title: Production and characterization of tyrosinase activity in Pycnoporus sanguineus CCT-4518 Crude extract.
abstract: Tyrosinase is an enzyme of industrial interest. The production and characterization of tyrosinase from P. sanguineus CCT-4518 were investigated. The selection of inductors, luminosity influence, inoculum size and type of culture medium on the production of tyrosinase and the effect of inhibitors on enzyme activity were performed. Optimum conditions for intracellular tyrosinase production was observed after 2 days using 0.15% L-tyrosine as inducer, in the presence of light, with inoculum size of 10 mycelium discs, using 2% malt extract broth medium, incubated at 30°C, and constant agitation of 150 rpm. Tyrosinase activity was completely inhibited by the addition of 6 mM salicylhydroxamic acid or phenylthiourea, however an inhibition of 4.15% was recorded by the addition of 0.1 mM sodium azide. No inhibition could be detected in case of 0.1 mM phenyl methanesulfonyl fluoride addition. Optimal conditions for intracellular tyrosinase activity using L-dopa as substrate were observed at pH 6.6 and 45°C. Thermal stability studies indicated that the enzyme is stable at 45°C for 15 minutes. Higher temperatures decreased tyrosinase activity. Enzyme production was confirmed by non-denaturing polyacrylamide gel electrophoresis and the protein profile was investigated by denaturing polyacrylamide gel electrophoresis.
authors.affiliation: Laboratório de Enzimologia, Faculdade de Farmácia, Universidade Federal de Goiás , Goiânia, GO , Brasil.
authors.forename: Lívia Teixeira
authors.initials: LT
authors.lastname: Duarte
authors.forename: Joyce Batista
authors.initials: JB
authors.lastname: Tiba
authors.forename: Mariângela Fontes
authors.initials: MF
authors.lastname: Santiago
authors.forename: Telma Alves
authors.initials: TA
authors.lastname: Garcia
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Freitas Bara
keywords: MBTH; Pycnoporus sanguineus; biotechnology; tyrosinase; white-rot fungi
references.citation: J Appl Microbiol. 2006 Feb;100(2):219-32
references.pmid: 16430498
references.citation: J Agric Food Chem. 1999 Feb;47(2):374-8
references.pmid: 10563903
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: FEMS Microbiol Lett. 2000 Feb 15;183(2):301-6
references.pmid: 10675601
references.citation: FEBS Lett. 1990 Jul 2;267(1):99-102
references.pmid: 2365094
references.citation: Bioresour Technol. 2002 Oct;85(1):25-9
references.pmid: 12146638
references.citation: Biochim Biophys Acta. 1995 Feb 22;1247(1):1-11
references.pmid: 7873577
references.citation: Anal Chim Acta. 2006 Nov 24;580(2):271-7
references.pmid: 17723784
references.citation: Appl Microbiol Biotechnol. 2007 May;75(2):311-8
references.pmid: 17216440
references.citation: J Appl Microbiol. 2005;98(2):332-43
references.pmid: 15659188
references.citation: Anal Biochem. 1995 Oct 10;231(1):237-46
references.pmid: 8678307
references.citation: Braz J Microbiol. 2009 Oct;40(4):790-4
references.pmid: 24031426
references.citation: Biotechnol Lett. 2006 May;28(9):633-6
references.pmid: 16642300
references.citation: Phytochemistry. 2004 Mar;65(6):671-8
references.pmid: 15016563
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Biochem Biophys Res Commun. 1995 Jul 26;212(3):834-9
references.pmid: 7626118
references.citation: J Biochem Biophys Methods. 1994 Apr;28(3):173-83
references.pmid: 8064113