Portal de Periódicos da CAPES

Artigo Acesso aberto Produção Nacional Revisado por pares

BIBTEX RIS

PURIFICATION AND CHARACTERIZATION OF A NEW TRANSGLUTAMINASE FROM STREPTOMYCES SP. ISOLATED IN BRAZILIAN SOIL

2011; Wiley; Volume: 35; Issue: 4 Linguagem: Inglês

10.1111/j.1745-4514.2010.00456.x

1745-4514

Juliana Alves Macedo, Lara Durães Sette, Hélia Harumi Sato,

Proteins in Food Systems

Journal of Food BiochemistryVolume 35, Issue 4 p. 1361-1372 PURIFICATION AND CHARACTERIZATION OF A NEW TRANSGLUTAMINASE FROM STREPTOMYCES SP. ISOLATED IN BRAZILIAN SOIL JULIANA ALVES MACEDO, Corresponding Author JULIANA ALVES MACEDO Food Science Department, Faculty of Food Engineering, Universidade Estadual de Campinas (UNICAMP), PO Box 6121, CEP 13083-862, SP, Brazil TEL: +55-(19)-3521-2175; FAX: +55-(19)-3289-1513; EMAIL: [email protected]Search for more papers by this authorLARA DURÃES SETTE, LARA DURÃES SETTE Brazilian Collection of Environmental and Industrial Microorganisms (CBMAI), Microbial Resource Division, CPQBA/UNICAMP, CEP 13081-970, SP, BrazilSearch for more papers by this authorHÉLIA HARUMI SATO, HÉLIA HARUMI SATO Food Science Department, Faculty of Food Engineering, Universidade Estadual de Campinas (UNICAMP), PO Box 6121, CEP 13083-862, SP, BrazilSearch for more papers by this author JULIANA ALVES MACEDO, Corresponding Author JULIANA ALVES MACEDO Food Science Department, Faculty of Food Engineering, Universidade Estadual de Campinas (UNICAMP), PO Box 6121, CEP 13083-862, SP, Brazil TEL: +55-(19)-3521-2175; FAX: +55-(19)-3289-1513; EMAIL: [email protected]Search for more papers by this authorLARA DURÃES SETTE, LARA DURÃES SETTE Brazilian Collection of Environmental and Industrial Microorganisms (CBMAI), Microbial Resource Division, CPQBA/UNICAMP, CEP 13081-970, SP, BrazilSearch for more papers by this authorHÉLIA HARUMI SATO, HÉLIA HARUMI SATO Food Science Department, Faculty of Food Engineering, Universidade Estadual de Campinas (UNICAMP), PO Box 6121, CEP 13083-862, SP, BrazilSearch for more papers by this author First published: 01 August 2011 https://doi.org/10.1111/j.1745-4514.2010.00456.xCitations: 15 Possible referees: (1) Salvador Mormeneo ([email protected]); (2) Marco Antonio Zachia Ayub ([email protected]). Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Abstract ABSTRACT 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 45 kDa. 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+, Ba2+, Na+, and Co2+, and inhibited by Cu2+ and Hg2+, which suggests a thiol group at its active site. The purified enzyme presented a Km of 6.37 mM and a Vmax of 1.7 U/mL. PRACTICAL APPLICATION Transglutaminase-catalyzed reactions can be used to modify the functional properties of food proteins. Transglutaminase has been used to catalyze the cross-linking of a number of proteins, such as whey proteins, soy proteins, gluten, myosin and actomyosin. The modification of food proteins by transglutaminase may lead to textured products, help to protect lysine residues in food proteins from various chemical reactions, encapsulate lipids and/or lipid-soluble materials, form heat- and water-resistant films, avoid heat treatment in gelation processes, improve elasticity and water holding capacity, modify solubility and functional properties and produce food proteins of higher nutritional value through the cross-linking of different proteins containing complementary limiting essential amino acids. Due all its potentials, the search for new biotechnological sources of transglutaminase and the study of new enzymes characteristics are essential for the reduction of the enzyme costs and development of new applications for the food