• 한국동물위생학회지
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• 제40권1호
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• pp.53-59
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• 2017

Foot and mouth disease (FMD) is highly infectious disease of cloven-hoofed animals, particularly problematic in cattle, sheep, pigs and goats for economic reasons. Last FMD outbreak in February, 2017 caused tremendous social and economical impacts. The Korean FMD policy aims to vaccinate intact animals and euthanize and bury infected animals to prevent the disease spread. However, there was a problem that the buried livestock did not decompose after several years. Therefore, the study was purposed to investigate the effect of Corynebacterium glutamicum and Bacillus licheniformis on the degradation of buried cow carcasses and on the soil condition; such as temperature, decomposition course of carcasses, composition of amino acids in the soil around carcasses, and plant root elongation to measure soil conditions. As a result, the composition of amino acids in the soil treated with C. glutamicum and B. licheniformis was generally higher than those in the untreated soil. Plant roots in soil treated with C. glutamicum and B. licheniformis grew longer than in non-treated soil. The results suggested that the toxic effect on a grave land buried with FMD infected livestock is reduced when treated with C. glutamicum and B. licheniformis in regard of odor reduction, promoted decaying process, and soil fertilization.

• Journal of Microbiology and Biotechnology
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• 제4권4호
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• pp.256-263
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• 1994

The aceB gene, encoding for malate synthase, one of the key enzymes of glyoxylate bypass, was isolated from a pMT1-based Corynebacterium glutamicum gene library via complementation of an Escherichia coli aceB mutant on an acetate minimal medium. The aceB gene was closely linked to aceA, separated by 598 base pairs, and transcribed in divergent direction. The aceB expressed a protein product of Mr 83, 000 in Corynebacterium glutamicum which was unusually large compared with those of other malate synthases. A DNA-sequence analysis of the cloned DNA identified an open-reading frame of 2, 217 base pairs which encodes a protein with the molecular weight of 82, 311 comprising 739 aminoo acids. The putative protein product showed only limited amino acid-sequence homology to its counteliparts in other organisms. The N-terminal region of the protein, which shows no apparent homology with the known sequences of other malate synthases, appeared to be responsible for the protein s unusually large size. A potential calciumbinding domain of EF-hand structure found among eukaryotes was detected in the N-terminal region of the deduced protein.

• 한국유기농업학회지
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• 제7권1호
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• pp.105-113
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• 1998

수종의 근권세균이 양액재배 오이의 생장에 미치는 영향을 구명하기 위하여 Azospirillum sp., Rhodopseudomonas sp., Pseudomonas sp., Bacillus sp.와 Corynebacterium glutamicum간의 융합균주 등을 근권처리하였다. 융합균주의 근권처리후 4일에 세균의 밀도변화르르 보면 코코피트 > 암면 > 배양액 등의 순으로 나타났다. 균주의 근권처리에 따른 오이의 생장촉진효과는 암면경보다 코코피트경에서 더 높았으며, 균주의 종류별 오이의 생장촉진효과를 보면 Azospirillum sp. > 광합성세균 $\ge$ 융합균주 Pseudomonas sp. > 대조구 등의 순으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.704-710
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• 2008

Glutamate availability in the argF-argR-proB${\Delta}$ strain of Corynebacterium glutamicum was increased by addition of glutamate to the cell or inactivation of the phosphoenolpyruvate carboxykinase activity and simultaneous overexpression of the pyruvate carboxylase activity to assess its effect on L-ornithine production. When glutamate was increased in an L-ornithine-producing strain, the production of L-ornithine was not changed. This unexpected result indicated that the intracellular concentration and supply of glutamate is not a rate-limiting step for the L-ornithine production in an L-ornithine-producing strain of C. glutamicum. In contrast, overexpression of the L-ornithine biosynthesis genes (argCJBD) resulted in approximately 30% increase of L-ornithine production, from 12.73 to 16.49 mg/g (dry cell weight). These results implied that downstream reactions converting glutamate to L-ornithine, but not the availability of glutamate, is the rate-limiting step for elevating L-ornithine production in the argF-argR-proB${\Delta}$ strain of C. glutamicum.

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.127-131
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• 2010

In this study, Corynebacterium glutamicum and its derived mutants were used to demonstrate the relationship between proline, glutamate, and ornithine. The maximum ornithine production was shown in the culture medium (3,295.0 mg/l) when the cells were cultured with 20 mM proline, and was 15.5 times higher than in the presence of 1 mM proline. However, glutamate, which is known as an intermediate in the process of converting proline to ornithine, did not have any positive effect on ornithine production. This suggests that the conversion of proline to ornithine through glutamate, is not possible in C. glutamicum. Comparative analysis between the wild-type strain, SJC 8043 ($argF^-$, $argR^-$), and SJC 8064 ($argF^-$, $argR^-$, and $ocd^-$), showed that C glutamicum could regulate ornithine production by ornithine cyclodeaminase (Ocd) under proline-supplemented conditions. Therefore, proline directly caused an increase in the endogenous level of ornithine by Ocd, which would be a primary metabolite in the ornithine biosynthesis pathway.

