Cover Image

In vitro reconstitution of the catabolic reactions catalyzed by PcaHG, PcaB, and PcaL: the protocatechuate branch of the beta-ketoadipate pathway in Rhodococcus jostii RHA1

The ß-ketoadipate pathway is a major pathway involved in the catabolism of the aromatic compounds in microbes. The recent progress in genome sequencing has led to a rapid accumulation of genes from the ß-ketoadipate pathway in the available genetic database, yet the functions of these genes remain u...

Full description

Saved in:
Bibliographic Details
Main Authors: Yamanashi, Tomoya, Kim, Seungyoung, Hara, Hirofumi, Funa, Nobutaka
Format: Article
Published: Japan Society for Bioscience Biotechnology and Agrochemistry 2015
Subjects:
T Technology (General)
Online Access:http://eprints.utm.my/id/eprint/55814/
http://dx.doi.org/10.1080/09168451.2014.993915
Tags: Add Tag
No Tags, Be the first to tag this record!
id my.utm.55814
record_format eprints
spelling my.utm.558142017-02-15T01:08:56Z http://eprints.utm.my/id/eprint/55814/ In vitro reconstitution of the catabolic reactions catalyzed by PcaHG, PcaB, and PcaL: the protocatechuate branch of the beta-ketoadipate pathway in Rhodococcus jostii RHA1 Yamanashi, Tomoya Kim, Seungyoung Hara, Hirofumi Funa, Nobutaka T Technology (General) The ß-ketoadipate pathway is a major pathway involved in the catabolism of the aromatic compounds in microbes. The recent progress in genome sequencing has led to a rapid accumulation of genes from the ß-ketoadipate pathway in the available genetic database, yet the functions of these genes remain uncharacterized. In this study, the protocatechuate branch of the ß-ketoadipate pathway of Rhodococcus jostii was reconstituted in vitro. Analysis of the reaction products of PcaHG, PcaB, and PcaL was achieved by high-performance liquid chromatography. These reaction products, ß-ketoadipate enol-lactone, 3-carboxy-cis,cis-muconate, y-carboxymuconolactone, muconolactone, and ß-ketoadipate, were further characterized using LC-MS and nuclear magnetic resonance. In addition, the in vitro reaction of PcaL, a bidomain protein consisting of y-carboxy-muconolactone decarboxylase and ß-ketoadipate enol-lactone hydrolase activities, was demonstrated for the first time. This work provides a basis for analyzing the catalytic properties of enzymes involved in the growing number of ß-ketoadipate pathways deposited in the genetic database Japan Society for Bioscience Biotechnology and Agrochemistry 2015 Article PeerReviewed Yamanashi, Tomoya and Kim, Seungyoung and Hara, Hirofumi and Funa, Nobutaka (2015) In vitro reconstitution of the catabolic reactions catalyzed by PcaHG, PcaB, and PcaL: the protocatechuate branch of the beta-ketoadipate pathway in Rhodococcus jostii RHA1. Bioscience, Biotechnology and Biochemistry, 79 (5). pp. 830-835. ISSN 0916-8451 http://dx.doi.org/10.1080/09168451.2014.993915 DOI:10.1080/09168451.2014.993915
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
topic T Technology (General)
spellingShingle T Technology (General)
Yamanashi, Tomoya
Kim, Seungyoung
Hara, Hirofumi
Funa, Nobutaka
In vitro reconstitution of the catabolic reactions catalyzed by PcaHG, PcaB, and PcaL: the protocatechuate branch of the beta-ketoadipate pathway in Rhodococcus jostii RHA1
description The ß-ketoadipate pathway is a major pathway involved in the catabolism of the aromatic compounds in microbes. The recent progress in genome sequencing has led to a rapid accumulation of genes from the ß-ketoadipate pathway in the available genetic database, yet the functions of these genes remain uncharacterized. In this study, the protocatechuate branch of the ß-ketoadipate pathway of Rhodococcus jostii was reconstituted in vitro. Analysis of the reaction products of PcaHG, PcaB, and PcaL was achieved by high-performance liquid chromatography. These reaction products, ß-ketoadipate enol-lactone, 3-carboxy-cis,cis-muconate, y-carboxymuconolactone, muconolactone, and ß-ketoadipate, were further characterized using LC-MS and nuclear magnetic resonance. In addition, the in vitro reaction of PcaL, a bidomain protein consisting of y-carboxy-muconolactone decarboxylase and ß-ketoadipate enol-lactone hydrolase activities, was demonstrated for the first time. This work provides a basis for analyzing the catalytic properties of enzymes involved in the growing number of ß-ketoadipate pathways deposited in the genetic database
format Article
author Yamanashi, Tomoya
Kim, Seungyoung
Hara, Hirofumi
Funa, Nobutaka
author_facet Yamanashi, Tomoya
Kim, Seungyoung
Hara, Hirofumi
Funa, Nobutaka
author_sort Yamanashi, Tomoya
title In vitro reconstitution of the catabolic reactions catalyzed by PcaHG, PcaB, and PcaL: the protocatechuate branch of the beta-ketoadipate pathway in Rhodococcus jostii RHA1
title_short In vitro reconstitution of the catabolic reactions catalyzed by PcaHG, PcaB, and PcaL: the protocatechuate branch of the beta-ketoadipate pathway in Rhodococcus jostii RHA1
title_full In vitro reconstitution of the catabolic reactions catalyzed by PcaHG, PcaB, and PcaL: the protocatechuate branch of the beta-ketoadipate pathway in Rhodococcus jostii RHA1
title_fullStr In vitro reconstitution of the catabolic reactions catalyzed by PcaHG, PcaB, and PcaL: the protocatechuate branch of the beta-ketoadipate pathway in Rhodococcus jostii RHA1
title_full_unstemmed In vitro reconstitution of the catabolic reactions catalyzed by PcaHG, PcaB, and PcaL: the protocatechuate branch of the beta-ketoadipate pathway in Rhodococcus jostii RHA1
title_sort in vitro reconstitution of the catabolic reactions catalyzed by pcahg, pcab, and pcal: the protocatechuate branch of the beta-ketoadipate pathway in rhodococcus jostii rha1
publisher Japan Society for Bioscience Biotechnology and Agrochemistry
publishDate 2015
url http://eprints.utm.my/id/eprint/55814/
http://dx.doi.org/10.1080/09168451.2014.993915
_version_ 1643653909136277504
score 13.211869

Similar Items