• Korean Journal of Microbiology
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• v.29 no.3
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• pp.149-154
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• 1991

Escherichia coli/Corynebacterium glutamicum shuttle vectors, pECCG1 and pECCG2 were constructed by joining a 3.00 kb cryptic plasmid pCB 1 from C. glutamicum and a 3.94 kb plasmid pACYC 177 from E. coli. By trimming unessential parts and introducing mulitiple cloning site into the plasmid pECCG 1, a plasmid pECCG122(5.1kb) was constructed. All the shuttle vectors were stably maintained in C. glutamicum up to about 40 generations irrespective of kanamycin addition in the medium. Threonine operon (homoserine dehydrogenase/homoserine kinase) and dapA gene (dihydrodipicolinate synthetase) of C. glutamicum were cloned into the plasmid pECCG122, and the resultant plasmids were designated pTN31 and pDHDP19, respectively. They were used to study the effect of cloned foreign gene on the stability of the plasmid pECCG122. Plasmids pTN31 and pDHDP19 were segregated rapidly from C. glutamicum when cultured in the medium without kanamycin. In medium with $50\mu${\g/ml} of kanamycin, their segregation rates were much slower than those in medium without kanamycin, but the danamycin addition didn't guarantee the complete maintenance of the plasmids in C. glutamicum.

• Microbiology and Biotechnology Letters
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• v.26 no.1
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• pp.45-49
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• 1998

Two kinds of Mutants of Corynebacterium glutamicum, which were resistant to branched chain amino acid analogues, were obtained for L-leucine production; C. glutamicum LT26 resistant to 4-azaleucine and $\alpha$-amino-$eta$-hydroxyvaleric acid, and from which C. glutamicum LT3811-70 resistant to DL-4-thiaisoleucine were derived. Accumulation of L-leucine in the culture broths of these mutant strains, C. glutamicum LT26 and LT3811-70, were much higher than those of their parent strains even though they were non-auxotrophic mutants. Enzymatic analyses were performed to measure the activities of $\alpha$-acetohydroxy acid synthase (AHAS) and $\alpha$-isopropylmalate synthase (IPMS), which were the key enzymes for the L-isoleucine, L-valine and L-leucine biosynthetic pathways branching from a common precursor. In C. glutamicum LT26 and LT3811-70, AHAS and IPMS were found to be derepressed and desensitized to L-leucine. In addition, in C. glutamicum LT3811-70, IPMS was further more derepressed by L-leucine and AHAS was more desensitized by L-isoleucine and L-valine compared to its parent strain, C. gIEitamicum LT26.

• Korean Journal of Microbiology
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• v.29 no.4
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• pp.203-208
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• 1991

The dapA-complementing gene (L-2, 3-dihydrodipicolinate synthetase: DHDP synthetase, dapA) has been cloned by using a cosmid genomic bank of Corynebacterium glutamicum JS231 that is a lysine overproducer, AEC (s-(2-aminoethyl)-L-cysteine) resistant mutant. By enzymatic deletion analysis, the DNA region complementing the escherichia coli dapA host could be confined to 4.5kb SalI-generated DNA fragment. This DNA fragment was inserted into the C. glutamicum/E. coli shuttle vector pECCG117 to construct pDHDP5812. The specific activity of DHDP synthetase detected in C. glutamicum JS231/pDHDP5812 was increased about 10 fold above that of C. glutamicum JS231. The addition of leucine during growth did not repress the expressin of dapA, and the enzyme activity was not inhibited by lysine.

• Microbiology and Biotechnology Letters
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• v.38 no.4
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• pp.349-361
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• 2010

In this review, the current knowledge of the carbon metabolism and global carbon regulation in Corynebacterium glutamicum are summarized. C. gluamicum has phosphotransferase system (PTS) for the utilization of sucrose, glucose, and fructose. C. glutamicum does not show any preference for glucose when various sugars or organic acids are present with glucose, and thus cometabolizes glucose with other sugars or organic acids. The molecular mechanism of global carbon regulation such as carbon catabolite repression (CCR) in C. glutamicum is quite different to that in Gram-negative or low-GC Gram-positive bacteria. GlxR (glyoxylate bypass regulator) in C. glutamicum is the cyclic AMP receptor protein (CRP) homologue of E. coli. GlxR has been reported to regulate genes involved in not only glyoxylate bypass, but also central carbon metabolism and CCR including glycolysis, gluconeogenesis, and tricarboxylic acid (TCA) cycle. Therefore, GlxR has been suggested as a global transcriptional regulator for the regulation of diverse physiological processes as well as carbon metabolism. Adenylate cyclase of C. glutamicum is a membrane protein belonging to class III adenylate cyclases, thus it could possibly be a sensor for some external signal, thereby modulating cAMP level in response to environmental stimuli. In addition to GlxR, three additional transcriptional regulators like RamB, RamA, and SugR are also involved in regulating the expression of many genes of carbon metabolism. Finally, recent approaches for constructing new pathways for the utilization of new carbon sources, and strategies for enhancing amino acid production through genetic modification of carbon metabolism or regulatory network are described.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.541-546
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• 2012

