transglutaminases (TGases) are a family ofenzymes that catalyze the cross-linking of proteins and arewidely used in the food industry to improve the texture ofdairy, meat, and bread products. Zea mays transglutaminase(TGZ) is a new type of TGase with a wide potential. TGZwas expressed in the yeast Pichia pastoris under an alcoholoxidase promoter. Maximal expression of recombinantTGZ was achieved by inducing recombinant GS115(pPIC9K-tgz) in BMMY medium using 1.5% methanol for96 h. Secreted TGZ was initially separated using Superdex200 resin and further purified on cation exchange resin. The activity of TGZ following purification was 0.32 U/mgof protein. The polymerization effect of TGZ on caseincatalyzed by recombinant TGZ was slightly lower than theeffect of microbial transglutaminase (MTG). TGZ is a newpotential additive for the food industry.
Lili Zhang,Lanwei Zhang,Huaxi Yi,Ming Du,Yingchun Zhang,Xue Han,Zhen Feng,Jingyan Li,Yuehua Jiao,Yanhe Zhang,Chunfeng Guo 한국화학공학회 2012 Korean Journal of Chemical Engineering Vol.29 No.7
Stress-mediated bioprocess is a strategy designed to enhance biological target productivity. In this study,an attempt was made to enhance transglutaminase (TGase) production by Streptomyces mobaraensis by using different stress conditions including heat shock, alcohols and salt stress. Results showed that the effects of stress on TGase production depended on the type applied. For heat shock, TGase production (1.32 U/ml) was recorded maximum in the culture treated at 48 h post inoculation in water bath at 60 oC for 1 min. For alcohols treatment, the maximum activity of TGase (1.77 and 1.75 U/ml) was obtained when 3% methanol was added into the medium at 0 or 24 h of fermentation. However, a 3.5-fold increased production of TGase (3.8 U/ml) was observed in the medium supplemented with 0.2mol/l MgCl2 compared with the basic medium at the beginning of fermentation. In conclusion, TGase production from S. mobaraensis was improved by heat shock, methanol and salt stress treatments, MgCl2 stress was the most effective.