Characterization of two Streptomyces enzymes that convert ferulic acid to vanillin.

Production of flavors from natural substrates by microbial transformation has become a growing and expanding field of study over the past decades. Vanillin, a major component of vanilla flavor, is a principal flavoring compound used worldwide. Streptomyces sp. strain V-1 is known to be one of the mo...

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Autores principales:	Wenwen Yang, Hongzhi Tang, Jun Ni, Qiulin Wu, Dongliang Hua, Fei Tao, Ping Xu
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/14595daccbb745049b887c11235a9107
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spelling	oai:doaj.org-article:14595daccbb745049b887c11235a91072021-11-18T07:39:30ZCharacterization of two Streptomyces enzymes that convert ferulic acid to vanillin.1932-620310.1371/journal.pone.0067339https://doaj.org/article/14595daccbb745049b887c11235a91072013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23840666/?tool=EBIhttps://doaj.org/toc/1932-6203Production of flavors from natural substrates by microbial transformation has become a growing and expanding field of study over the past decades. Vanillin, a major component of vanilla flavor, is a principal flavoring compound used worldwide. Streptomyces sp. strain V-1 is known to be one of the most promising microbial producers of natural vanillin from ferulic acid. Although identification of the microbial genes involved in the biotransformation of ferulic acid to vanillin has been previously reported, purification and detailed characterization of the corresponding enzymes with important functions have rarely been studied. In this study, we isolated and identified 2 critical genes, fcs and ech, encoding feruloyl-CoA synthetase and enoyl-CoA hydratase/aldolase, respectively, which are involved in the vanillin production from ferulic acid. Both genes were heterologously expressed in Escherichia coli, and the resting cell reactions for converting ferulic acid to vanillin were performed. The corresponding crucial enzymes, Fcs and Ech, were purified for the first time and the enzymatic activity of each purified protein was studied. Furthermore, Fcs was comprehensively characterized, at an optimal pH of 7.0 and temperature of 30°C. Kinetic constants for Fcs revealed the apparent Km, kcat, and Vmax values to be 0.35 mM, 67.7 s(-1), and 78.2 U mg(-1), respectively. The catalytic efficiency (kcat/Km) value of Fcs was 193.4 mM(-1) s(-1) for ferulic acid. The characterization of Fcs and Ech may be helpful for further research in the field of enzymatic engineering and metabolic regulation.Wenwen YangHongzhi TangJun NiQiulin WuDongliang HuaFei TaoPing XuPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 6, p e67339 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Wenwen Yang Hongzhi Tang Jun Ni Qiulin Wu Dongliang Hua Fei Tao Ping Xu Characterization of two Streptomyces enzymes that convert ferulic acid to vanillin.
description	Production of flavors from natural substrates by microbial transformation has become a growing and expanding field of study over the past decades. Vanillin, a major component of vanilla flavor, is a principal flavoring compound used worldwide. Streptomyces sp. strain V-1 is known to be one of the most promising microbial producers of natural vanillin from ferulic acid. Although identification of the microbial genes involved in the biotransformation of ferulic acid to vanillin has been previously reported, purification and detailed characterization of the corresponding enzymes with important functions have rarely been studied. In this study, we isolated and identified 2 critical genes, fcs and ech, encoding feruloyl-CoA synthetase and enoyl-CoA hydratase/aldolase, respectively, which are involved in the vanillin production from ferulic acid. Both genes were heterologously expressed in Escherichia coli, and the resting cell reactions for converting ferulic acid to vanillin were performed. The corresponding crucial enzymes, Fcs and Ech, were purified for the first time and the enzymatic activity of each purified protein was studied. Furthermore, Fcs was comprehensively characterized, at an optimal pH of 7.0 and temperature of 30°C. Kinetic constants for Fcs revealed the apparent Km, kcat, and Vmax values to be 0.35 mM, 67.7 s(-1), and 78.2 U mg(-1), respectively. The catalytic efficiency (kcat/Km) value of Fcs was 193.4 mM(-1) s(-1) for ferulic acid. The characterization of Fcs and Ech may be helpful for further research in the field of enzymatic engineering and metabolic regulation.
format	article
author	Wenwen Yang Hongzhi Tang Jun Ni Qiulin Wu Dongliang Hua Fei Tao Ping Xu
author_facet	Wenwen Yang Hongzhi Tang Jun Ni Qiulin Wu Dongliang Hua Fei Tao Ping Xu
author_sort	Wenwen Yang
title	Characterization of two Streptomyces enzymes that convert ferulic acid to vanillin.
title_short	Characterization of two Streptomyces enzymes that convert ferulic acid to vanillin.
title_full	Characterization of two Streptomyces enzymes that convert ferulic acid to vanillin.
title_fullStr	Characterization of two Streptomyces enzymes that convert ferulic acid to vanillin.
title_full_unstemmed	Characterization of two Streptomyces enzymes that convert ferulic acid to vanillin.
title_sort	characterization of two streptomyces enzymes that convert ferulic acid to vanillin.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/14595daccbb745049b887c11235a9107
work_keys_str_mv	AT wenwenyang characterizationoftwostreptomycesenzymesthatconvertferulicacidtovanillin AT hongzhitang characterizationoftwostreptomycesenzymesthatconvertferulicacidtovanillin AT junni characterizationoftwostreptomycesenzymesthatconvertferulicacidtovanillin AT qiulinwu characterizationoftwostreptomycesenzymesthatconvertferulicacidtovanillin AT donglianghua characterizationoftwostreptomycesenzymesthatconvertferulicacidtovanillin AT feitao characterizationoftwostreptomycesenzymesthatconvertferulicacidtovanillin AT pingxu characterizationoftwostreptomycesenzymesthatconvertferulicacidtovanillin
_version_	1718423079511654400

Characterization of two Streptomyces enzymes that convert ferulic acid to vanillin.

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