Characterization of a vanillate non-oxidative decaroxylation gene cluster from streptomyces sp.d7
department or school: Science, Faculty of
department or school: Microbiology and Immunology, Department of
data provider: DSpace
degree campus: UBCV
creator: Chow, Kevin Toshio
issued: 2009-07-28T19:29:01Z
issued: 2000
related degree: Doctor of Philosophy - PhD
degree grantor: University of British Columbia
description: The genetics of non-oxidative decarboxylation of aromatic acids to phenolic compounds are poorly understood in both prokaryotes and eukaryotes. Although such reactions have been observed in numerous microorganisms acting on. a variety of substrates, genetic analyses of these processes have not, to my knowledge, been reported in the literature. Previously, I isolated a streptomycete from soil (Streptomyces sp. D7), which efficiently converts 4-hydroxy-3-methoxybenzoic (vanillic) acid to 2-methoxyphenol (guaiacol). Protein two-dimensional gel electrophoresis revealed that several proteins are synthesized in response to vanillic acid, one of which was characterized by partial amino-terminal sequencing, leading to the cloning of a gene cluster from a genomic lambda phage library of Streptomyces sp. D7. This cluster consists of four open reading frames, vdcA (sequencing in progress), vdcB (602 bp), vdcC (1424 bp) and vdcD (239 bp). Protein sequence comparisons suggest that the product of vdcB (201 aa) is similar to phenylacrylate decarboxylase of yeast; the putative products of vdcC (475 aa) and vdcD (80 aa) are similar to hypothetical proteins of unknown function from various microorganisms, and are found in a similar gene cluster in Bacillus subtilis. VdcA is a putative transcriptional regulatory gene. VdcB, vdcC and vdcD homologues are also clustered, along with putative />-cresol methylhydroxylase and vanillin oxidoreductase genes, on the 184 kb catabolic plasmid pNLl of Sphingomonas aromaticivorans F199. Northern blot analysis revealed the synthesis of a 2.5 kb mRNA transcript, which hybridized strongly to a vdcC gene probe, in vanillic acid-induced cells, suggesting that the cluster is under the control of a single inducible promoter. Expression of the entire vdc gene cluster in Streptomyces lividans 1326, as a heterologous host, resulted in that strain acquiring the ability to decarboxylate vanillic acid to guaiacol non-oxidatively. Both Streptomyces strain D7 and…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/11404
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note: CHARACTERIZATION OF A V A N I L L A T E NON-OXIDATIVE D E C A R B O X Y L A T I O N GENE CLUSTER F R O M STREPTOMYCES SP. D7 by K E V I N TOSHIO CHOW M . S c , The University of British Columbia, 1996 B.Sc., The University of British Columbia, 1992 A THESIS SUBMITTED IN PARTIAL F U L F I L L M E N T OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE F A C U L T Y OF G R A D U A T E STUDIES (Department of Microbiology and Immunology) We accept this thesis as conforming to the required standard THE UNIVERSITY OF BRITISH C O L U M B I A December 1999 © Kevin Toshio Chow, 1999 in presenting this thesis in partial fulfilment of the requirements for an advanced degree at the University of British Columbia, I agree that the Library shall make it freely available for reference and study. I further agree that permission for extensive copying of this thesis for scholarly purposes may be granted by the head of my department or by his or her representatives. It is understood that copying or publication of this thesis for financial gain shall not be allowed without my written permission. Department of A//C&&I'QlOfr/ fflfJ* /MMtf^0Lcl 6rY The University of British Columbia Vancouver, Canada Date DE-6 (2/88) ii ABSTRACT The genetics of non-oxidative decarboxylation of aromatic acids to phenolic compounds are poorly understood in both prokaryotes and eukaryotes. Although such reactions have been observed in numerous microorganisms acting on. a variety of substrates, genetic analyses of these processes have not, to my knowledge, been reported in the literature. Previously, I isolated a streptomycete from soil (Streptomyces sp. D7), which efficiently converts 4-hydroxy-3-methoxybenzoic (vanillic) acid to 2-methoxyphenol (guaiacol). Protein two-dimensional gel electrophoresis revealed that several proteins are synthesized in response to vanillic acid, one of which was characterized by partial amino-terminal sequencing, leading to the cloning of a gene cluster from a gen…
has type: Thesis/Dissertation
date issued: 2000-05
is shown at: 10.14288/1.0089827
language: eng
degree discipline: Microbiology and Immunology
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
scholar level: Graduate
type: Text
identifier URI: http://hdl.handle.net/2429/11404