Improved production of l-lysine by disruption of stationary phase-specific rmf gene in Escherichia coli

Imaizumi, Akira; Takikawa, Rie; Koseki, Chie; Usuda, Yoshihiro; Yasueda, Hisashi; Kojima, Hiroyuki; Matsui, Kunihiko; Sugimoto, Shinichi

doi:10.1016/j.jbiotec.2004.12.014

Cited by 27 publications

(15 citation statements)

References 17 publications

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“…Much research has reported that E. coli could be genetically modified to produce various amino acids and organic acids by plasmid-mediated homologous recombination (Zhou et al, 2003;Causey et al, 2004;Imaizumi et al, 2005;Zhang et al, 2007). At present, as far as we are aware, gene inactivation of the C. glutamicum genome is mainly carried out by plasmidmediated homologous recombination.…”

Section: Plasmid-mediated Homologous Recombinationmentioning

confidence: 99%

Strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression

Zhang

2016

J. Zhejiang Univ. Sci. B

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Abstract:With the availability of the whole genome sequence of Escherichia coli or Corynebacterium glutamicum, strategies for directed DNA manipulation have developed rapidly. DNA manipulation plays an important role in understanding the function of genes and in constructing novel engineering bacteria according to requirement. DNA manipulation involves modifying the autologous genes and expressing the heterogenous genes. Two alternative approaches, using electroporation linear DNA or recombinant suicide plasmid, allow a wide variety of DNA manipulation. However, the over-expression of the desired gene is generally executed via plasmid-mediation. The current review summarizes the common strategies used for genetically modifying E. coli and C. glutamicum genomes, and discusses the technical problem of multi-layered DNA manipulation. Strategies for gene over-expression via integrating into genome are proposed. This review is intended to be an accessible introduction to DNA manipulation within the bacterial genome for novices and a source of the latest experimental information for experienced investigators.

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Section: Plasmid-mediated Homologous Recombinationmentioning

confidence: 99%

Strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression

Zhang

2016

J. Zhejiang Univ. Sci. B

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“…E.coli DH5α prepared from gene bank Pasteur institute of Iran was grown in Luria Bertani medium (LB) [9] and used for routine transformation. LB contains 10 g/l bacto tryptone, 5 g/l yeast extract and 5 g/l NaCl.…”

Section: Experimental Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

Improvement in the Production of L-Lysine by Overexpression of Aspartokinase (ASK) in <i>C. glutamicum</i> ATCC 21799

Rastegari¹,

Chiani²,

Akbarzadeh³

et al. 2013

Trop. J. Pharm Res

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Purpose: To clone Corynebacterium glutamicum ATCC21799 aspartokinase gene (EC 2.7.2.4) using shuttle expression vector pEKEx2 in order to increase lysine production. Methods: C. glutamicum DNA was extracted and used for amplification of aspartokinase gene (ask) by cloning into an E. coli/C. glutamicum shuttle expression vector, pEKEx2. Initially, the recombinant vector transformed into E. coli DH5α and then into C. glutamicum. Results: Electrophoresis of recombinant protein by SDS-PAGE showed that the molecular weight of the recombinant protein was 42 KD. The induction of recombinant vector by IPTG had an inhibitory effect on cell growth due to over-expression of the cloned gene. The results of lysine assay by Chinard method showed that lysine production increased about two-fold, compared with the parent strain, as a result of increased copy numbers of lysC gene in recombinant strain. Conclusion:A two-fold increase in lysine production was observed by cloning of the ASK gene in C. glutamicum rather than in E. coli, due to the presence of lysine exporter channel which facilitates lysine extraction.

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“…Through comparisons between highly productive and less productive strains of C. glutamicum, NCg10855 and amtA-ocd-soxA operons were selected among the pool of differentially expressed genes and were shown to improve lysine production by as much as 40% [25]. In another similar study, DNA microarray analysis was able to verify the positive effects of knocking down [20] the rmf gene in an L-lysine-producing Escherichia coli strain [26]. In terms of recombinant protein yields, E. coli is the most commonly used system and so identifying genes responsible for stress responses to high cell-densities and/or overexpression of foreign proteins is paramount [27].…”

Section: Engineering Microorganisms (Prokaryotes and Eukaryotes)mentioning

confidence: 99%

Cells by Design: A Mini-Review of Targeting Cell Engineering Using DNA Microarrays

et al. 2008

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Recent studies have demonstrated the utility of DNA microarray technology in engineering cellular properties. For instance, cellular adhesion, the necessity of cells to attach to a surface in order to to proliferate, was examined by comparing two distinct HeLa cell lines. Two genes, one encoding a type II membrane glycosylating sialyltransferase (siat7e) and the other encoding a secreted glycoprotein (lama4), were found to influence adhesion. The expression of siat7e correlated with reduced adhesion, whereas expression of lama4 correlated with increased adhesion, as shown by various assays. In a separate example, a gene encoding a mitochondrial assembly protein (cox15) and a gene encoding a kinase (cdkl3), were found to influence cellular growth. Enhanced expression of either gene resulted in slightly higher specific growth rates and higher maximum cell densities for HeLa, HEK-293, and CHO cell lines. Another investigated property was the adaptation of HEK-293 cells to serum-free media. The genes egr1 and gas6, both with anti-apoptotic properties, were identified as potentially improving adaptability by impacting viability at low serum levels. In trying to control apoptosis, researchers found that by altering the expression levels of four genes faim, fadd, alg-2, and requiem, apoptotic response could be altered. In the present work, these and related studies in microorganisms (prokaryote and eukaryote) are examined in greater detail focusing on the approach of using DNA microarrays to direct cellular behavior by targeting select genes.

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Improved production of l-lysine by disruption of stationary phase-specific rmf gene in Escherichia coli

Cited by 27 publications

References 17 publications

Strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression

Strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression

Improvement in the Production of L-Lysine by Overexpression of Aspartokinase (ASK) in <i>C. glutamicum</i> ATCC 21799

Cells by Design: A Mini-Review of Targeting Cell Engineering Using DNA Microarrays

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