title: Laccase- and peroxidase-free tyrosinase production by isolated microbial strain.
abstract: Laccase- and peroxidase-free tyrosinase has commercial importance in the production of L-3, 4-dihydroxyphenylalanine (L-DOPA), which is mainly used in the treatment of Parkinson's disease. In the present study, isolation of an actinomycetes microbial strain capable of producing only tyrosinase is reported. Among all soil isolates, three individual colonies revealed black color around the colony in the presence of tyrosine. Further screening for laccase and peroxidase activities using syringaldazine denoted that one of the isolates, designated as RSP-T1, is laccase and peroxidase negative and produces only tyrosinase. The microbe was authenticated as Streptomyces antibioticus based on 16S ribotyping. Effective growth of this isolate was noticed with the use of medium (pH 5.5) containing casein acid hydrolysate (10.0 g/l), K(2)HPO(4) (5.0 g/l), MgSO(4) (0.25 g/l), L-tyrosine (1.0 g/l), and agar (15 g/l). The scanning electron micrograph depicted that the microbe is highly branched and filamentous in nature. The enzyme production was positively regulated in the presence of copper sulfate. The impact of different fermentation parameters on tyrosinase production depicted that the maximized enzyme titer values were observed when this isolate was grown at 6.5 pH and at 30 degrees C temperature under agitated conditions (220 rpm). Among all the studied physiological parameters, agitation played a significant role on tyrosinase production. Upon optimization of the parameters, the yield of tyrosinase was improved more than 100% compared with the initial yield.
authors.affiliation: Department of Biotechnology, Acharya Nagarjuna University, Guntur 522 510, A.P., India. krssrao@yahoo.com
authors.forename: K R S
authors.initials: KR
authors.lastname: Sambasiva Rao
authors.forename: N K
authors.initials: NK
authors.lastname: Tripathy
authors.forename: Y
authors.initials: Y
authors.lastname: Mahalaxmi
authors.forename: R S
authors.initials: RS
authors.lastname: Prakasham
mesh_terms: D016000:Cluster Analysis; D019327:Copper Sulfate / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D020536:Enzyme Activators / Q000378:metabolism; D006835:Hydrazones / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D042845:Laccase / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D009195:Peroxidase / Q000378:metabolism; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D021521:Ribotyping; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013303:Streptomyces antibioticus / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D013696:Temperature; D014443:Tyrosine / Q000378:metabolism
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D020536:Enzyme Activators; D006835:Hydrazones; D012336:RNA, Ribosomal, 16S; C009370:syringaldazine; D014443:Tyrosine; D042845:Laccase; D009195:Peroxidase; D014442:Monophenol Monooxygenase; D019327:Copper Sulfate