industry. REFERENCES ANDO, H., ADACHI, M., UMEDA, K., MATSUURA, A., NONAKA, M., UCHIO, R., TANAKA, H. and MOTOKI, M. 1989. Purification and characteristics of a novel transglutaminase derived from microorganisms. Agric. Biol. Chem. Tokyo 53(10), 2613– 2617. BRADFORD, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantitites of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248– 254. CUI, L., DU, G., ZHANG, D., LIU, H. and CHEN, J. 2007. Purification and characterization of transglutaminase from a newly isolated Streptomyces hygroscopicus. Food Chem. 105, 612– 618. FOLK, J.E. and COLE, P.W. 1966. Mechanism of action of pig liver transglutaminase. J. Biol. Chem. 241(23), 5518– 5525. HERRERA, J.R., IRANZO, M., ELORZA, M.V. and SENTANDREU, S.M. 1995. Involvement of transglutaminase in the formation of covalent cross-links in the cell wall of Candida albicans. Arch. Microbiol. 164, 186– 193. HO, M.-L., LEU, S.-Z., HSIEH, J.-F. and JIANG, S.-T. 2000. Technical approach to simplify the purification method and characterization of microbial transglutaminase produced from Streptoverticillium ladakum. Food Chem. Toxicol. 65(1), 76– 80. KLEIN, J., GUZMAN, E. and KUEHN, G. 1992. Purification and partial characterization of transglutaminase from Physarum polycephalum. J. Bacteriol. 174(8), 2599– 2605. KWAN, S.W. and EASA, A.M. 2003. Comparing physical properties of retort-resistant glucono-δ-lactone tofu treated with commercial transglutaminase enzyme or low levels of glucose. Lebensm-Wiss. Technol. 36, 643– 646. LAEMMLI, U. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680– 685. MACEDO, J.A., SETTE, L.D. and SATO, H.H. 2007. Optimization of medium composition for transglutaminase production by a Brazilian soil Streptomyces sp. Electron. J. Biotechnol. [Online] 10(4) http://www.ejbiotechnology.info/content/vol10/issue4/full/10/index.html (accessed January 22, 2009). ISSN 0717-3458. DOI: 10.2225/vol10-issue4-fulltext-10. MACEDO, J.A., SETTE, L.D. and SATO, H.H. 2008. Optimization Studies for the Production of Microbial Transglutaminase from a newly isolated strain of Streptomyces sp. Food Sci. Biotechnol. 17(5), 904– 911. PASTERNACK, R., DORSCH, S., OTTERBACH, J., ROBENEK, I., WOLF, S. and FUNCHSBAUER, H.-L. 1998. Bacterial pro-transglutaminase from Streptoverticillium mobaraense, purification, characterization and sequence of the zymogen. Eur. J. Biochem. 257, 570– 576. SOARES, L.H.B., ASSMANN, F. and AYUB, M.A.Z. 2003. Purification and properties of a transglutaminase produced by a Bacillus circulans strain isolated from the Amazon environment. Biotechol. Appl. Biochem. 37, 297– 299. SOUZA, C., VOLKEN, F., FLORES, S.H. and AYUB, M.A.Z. 2006. Optimization of medium composition for the production of transglutaminase by Bacillus circulans BL32 using statistical experimental methods. Process Biochem. 41, 1186– 1192. SUZUKI, S., IZAWA, Y., KOBAYASHI, K., ETO, Y., YAMANAKA, S., KUBOTA, K. and YOKOZEKI, K. 2000. Purification and characterization of novel transglutaminase from Bacillus subtilis spores. Biosci. Biotechnol. Biochem. 64, 2344– 2351. WORRATAO, A. and YONGSAWATDIGUL, J. 2005. Purification and characterization of transglutaminase from tropical tilapia (Oreochromis niloticus). Food Chem. 93, 651– 658. YOKOYAMA, K., NIO, N. and KIUCHI, Y. 2004. Properties and application of microbial transglutaminase. Appl. Microbiol. Biotechnol. 46, 447– 454. ZHU, Y., RINZEMA, A., TRAMPER, J. and BOL, J. 1995. Microbial transglutaminase – a review of its production and application in food processing. Appl. Microbiol. Biotechnol. 44, 277– 282. Citing Literature Volume35, Issue4August 2011Pages 1361-1372 ReferencesRelatedInformation