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Microbial production of l -glutamate and l -glutamine by recombinant Corynebacterium glutamicum harboring Vitreoscilla hemoglobin gene vgb

Vitreoscilla hemoglobin (VHb) gene vgb equipped with a native promoter Pvgb or a tac promoter Ptac was introduced into Corynebacterium glutamicum ATCC14067, respectively. Ptac was proven to be more suitable for expressing VHb protein in higher concentration in both Escherichia coli and C. glutamicum... Full description

Journal Title: Applied Microbiology and Biotechnology 2008, Vol.77(6), pp.1297-1304
Main Author: Liu, Qian
Other Authors: Zhang, Jiao , Wei, Xiao-Xing , Ouyang, Shao-Ping , Wu, Qiong , Chen, Guo-Qiang
Format: Electronic Article Electronic Article
Language: English
Subjects:
vgb
ID: ISSN: 0175-7598 ; E-ISSN: 1432-0614 ; DOI: 10.1007/s00253-007-1254-8
Link: http://dx.doi.org/10.1007/s00253-007-1254-8
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recordid: springer_jour10.1007/s00253-007-1254-8
title: Microbial production of l -glutamate and l -glutamine by recombinant Corynebacterium glutamicum harboring Vitreoscilla hemoglobin gene vgb
format: Article
creator:
  • Liu, Qian
  • Zhang, Jiao
  • Wei, Xiao-Xing
  • Ouyang, Shao-Ping
  • Wu, Qiong
  • Chen, Guo-Qiang
subjects:
  • Corynebacterium glutamicum
  • -glutamate
  • -glutamine
  • hemoglobin
  • vgb
ispartof: Applied Microbiology and Biotechnology, 2008, Vol.77(6), pp.1297-1304
description: Vitreoscilla hemoglobin (VHb) gene vgb equipped with a native promoter Pvgb or a tac promoter Ptac was introduced into Corynebacterium glutamicum ATCC14067, respectively. Ptac was proven to be more suitable for expressing VHb protein in higher concentration in both Escherichia coli and C. glutamicum strains compared with the native vgb promoter Pvgb. VHb-expressing C. glutamicum exhibited higher oxygen uptake rate and enhanced cell growth. Recombinant C. glutamicum harboring vgb gene equipped with Ptac promoter produced 23% more l -glutamate in shake-flask culture and grew to 30% more cell density and formed 22% more l -glutamate in fermentor studies compared with the wild-type strain. When a site-directed mutagenesis in which Tyr405 was replaced by a phenylalanine residue (Y405F) was performed on glutamine synthesis gene, recombinant C. glutamicum overexpressing the mutated gene glnA ′ was able to produce l -glutamine effectively. Co-expression of vgb and glnA ′ genes in C. glutamicum produced 17 g/l l -glutamine in shake flask culture, approximately 30% more than that produced by the recombinant harboring only glnA ′ gene. In fermentor cultivation, the recombinant yielded 25% more cells and produced 40.5 g/l l -glutamine. In this study, it was clearly demonstrated that VHb significantly enhanced cell growth, l -glutamate, and l -glutamine production by recombinant C. glutamicum .
language: eng
source:
identifier: ISSN: 0175-7598 ; E-ISSN: 1432-0614 ; DOI: 10.1007/s00253-007-1254-8
fulltext: fulltext
issn:
  • 1432-0614
  • 14320614
  • 0175-7598
  • 01757598
url: Link


