A novel extracytoplasmic phenol oxidase of Streptomyces:: its possible involvement in the onset of morphogenesis

被引:69
作者
Endo, K [1 ]
Hosono, K [1 ]
Beppu, T [1 ]
Ueda, K [1 ]
机构
[1] Nihon Univ, Dept Appl Biol Sci, Fujisawa, Kanagawa 2528510, Japan
来源
MICROBIOLOGY-SGM | 2002年 / 148卷
关键词
laccase; EpoA; melanin; copper; morphological differentiation; Streptomyces griseus;
D O I
10.1099/00221287-148-6-1767
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Exogenous addition of copper stimulates cellular differentiation in Streptomyces spp. Several lines of evidence suggested a parallel correlation between the stimulatory effect of copper and phenol-oxidizing enzyme activities in Streptomyces griseus. Here a novel extracytoplasmic phenol oxidase (EpoA) associated with cellular development of this organism was identified and characterized. EpoA activity, examined by an in-gel stain procedure with NN'-dimethyl-p-phenylenediamine sulfate as a substrate, was repressed by glucose and induced by copper supplied in the medium. The enzyme activity was abolished and markedly reduced in the mutants for A-factor biosynthesis and amfR, respectively, which suggested that the activity of the enzyme depends on those essential regulators for morphogenesis in S. griseus. EpoA protein was purified to homogeneity and the N-terminal amino acid sequence was determined. A homologous sequence identified in the genomic database of Streptomyces coelicolorA3(2) was used as a probe to clone the complete epoA gene of S. griseus. The deduced amino acid sequence of EpoA revealed that the mature protein with a molecular mass of 34 kDa was preceded by a signal peptide consisting of 34 aa, consistent with EpoA being a secreted enzyme. EpoA was predicted to be a laccase-type oxidase by not only the sequence similarity, but its substrate selectivity, oxidizing not tyrosine but dihydroxyphenylalanine (DOPA) to generate melanin pigment. Introduction of epoA on a plasmid partially restored both the EpoA activity and aerial mycelium productivity in an A-factor-deficient mutant. Exogenous supplementation of a substance synthesized by purified EpoA from DOPA stimulated cellular differentiation in S. griseus and several other species. Ultrafiltration indicated that the molecular mass of the putative stimulant synthesized by EpoA is between 500 and 1000 Da.
引用
收藏
页码:1767 / 1776
页数:10
相关论文
共 29 条
[11]  
Hopwood D.A., 1985, GENETIC MANIPULATION
[12]  
Horinouchi S, 1999, CELL-CELL SIGNALING IN BACTERIA, P193
[13]   UNSTABLE GENETIC DETERMINANT OF A-FACTOR BIOSYNTHESIS IN STREPTOMYCIN-PRODUCING ORGANISMS - CLONING AND CHARACTERIZATION [J].
HORINOUCHI, S ;
KUMADA, Y ;
BEPPU, T .
JOURNAL OF BACTERIOLOGY, 1984, 158 (02) :481-487
[14]   CotA of Bacillus subtilis is a copper-dependent laccase [J].
Hullo, MF ;
Moszer, I ;
Danchin, A ;
Martin-Verstraete, I .
JOURNAL OF BACTERIOLOGY, 2001, 183 (18) :5426-5430
[15]  
KEIJER BJ, 2001, J MOL MICROB BIOTECH, V2, P565
[16]  
Maniatis T., 1982, MOL CLONING A LAB MA
[17]   Transcriptional responses in the hemiparasitic plant Triphysaria versicolor to host plant Signals [J].
Matvienko, M ;
Torres, MJ ;
Yoder, JI .
PLANT PHYSIOLOGY, 2001, 127 (01) :272-282
[18]  
Miyadoh S., 1993, ACTINOMYCETOLOGICA, V9, P100, DOI DOI 10.3209/SAJ.7_100
[19]   Copper induction of laccase isoenzymes in the ligninolytic fungus Pleurotus ostreatus [J].
Palmieri, G ;
Giardina, P ;
Bianco, C ;
Fontanella, B ;
Sannia, G .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2000, 66 (03) :920-924
[20]   Marinomonas mediterranea MMB-1 transposon mutagenesis:: Isolation of a multipotent polyphenol oxidase mutant [J].
Solano, F ;
Lucas-Elío, P ;
Fernández, E ;
Sanchez-Amat, A .
JOURNAL OF BACTERIOLOGY, 2000, 182 (13) :3754-3760