The biodegradation of phenol in laboratory-contaminated soil was investigated using the Gram-positive soil bacterium Corynebacterium glutamicum. This study showed that the phenol degradation caused by C. glutamicum was greatly enhanced by the addition of 1% yeast extract. From the toxicity test using Daphnia magna, the soil did not exhibit any hazardous effects after the phenol was removed using C. glutamicum. Additionally, the treatment of the phenol-contaminated soils with C. glutamicum increased various soil amino acid compositions, such as glycine, threonine, isoleucine, alanine, valine, leucine, tyrosine, and phenylalanine. This phenomenon induced an increase in the seed germination rate and the root elongation of Avena sativa (oat). This probably reflects that increased soil amino acid composition due to C. glutamicum treatment strengthens the plant roots. Therefore, the phenol-contaminated soil was effectively converted through increased soil amino acid composition, and additionally, the phenol in the soil environment was biodegraded by C. glutamicum.

• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.31-36
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• 1991

A 7.5 kilobases hybrid plasmid, designated as pCE1301, was constructed by combining Eschurichia cwli plasmid pBELl which carries the kanamycin resistance gene of Tn5 with a cryptic plasmid, pSRl of Corynebacterium glutamicum. pCE1301 was transformed C. glutaicum by PEG-mediated protoplast method and its transformation efficiency was about $3.0\times 10^3$ transformants per $\mu g$ of the hybrid plasmid DNA. The physical map reveals that pCE1301 has single restriction sites for SalI and EcoRl, respectively. 'The kanamycin resistance of pCE1301 was stably maintained in C. glutamicum up to 25 generations and any segregation was not detected. pCI31301 was also introduced into Brevibacterium flavum and E coil, and replicated in those strains. pCE1301 was proved to be useiul in cloning the homoscrine dehydrogenase gene from C. glutamicum.

• Journal of Microbiology and Biotechnology
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• v.26 no.5
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• pp.807-822
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• 2016

Starting as a glutamate producer, Corynebacterium glutamicum has played a variety of roles in the industrial production of amino acids, one of the most important areas of white biotechnology. From shortly after its genome information became available, C. glutamicum has been applied in various production processes for value-added chemicals, fuels, and polymers, as a key organism in industrial biotechnology alongside the surprising progress in systems biology and metabolic engineering. In addition, recent studies have suggested another potential for C. glutamicum as a synthetic biology platform chassis that could move the new era of industrial microbial biotechnology beyond the classical field. Here, we review the recent progress and perspectives in relation to C. glutamicum, which demonstrate it as one of the most promising and valuable workhorses in the field of industrial biotechnology.

• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.999-1009
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• 2006

Physiological, biochemical and genetic studies of Corynebacterium glutamicum, a workhorse of white biotechnology used for amino acid production, led to a waste knowledge mainly about amino acid biosynthetic pathways and the central carbon metabolism of this bacterium. Spurred by the availability of the genome sequence and of genome-based experimental methods such as DNA microarray analysis, research on genetic regulation came into focus. Recent progress on mechanisms of genetic regulation of the carbon, nitrogen, sulfur and phosphorus metabolism in C. glutamicum will be discussed.

• KSBB Journal
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• v.9 no.4
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• pp.372-377
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• 1994

To improve L-lysine yield, pyrimidine base analogue(6-azauracil)-resistant mutants were isolated from Corynebacterium glutamicum KFCC10672 Among them the best producer, C. glutamicum CH0516, was selected and tested for L-lysine production in a $7\ell$ fermentor. It was found that the product yield obtained with C. glutamicum CH0516 was higher than that of the parent strain by 3%. In order to elucidate the gain in productivity with the 6-azauracil-resistant mutant enzymatic kinetic parameters such as aspartokinase(AKase) and aspartate carbamoyltransferase (ATCase) were measured. The Km values of AKase with C. glutamicum KFCC10672 and CH0516 were 200.0 mM and 166.7 mM and those of ATCase were 0.13 mM and 0.27 mM, respectively. However, the specific enzyme activities of AKase of C. glutamlcum KFCC10672 and CH0516 were $3.89{\times}10^{-1}$ units/mg and $4.78{\times}10^{-1}$ units/mg, and those of ATCarse were 2.20 units/mg and 1.84 units/mg, respectively. It appears that some increase in product yield with C. gluramicum CH0516 is likely due to the increased Akase activity and decreased ATCase activity.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.271-275
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• 2017

The expression of Deinococcus radiodurans dr1558 and pprM genes was examined for enhanced lysine production in recombinant Corynebacterium glutamicum. These genes are known to confer high tolerance to pH and osmotic shock in Escherichia coli. D. radiodurans dr1558 and pprM genes were expressed in C. glutamicum by using 6 synthetic promoters of different strengths, to evaluate the effect of expression efficiency on lysine production. Recombinant C. glutamicum expressing DR1558 under the L26 and I64 promoters showed higher lysine production than that expressing DR1558 under other promoters. Similarly, recombinant C. glutamicum expressing PprM under same promoters (L26 and I64) showed a higher increase in lysine production compared to that expressing PprM under other promoters. In the absence of $CaCO_3$ in the medium, the expression of DR1558 or PprM also increased lysine concentration in C. glutamicum depending on the promoter used. Together, these results suggest that genes involved in radiation tolerance in D. radiodurans can be used to enhance production of amino acids and their derivatives.