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titleMicrobial production of l -glutamate and l -glutamine by recombinant Corynebacterium glutamicum harboring Vitreoscilla hemoglobin gene vgb
creatorLiu, Qian ; Zhang, Jiao ; Wei, Xiao-Xing ; Ouyang, Shao-Ping ; Wu, Qiong ; Chen, Guo-Qiang
ispartofApplied Microbiology and Biotechnology, 2008, Vol.77(6), pp.1297-1304
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subjectCorynebacterium glutamicum ; -glutamate ; -glutamine ; hemoglobin ; vgb
descriptionVitreoscilla hemoglobin (VHb) gene vgb equipped with a native promoter Pvgb or a tac promoter Ptac was introduced into Corynebacterium glutamicum ATCC14067, respectively. Ptac was proven to be more suitable for expressing VHb protein in higher concentration in both Escherichia coli and C. glutamicum strains compared with the native vgb promoter Pvgb. VHb-expressing C. glutamicum exhibited higher oxygen uptake rate and enhanced cell growth. Recombinant C. glutamicum harboring vgb gene equipped with Ptac promoter produced 23% more l -glutamate in shake-flask culture and grew to 30% more cell density and formed 22% more l -glutamate in fermentor studies compared with the wild-type strain. When a site-directed mutagenesis in which Tyr405 was replaced by a phenylalanine residue (Y405F) was performed on glutamine synthesis gene, recombinant C. glutamicum overexpressing the mutated gene glnA ′ was able to produce l -glutamine effectively. Co-expression of vgb and glnA ′ genes in C. glutamicum produced 17 g/l l -glutamine in shake flask culture, approximately 30% more than that produced by the recombinant harboring only glnA ′ gene. In fermentor cultivation, the recombinant yielded 25% more cells and produced 40.5 g/l l -glutamine. In this study, it was clearly demonstrated that VHb significantly enhanced cell growth, l -glutamate, and l -glutamine production by recombinant C. glutamicum .
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titleMicrobial production of l -glutamate and l -glutamine by recombinant Corynebacterium glutamicum harboring Vitreoscilla hemoglobin gene vgb
descriptionVitreoscilla hemoglobin (VHb) gene vgb equipped with a native promoter Pvgb or a tac promoter Ptac was introduced into Corynebacterium glutamicum ATCC14067, respectively. Ptac was proven to be more suitable for expressing VHb protein in higher concentration in both Escherichia coli and C. glutamicum strains compared with the native vgb promoter Pvgb. VHb-expressing C. glutamicum exhibited higher oxygen uptake rate and enhanced cell growth. Recombinant C. glutamicum harboring vgb gene equipped with Ptac promoter produced 23% more l -glutamate in shake-flask culture and grew to 30% more cell density and formed 22% more l -glutamate in fermentor studies compared with the wild-type strain. When a site-directed mutagenesis in which Tyr405 was replaced by a phenylalanine residue (Y405F) was performed on glutamine synthesis gene, recombinant C. glutamicum overexpressing the mutated gene glnA ′ was able to produce l -glutamine effectively. Co-expression of vgb and glnA ′ genes in C. glutamicum produced 17 g/l l -glutamine in shake flask culture, approximately 30% more than that produced by the recombinant harboring only glnA ′ gene. In fermentor cultivation, the recombinant yielded 25% more cells and produced 40.5 g/l l -glutamine. In this study, it was clearly demonstrated that VHb significantly enhanced cell growth, l -glutamate, and l -glutamine production by recombinant C. glutamicum .
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titleMicrobial production of l -glutamate and l -glutamine by recombinant Corynebacterium glutamicum harboring Vitreoscilla hemoglobin gene vgb
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abstractVitreoscilla hemoglobin (VHb) gene vgb equipped with a native promoter Pvgb or a tac promoter Ptac was introduced into Corynebacterium glutamicum ATCC14067, respectively. Ptac was proven to be more suitable for expressing VHb protein in higher concentration in both Escherichia coli and C. glutamicum strains compared with the native vgb promoter Pvgb. VHb-expressing C. glutamicum exhibited higher oxygen uptake rate and enhanced cell growth. Recombinant C. glutamicum harboring vgb gene equipped with Ptac promoter produced 23% more l -glutamate in shake-flask culture and grew to 30% more cell density and formed 22% more l -glutamate in fermentor studies compared with the wild-type strain. When a site-directed mutagenesis in which Tyr405 was replaced by a phenylalanine residue (Y405F) was performed on glutamine synthesis gene, recombinant C. glutamicum overexpressing the mutated gene glnA ′ was able to produce l -glutamine effectively. Co-expression of vgb and glnA ′ genes in C. glutamicum produced 17 g/l l -glutamine in shake flask culture, approximately 30% more than that produced by the recombinant harboring only glnA ′ gene. In fermentor cultivation, the recombinant yielded 25% more cells and produced 40.5 g/l l -glutamine. In this study, it was clearly demonstrated that VHb significantly enhanced cell growth, l -glutamate, and l -glutamine production by recombinant C. glutamicum .
copBerlin/Heidelberg
pubSpringer-Verlag
doi10.1007/s00253-007-1254-8
pages1297-1304
date2008-01