Purification And Characterization Of A New Transglutaminase From Streptomyces Sp. Isolated In Brazilian Soil

A new microbial transglutaminase (MTGase or MTG, EC 2.3.2.13) from a Streptomyces sp. strain isolated from Brazilian soil samples was purified and characterized. Enzyme purification was fast and simple, consisting of two successive chromatographies on Sephadex G-75 columns with yields of 48 and 17%, respectively. The protein purification was successfully achieved to electrophoretical homogeneity on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The molecular mass of the MTGase was estimated to be about 45kDa. The enzyme exhibited optimal activity in the pH range of 6.0-6.5 and at 35-40C. It was stable over a broad pH range (4.5-8.0) and up to 45C. The purified MTG's activity was independent of Ca +2, but was activated by the presence of K +, Ba 2+, Na +, and Co 2+, and inhibited by Cu 2+ and Hg 2+, which suggests a thiol group at its active site. The purified enzyme presented a K m of 6.37mM and a V max of 1.7U/mL. © 2011 Wiley Periodicals, Inc..35413611372Ando, H., Adachi, M., Umeda, K., Matsuura, A., Nonaka, M., Uchio, R., Tanaka, H., Motoki, M., Purification and characteristics of a novel transglutaminase derived from microorganisms (1989) Agric. Biol. Chem. Tokyo, 53 (10), pp. 2613-2617Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantitites of protein utilizing the principle of protein-dye binding (1976) Anal. Biochem., 72, pp. 248-254Cui, L., Du, G., Zhang, D., Liu, H., Chen, J., Purification and characterization of transglutaminase from a newly isolated Streptomyces hygroscopicus (2007) Food Chem., 105, pp. 612-618Folk, J.E., Cole, P.W., Mechanism of action of pig liver transglutaminase (1966) J. Biol. Chem., 241 (23), pp. 5518-5525Herrera, J.R., Iranzo, M., Elorza, M.V., Sentandreu, S.M., Involvement of transglutaminase in the formation of covalent cross-links in the cell wall of Candida albicans (1995) Arch. Microbiol., 164, pp. 186-193Ho, M.-L., Leu, S.-Z., Hsieh, J.-F., Jiang, S.-T., Technical approach to simplify the purification method and characterization of microbial transglutaminase produced from Streptoverticillium ladakum (2000) Food Chem. Toxicol., 65 (1), pp. 76-80Klein, J., Guzman, E., Kuehn, G., Purification and partial characterization of transglutaminase from Physarum polycephalum (1992) J. Bacteriol., 174 (8), pp. 2599-2605Kwan, S.W., Easa, A.M., Comparing physical properties of retort-resistant glucono-δ-lactone tofu treated with commercial transglutaminase enzyme or low levels of glucose (2003) Lebensm-Wiss. Technol., 36, pp. 643-646Laemmli, U., Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature, 227, pp. 680-685Macedo, J.A., Sette, L.D., Sato, H.H., Optimization of medium composition for transglutaminase production by a Brazilian soil Streptomyces sp (2007) Electron. J. Biotechnol. [Online], 10 (4). , (accessed January 22, 2009). ISSN 0717-3458. DOI: 10.2225/vol10-issue4-fulltext-10Macedo, J.A., Sette, L.D., Sato, H.H., Optimization Studies for the Production of Microbial Transglutaminase from a newly isolated strain of Streptomyces sp (2008) Food Sci. Biotechnol., 17 (5), pp. 904-911Pasternack, R., Dorsch, S., Otterbach, J., Robenek, I., Wolf, S., Funchsbauer, H.-L., Bacterial pro-transglutaminase from Streptoverticillium mobaraense, purification, characterization and sequence of the zymogen (1998) Eur. J. Biochem., 257, pp. 570-576Soares, L.H.B., Assmann, F., Ayub, M.A.Z., Purification and properties of a transglutaminase produced by a Bacillus circulans strain isolated from the Amazon environment (2003) Biotechol. Appl. Biochem., 37, pp. 297-299Souza, C., Volken, F., Flores, S.H., Ayub, M.A.Z., Optimization of medium composition for the production of transglutaminase by Bacillus circulans BL32 using statistical experimental methods (2006) Process Biochem., 41, pp. 1186-1192Suzuki, S., Izawa, Y., Kobayashi, K., Eto, Y., Yamanaka, S., Kubota, K., Yokozeki, K., Purification and characterization of novel transglutaminase from Bacillus subtilis spores (2000) Biosci. Biotechnol. Biochem., 64, pp. 2344-2351Worratao, A., Yongsawatdigul, J., Purification and characterization of transglutaminase from tropical tilapia (Oreochromis niloticus) (2005) Food Chem., 93, pp. 651-658Yokoyama, K., Nio, N., Kiuchi, Y., Properties and application of microbial transglutaminase (2004) Appl. Microbiol. Biotechnol., 46, pp. 447-454Zhu, Y., Rinzema, A., Tramper, J., Bol, J., Microbial transglutaminase - a review of its production and application in food processing (1995) Appl. Microbiol. Biotechnol., 44, pp. 277-28