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PURIFICATION AND PROPERTIES OF TRANSGLUTAMINASE FROM STREPTOVERTICILLIUM MOBARAENSE
Author(s) -
LU S.Y.,
ZHOU N.D.,
TIAN Y.P.,
LI H.Z.,
CHEN J.
Publication year - 2003
Publication title -
journal of food biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 47
eISSN - 1745-4514
pISSN - 0145-8884
DOI - 10.1111/j.1745-4514.2003.tb00270.x
Subject(s) - centrifugation , sephadex , chromatography , chemistry , enzyme , size exclusion chromatography , tissue transglutaminase , specific activity , pmsf , ethanol precipitation , homogenization (climate) , column chromatography , enzyme assay , biochemistry , biology , extraction (chemistry) , biodiversity , ecology
Transglutaminase (TGase) was separated from the culture broth of an isolated strain of Streptoverticillium mobaraense. The crude enzyme was prepared by centrifugation, ultrafiltration, precipitation by alcohol, centrifugation and freeze‐drying. The yield after these processes was 65–70%. Then the enzyme was purified to homogeneity by chromatography on CM‐cellulose and Sephadex G‐75 on which the yields were about 70% and 80%, respectively; the purified folds reached 2.5–4.7 and 1.08–2.06, respectively. The molecular weight of this TGase was 39,500–40,100 Da by gel filtration chromatography. Optimum enzyme activity was observed in the pH range of 5.0–7.0, and it was maintained stable at 20–40C. The optimal temperature and pH was 52C and 6.0, respectively. At 1 mM and 5 mM metal ion or inhibitors concentration, TGase activity was strongly inhibited by Zn 2+ and NEM, and not affected obviously by Ba 2+ , Ca 2+ , Co 2+ , Cu 2+ , Fe 2+ , Fe 3+ , Mg 2+ , Mn 2+ , Na + as well as PMSF and EDTA. The effects of these additions on this TGase were compared with those of other microbial TGases.