COMPLETE NUCLEOTIDE-SEQUENCE AND POLYPEPTIDE ANALYSIS OF MULTICOMPONENT PHENOL HYDROXYLASE FROM PSEUDOMONAS SP STRAIN-CF600

被引:188
作者
NORDLUND, I [1 ]
POWLOWSKI, J [1 ]
SHINGLER, V [1 ]
机构
[1] UMEA UNIV,APPL CELL & MOLEC BIOL UNIT,S-90187 UMEA,SWEDEN
关键词
D O I
10.1128/jb.172.12.6826-6833.1990
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Pseudomonas sp. strain CF600 metabolizes phenol and some of its methylated derivatives via a plasmid-encoded phenol hydroxylase and meta-cleavage pathway. The genes encoding the multicomponent phenol hydroxylase of this strain are located within a 5.5-kb SacI-NruI fragment. We report the nucleotide sequence and the polypeptide products of this 5.5-kb region. A combination of deletion analysis, expression of subfragments in tac expression vectors, and identification of polypeptide products in maxicells was used to demonstrate that the polypeptides observed are produced from the six open reading frames identified in the sequence. Expression of phenol hydroxylase activity in a laboratory Pseudomonas strain allows growth on phenol, owing to expression of this enzyme and the chromosomally encoded ortho-cleavage pathway. This system, in conjunction with six plasmids that each expressed all but one of the polypeptides, was used to demonstrate that all six polypeptides are required for growth on phenol.
引用
收藏
页码:6826 / 6833
页数:8
相关论文
共 24 条
[1]  
Ballou D. P., 1982, FLAVINS FLAVOPROTEIN, P301
[2]  
BARTILSON M, 1990, MOL GEN GENET, V220, P294
[3]   NUCLEOTIDE-SEQUENCE AND EXPRESSION OF THE CATECHOL 2,3-DIOXYGENASE-ENCODING GENE OF PHENOL-CATABOLIZING PSEUDOMONAS CF600 [J].
BARTILSON, M ;
SHINGLER, V .
GENE, 1989, 85 (01) :233-238
[4]  
DAGLEY S, 1986, BACTERIA, V10, P527
[5]   THE XYLABC PROMOTER FROM THE PSEUDOMONAS-PUTIDA TOL PLASMID IS ACTIVATED BY NITROGEN REGULATORY GENES IN ESCHERICHIA-COLI [J].
DIXON, R .
MOLECULAR AND GENERAL GENETICS, 1986, 203 (01) :129-136
[6]  
EBRIGHT RH, 1986, PROTEIN STRUCTURE FO, P207
[7]  
Frantz B., 1986, BACTERIA, V10, P295
[8]   MOLECULAR-CLONING OF THE PLASMID RP4 PRIMASE REGION IN A MULTI-HOST-RANGE TACP EXPRESSION VECTOR [J].
FURSTE, JP ;
PANSEGRAU, W ;
FRANK, R ;
BLOCKER, H ;
SCHOLZ, P ;
BAGDASARIAN, M ;
LANKA, E .
GENE, 1986, 48 (01) :119-131
[9]  
Gibson D. T., 1984, MICROBIAL DEGRADATIO, V13
[10]  
Hanahan D., 1985, DNA CLONING PRACTICA, P109