title: Cloning and identification of a novel tyrosinase and its overexpression in Streptomyces kathirae SC-1 for enhancing melanin production.
abstract: A 30-kDa novel tyrosinase was purified to homogeneity. The Km for L-Dopa and L-tyrosine were determined as 0.42 and 0.25 mM. The 1231 bp (base pair) melC gene and its 167 bp promoter Pskmel were obtained by thermal asymmetric interlaced polymerase chain reaction based on the amino acids fragment obtained from MS results of the purified enzyme. The protein sequence of tyrosinase shows maximum identity (84%) to tyrosinase from Streptomyces galbus. The melC was introduced into S. kathirae. The melanin production and the transcriptional level of melC in recombinant S. kathirae [pIJPskmelmelC] were about 2.1-fold and 2-fold higher than the wild-type strain, respectively. The melanin concentration was maximized at 28.8 g L(-1).
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province 214122, P. R. China The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, P. R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province 214122, P. R. China The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, P. R. China raozhm@jiangnan.edu.cn.
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Rao
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province 214122, P. R. China The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, P. R. China.
authors.forename: Taowei
authors.initials: T
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province 214122, P. R. China The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, P. R. China.
authors.forename: Zaiwei
authors.initials: Z
authors.lastname: Man
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province 214122, P. R. China The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, P. R. China.
authors.forename: Meijuan
authors.initials: M
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province 214122, P. R. China The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, P. R. China.
authors.forename: Xian
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Shang-Tian
authors.initials: ST
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D008543:Melanins / Q000096:biosynthesis*; D018832:Molecular Chaperones / Q000235:genetics; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D009876:Operon; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000378:metabolism; D017421:Sequence Analysis; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; C082891:MelC1 protein, Streptomyces; D008543:Melanins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D015534:Trans-Activators; D014442:Monophenol Monooxygenase
keywords: Streptomyces kathirae; enzyme properties; gene cloning; gene expression; melanin production; tyrosinase