Use of cloned genes of Pseudomonas TOL plasmid to effect biotransformation of benzoates to cis-dihydrodiols and catechols by Escherichia coli cells

Journal Article · Sun Dec 01 00:00:00 EST 1985 · Appl. Environ. Microbiol.; (United States)

OSTI ID:6301010

Zeyer, J; Lehrbach, P R; Timmis, K N

DNA fragments containing the xylD and xylL genes, which specify the broad-specificity enzymes toluate-1,2-dioxygenase and 3,5-cyclohexadiene-1,2-diol-1-carboxylic acid dehydrogenase, respectively, of TOL plasmid pWW0-161 of Pseudomonas putida have previously been cloned in the pBR322 vector plasmid. In this study, Escherichia coli cells containing hybrid plasmids carrying the cloned xylD or xylDL genes quantitatively transformed /sup 14/C-ring- and /sup 14/C-carboxy-labeled benzoate to the pathway intermediates 3,5-cyclohexadiene-1,2-diol-1-carboxylic acid (cis-dihydrodiol) and catechol, respectively. Like P. putida cells, E. coli cells containing the xylD gene transformed a variety of chloro- and hydrocarbon-substituted benzoates. The toluate-1,2-dioxygenase produced in E. coli thus exhibited the broad-substrate-specificity properties of the enzyme in P. putida. Turnover rates by the enzymes in these two bacteria are compared.

• Research Organization: Univ. of Geneva, Switzerland
• OSTI ID: 6301010
• Journal Information: Appl. Environ. Microbiol.; (United States), Vol. 50:6
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
BENZOIC ACID
BIODEGRADATION
ESCHERICHIA COLI
GENETIC ENGINEERING
OXIDOREDUCTASES
ENZYME ACTIVITY
CARBON 14 COMPOUNDS
CHLORINE COMPOUNDS
HYDROCARBONS
METABOLISM
PLASMIDS
PSEUDOMONAS
REACTION INTERMEDIATES
TRACER TECHNIQUES
BACTERIA
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CHEMICAL REACTIONS
DECOMPOSITION
ENZYMES
HALOGEN COMPOUNDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MICROORGANISMS
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
550401* - Genetics- Tracer Techniques
550701 - Microbiology- Tracer Techniques