High-level expression of a heterologous gene in Escherichia coli in response to carbon-nitrogen source and C/N ratio in a batch bioreactor

Sanjoy K Bhattacharya, Ashok K. Dubey

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Overexpression of the target gene in a recombinant strain of Escherichia coli has been analyzed in view of changes in the carbon-nitrogen source and as a function of C/N ratio. The rate of target gene expression varied over a range of 200%, and the yield coefficient for the target gene product changed over 400% with change in carbon source at 10 g L-1 in M9 medium. With variation in nitrogen source, however, only marginal changes (10-15%) occurred in these parameters of overexpression. Higher concentration, for example 2.5 g L-1, of any nitrogen source adversely affected heterologous expression as rate of target gene expression declined in the range 35%-50% and the yield coefficient of foreign protein decreased between 45% and 60%. A C/N ratio of 15 was found to be optimum for both parameters for overexpression and host specific parameters such as specific growth rate and observed yield coefficient for cellular growth. An analysis with respect to temperature and pH revealed that host and expression parameters were quite susceptible to changes in these factors.

Original languageEnglish
Pages (from-to)151-155
Number of pages5
JournalBiotechnology Progress
Volume13
Issue number2
DOIs
StatePublished - Mar 1 1997
Externally publishedYes

Fingerprint

Bioreactors
bioreactors
carbon nitrogen ratio
Nitrogen
Carbon
Escherichia coli
carbon
nitrogen
Genes
Gene Expression
gene expression
Growth
specific growth rate
genes
Temperature
heterologous gene expression
temperature
Proteins
proteins

ASJC Scopus subject areas

  • Food Science
  • Biotechnology
  • Microbiology

Cite this

@article{b6af6ce6b9f5418f9e249506851c892b,
title = "High-level expression of a heterologous gene in Escherichia coli in response to carbon-nitrogen source and C/N ratio in a batch bioreactor",
abstract = "Overexpression of the target gene in a recombinant strain of Escherichia coli has been analyzed in view of changes in the carbon-nitrogen source and as a function of C/N ratio. The rate of target gene expression varied over a range of 200{\%}, and the yield coefficient for the target gene product changed over 400{\%} with change in carbon source at 10 g L-1 in M9 medium. With variation in nitrogen source, however, only marginal changes (10-15{\%}) occurred in these parameters of overexpression. Higher concentration, for example 2.5 g L-1, of any nitrogen source adversely affected heterologous expression as rate of target gene expression declined in the range 35{\%}-50{\%} and the yield coefficient of foreign protein decreased between 45{\%} and 60{\%}. A C/N ratio of 15 was found to be optimum for both parameters for overexpression and host specific parameters such as specific growth rate and observed yield coefficient for cellular growth. An analysis with respect to temperature and pH revealed that host and expression parameters were quite susceptible to changes in these factors.",
author = "Bhattacharya, {Sanjoy K} and Dubey, {Ashok K.}",
year = "1997",
month = "3",
day = "1",
doi = "10.1021/bp970007t",
language = "English",
volume = "13",
pages = "151--155",
journal = "Biotechnology Progress",
issn = "8756-7938",
publisher = "John Wiley and Sons Ltd",
number = "2",

}

TY - JOUR

T1 - High-level expression of a heterologous gene in Escherichia coli in response to carbon-nitrogen source and C/N ratio in a batch bioreactor

AU - Bhattacharya, Sanjoy K

AU - Dubey, Ashok K.

PY - 1997/3/1

Y1 - 1997/3/1

N2 - Overexpression of the target gene in a recombinant strain of Escherichia coli has been analyzed in view of changes in the carbon-nitrogen source and as a function of C/N ratio. The rate of target gene expression varied over a range of 200%, and the yield coefficient for the target gene product changed over 400% with change in carbon source at 10 g L-1 in M9 medium. With variation in nitrogen source, however, only marginal changes (10-15%) occurred in these parameters of overexpression. Higher concentration, for example 2.5 g L-1, of any nitrogen source adversely affected heterologous expression as rate of target gene expression declined in the range 35%-50% and the yield coefficient of foreign protein decreased between 45% and 60%. A C/N ratio of 15 was found to be optimum for both parameters for overexpression and host specific parameters such as specific growth rate and observed yield coefficient for cellular growth. An analysis with respect to temperature and pH revealed that host and expression parameters were quite susceptible to changes in these factors.

AB - Overexpression of the target gene in a recombinant strain of Escherichia coli has been analyzed in view of changes in the carbon-nitrogen source and as a function of C/N ratio. The rate of target gene expression varied over a range of 200%, and the yield coefficient for the target gene product changed over 400% with change in carbon source at 10 g L-1 in M9 medium. With variation in nitrogen source, however, only marginal changes (10-15%) occurred in these parameters of overexpression. Higher concentration, for example 2.5 g L-1, of any nitrogen source adversely affected heterologous expression as rate of target gene expression declined in the range 35%-50% and the yield coefficient of foreign protein decreased between 45% and 60%. A C/N ratio of 15 was found to be optimum for both parameters for overexpression and host specific parameters such as specific growth rate and observed yield coefficient for cellular growth. An analysis with respect to temperature and pH revealed that host and expression parameters were quite susceptible to changes in these factors.

UR - http://www.scopus.com/inward/record.url?scp=0031104704&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031104704&partnerID=8YFLogxK

U2 - 10.1021/bp970007t

DO - 10.1021/bp970007t

M3 - Article

C2 - 9104039

AN - SCOPUS:0031104704

VL - 13

SP - 151

EP - 155

JO - Biotechnology Progress

JF - Biotechnology Progress

SN - 8756-7938

IS - 2

ER -