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PMID: 2808302 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Cloning, expression, and regulation of the Pseudomonas cepacia protocatechuate 3,4-dioxygenase genes.

Journal of bacteriology ·Vol. 171 ·No. 11 ·1989-11-00 ·Pages 5907-14

Zylstra GJ, Olsen RH, Ballou DP

Abstract

The genes for the alpha and beta subunits of the enzyme protocatechuate 3,4-dioxygenase (EC 1.13.11.3) were cloned from the Pseudomonas cepacia DBO1 chromosome on a 9.5-kilobase-pair PstI fragment into the broad-host-range cloning vector pRO2317. The resultant clone was able to complement protocatechuate 3,4-dioxugenase mutations in P. cepacia, Pseudomonas aeruginosa, and Pseudomonas putida. Expression studies showed that the genes were constitutively expressed and subject to catabolite repression in the heterologous host. Since the cloned genes exhibited normal induction patterns when present in P. cepacia DBO1, it was concluded that induction was subject to negative control. Regulatory studies with P. cepacia wild-type and mutant strains showed that protocatechuate 3,4-dioxygenase is induced either by protocatechuate or by beta-carboxymuconate. Further studies of P. cepacia DBO1 showed that p-hydroxybenzoate hydroxylase (EC 1.14.13.2), the preceding enzyme in the pathway, is induced by p-hydroxybenzoate and that beta-carboxymuconate lactonizing enzyme, which catalyzes the reaction following protocatechuate 3,4-dioxygenase, is induced by both p-hydroxybenzoate and beta-ketoadipate.

MeSH Terms
Chromosome Mapping Chromosomes, Bacterial Cloning, Molecular Gene Expression Gene Expression Regulation, Bacterial Gene Expression Regulation, Enzymologic Genes, Bacterial Genetic Vectors Genotype Macromolecular Substances Mutation Oxygenases/genetics Protocatechuate-3,4-Dioxygenase/genetics Pseudomonas/enzymology,genetics Restriction Mapping Species Specificity
Chemicals
Macromolecular Substances Oxygenases Protocatechuate-3,4-Dioxygenase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Zylstra G J
Cellular and Molecular Biology Program, University of Michigan Medical School, Ann Arbor 48109-0620.
Olsen R H
Ballou D P
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.171.11.5907-5914.1989
Published
1989-11-00
Pages
5907-14
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC210453
Subset
IM
Grants
NIGMS NIH HHS · GM-20877 · United States
External Links
PubMed source DOI: 10.1128/jb.171.11.5907-5914.1989 PMC Full Text View journal details
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