• Journal of Microbiology and Biotechnology
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• 제33권10호
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• pp.1361-1369
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• 2023

Corynebacterium glutamicum (C. glutamicum) has been considered a very important and meaningful industrial microorganism for the production of amino acids worldwide. To produce amino acids, cells require nicotinamide adenine dinucleotide phosphate (NADPH), which is a biological reducing agent. The pentose phosphate pathway (PPP) can supply NADPH in cells via the 6-phosphogluconate dehydrogenase (6PGD) enzyme, which is an oxidoreductase that converts 6-phosphogluconate (6PG) to ribulose 5-phosphate (Ru5P), to produce NADPH. In this study, we identified the crystal structure of 6PGD_apo and 6PGD_NADP from C. glutamicum ATCC 13032 (Cg6PGD) and reported our biological research based on this structure. We identified the substrate binding site and co-factor binding site of Cg6PGD, which are crucial for understanding this enzyme. Based on the findings of our research, Cg6PGD is expected to be used as a NADPH resource in the food industry and as a drug target in the pharmaceutical industry.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.789-795
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• 2004

A promoter-probe shuttle vector pSK1Cat was constructed for the isolation of transcriptional signal sequences from Corynebacterium glutamicum. Besides conferring resistance to kanamycin in Escherichia coli and C. glutamicum, the vector carried a promoterless cat gene to confer resistance to chloramphenicol upon insertion of the appropriate transcriptional signals in the multiple cloning site. By utilizing the vector, a series of transcriptionally active fragments were isolated from the genome of C. glutamicum. The clones, ranging from 200 bp to 1 kb in size, were grouped into 3 classes of strong, medium, and weak, based on the chloramphenicol acetyltransferase (CAT) activity and sensitivity to the chloramphenicol of the clone-carrying C. glutamicum cells. C. glutamicum cells carrying the $P_{19}$ clone, a representative in the strong class, were able to grow on minimal agar plates containing over $40 mg/mell$ chloramphenicol, and showed CAT activity of 10 m㏖/mgㆍmin, performing slightly better than the cells carrying $P_{tac}$ , a strong E. coli promoter. Subcloning analysis of the $P_{19}$ clone identified a 180 bp intergenic fragment ($P_{180}$), which was located upstream of a gene encoding a hypothetical membrane protein. The expression conferred by $P_{180}$ was not affected by either the kinds of carbon sources or changes in temperature. These properties make the $P_{180}$ clone useful for the deregulated expression of biosynthetic genes in C. glutamicum during amino acid fermentation.

• 미생물학회지
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• 제41권4호
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• pp.300-305
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• 2005

Corynebacterium glutamicum에서 promoter-probe vector인 pSK1Cat을 이용해 분리된 프로모터를 함유하는 단편들 중 가장 높은 활성을 나타낸 $P_{19}$ 단편에 대한 심도 있는 분석을 수행하였다. Subcloning을 실시하여 프로모터 활성을 지닌 DNA 영역을 180 bp로 압축할 수 있었고 $(P_{180})$, 이를 C. glutamicum의 균주개량 측면에서 그 활용성을 분석하였다. C. glutamicum에서 메치오닌 생합성에 관여하는metX유전자의 메치오닌에 의한 repression을 해제시키기 위하여 metX유전자의 promoter를 $P_{180}$ promoter로 교체하였고 $(P_{180}-metX)$, $P_{180}-metX$를 C. glutamicum에 도입하여 발현되는 homoserine acetyltransferase 활성을 다양한 성장조건에서 측정하였다. MB 영양배지에서 배양하는 경 우 $P_{180}-metX$를 함유는 균주는 wild type보다 약 24배 높은 homoserine acetyltransferase 활성을 나타내었다. Tac 프로모터에 연계하는 경우 $(P_{tac}-metX)$, 약 13배의 활성 증가만이 관찰되었다. 최소배지에서 배양한 후 분석한 결과, $P_{180}-metX$에서의 발현양상은 배지에 첨가된 methionine에 의해 영향받지 앓음을 확인하였는데, 이는 $P_{180}$ 단편이 생합성 유전자의 derepression에 의한 아미노산 생산균의 개량에 효율적으로 이용될 수 있음을 의미한다. $P_{180}-metA$를 라이신 생산균에 도입하는 경우 최대 약 0.8g/l의 메치오닌이 생산됨을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제11권5호
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• pp.819-824
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• 2001

The principal DNA modification systems of the amino-acid-producing bacteria Corynebacterium glutamicum AS019, Brevibacterium flavum BF4, and B. lactofermentum BL1 was investigated using two approaches; digestion of plasmid DNA isolated from these species TseI and Fnu4HI, and sequence analysis of the putative methyltransferase target sites following the derivatization of DNA using metabisulfite treatment. The C. glutamicum and B. flavum strains showed similar digestion patterns to the two enzymes, indicating that the target for cytidine methyltransferase recognizes 5'-GCSGC-3'(where S is either G or C). Mapping the methylated cytidine sites by bisulfite derivatization, followed by PCR amplification and sequencing, was only possible when the protocol included an additional step eliminating any underivatized DNA after PCR amplification, thereby indicating that the derivatization was not $100\%$ efficient. This may have been due to the high G0C content of this genus. It was confirmed that C. glutamicum AS019 and B. flavum BF4 methylated the cytidine in the $Gm^5CCGC$ sequences, yet there were no similar patterns of methylation in B. lactofermentum, which was consistent with the distinctive degradation pattern seen for the above enzymes. These findings demonstrate the successful application of a modified bisulfite derivatization method with the Corynebacterium species for determining methylation patterns, and showed that different species in the geneus contain distinctive restriction and modification systems.

• 한국미생물·생명공학회지
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• 제18권3호
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• pp.296-300
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• 1990

자외선 조사와 NTG를 처리하여 Brevibacterium flavum 10AHR(arg his $Rif^r$)과 Corynebacterium glutamicum 11TS(trp $Sm^r$의 돌연변이주를 분리하였다. B.flavum 10AHR과 C.glutamicum 11TS를 300$\mu g$/ml의 lysozyme으로 18시간 처리하여 원형질체를 형성하고, 융합시 30의 PEG 6,000으로 처리하였을 때 가장 높은$3.7\times 10^{-6}$의 융합빈도를 나타내었다.