• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.05a
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• pp.397-401
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• 2004

장조림 원료 쇠고기 우둔살과 가열 처리한 쇠고기 우둔살에서 중온성 세균, 저온성 세균, 혐기성 세균, 포자형성균 및 대장균군 등과 같은 미생물 균수의 변화를 측정하였고, B. cereus, C. botulinum, C. perfringens, L. monocytogenes 등과 같은 9종류의 주요 병원성 세균의 분리를 시도하였다. 원료육에서 중온균, 저온균, 혐기성 균 등은 대부분 높은 분포를 보였으나 가열 처리육에서는 급격히 감소하였으며, 대장균군, E. coli, Enterpbacteriacea, C. perfringens, S. aureus 등은 원료육과 가열 처리한 쇠고기 우둔살에서 모두 검출되지 않았다. 식중독균인 B. cereus, C. perfringens 및 L. monocytogenes등 3균주가 원료육에서 분리된 반면, C. botulinum, E. coli 0157:H7, Salmonella spp., Shigella spp., S. aureus 및 Y. enterocolitica는 분리되지 않았다.

• Korean Journal of Soil Science and Fertilizer
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• v.47 no.6
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• pp.432-436
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• 2014

While searching for healthier diets, people became more attentive to agricultural organic products. However, organic foods may be more susceptible to microbiological contamination because of the use of livestock manure compost and liquid manure, potential sources of pathogenic bacteria. This study was undertaken to investigate the persistence of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes in soil, liquid manure amended soil, and liquid manure. Loamy soil, liquid manure amended soil, and liquid manure were inoculated with S. enterica, E. coli O157:H7, and L. monocytogenes. Samples were incubated in consistent moisture content at $25^{\circ}C$. Samples had been periodically collected during 120 days depending on the given conditions. S. enterica and E. coli O157:H7 survived over 120 days in loamy soil and over 60 days in liquid manure amended soil, respectively. L. monocytogenes decreased faster than other pathogens in soil. S. enterica, E. coli O157:H7, and L. monocytogenes survived for up to 5 days in liquid manure. S. enterica and E. coli O157:H7 in soil decreased by 2 to $2.5log\;CFU\;g^{-1}$ for 120 days. S. enterica and E. coli O157:H7 in liquid manure amended soil decreased slowly for 21 days. However, S. enterica, E. coli O157:H7, and L. monocytogenes sharply decreased after 21 days. S. enterica, E. coli O157:H7, and L. monocytogenes in soil increased by 0.5 to $1.0log\;CFU\;g^{-1}$ for 7 days. Foodborne pathogens in soil and liquid manure amended soil gradually decreased over time.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.155-160
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• 1986

$\alpha$-Amylase gene of B. amyloliquefaciens was cloned to E. coli-yeast shuttle vector YEp-13 and expressed in E. coli. Chromosomal DNA of B. amyloliquefaciens was partially digested with Sau3Al and YEp13 plasmid was cleaved with BamH1. The hybrid plasmid, pHA28, was constructed by shotgun method and transformed to E. coli C600 and HB101. The amount of $\alpha$-amylase produced by transformants of E. coli was about 20% to 30% of that produced by B. amyloli-quefaciens. About 65% of $\alpha$-amylase produced by transformant was secreted into periplasm and the others were located in cytoplasm. $\alpha$-Amylase production was maximal when transformants were cultivated for 15hr to 20hr. As the result of agarose gel electrophoresis, pHA28 plasmid was found to be various in its size. This result suggested that pHA28 plasmid was segregated.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.470-478
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• 2012

Recent genome comparisons of E. coli B and K-12 strains have indicated that the makeup of the cell envelopes in these two strains is quite different. Therefore, we analyzed and compared the envelope proteomes of E. coli BL21(DE3) and MG1655. A total of 165 protein spots, including 62 nonredundant proteins, were unambiguously identified by two-dimensional gel electrophoresis and mass spectrometry. Of these, 43 proteins were conserved between the two strains, whereas 4 and 16 strain-specific proteins were identified only in E. coli BL21(DE3) and MG1655, respectively. Additionally, 24 proteins showed more than 2-fold differences in intensities between the B and K-12 strains. The reference envelope proteome maps showed that E. coli envelope mainly contained channel proteins and lipoproteins. Interesting proteomic observations between the two strains were as follows: (i) B produced more OmpF porin with a larger pore size than K-12, indicating an increase in the membrane permeability; (ii) B produced higher amounts of lipoproteins, which facilitates the assembly of outer membrane ${\beta}$-barrel proteins; and (iii) motility- (FliC) and chemotaxis-related proteins (CheA and CheW) were detected only in K-12, which showed that E. coli B is restricted with regard to migration under unfavorable conditions. These differences may influence the permeability and integrity of the cell envelope, showing that E. coli B may be more susceptible than K-12 to certain stress conditions. Thus, these findings suggest that E. coli K-12 and its derivatives will be more favorable strains in certain biotechnological applications, such as cell surface display or membrane engineering studies.

• 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.

• KSBB Journal
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• v.24 no.1
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• pp.96-100
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• 2009

One of the major drawbacks associated with the high-level expression of the recombinant proteins in Escherichia coli is the formation of insoluble inclusion bodies in the cytoplasm. Production of recombinant protein at reduced temperature has proven effective in improving the solubility of a number of structurally and functionally unrelated proteins, but a major limitation of using low temperatures for recombinant protein production in E. coli is the reduced rate of synthesis of the heterologous protein caused by the significant reduction of cell growth rate. Here we investigated the effect of co-expression of CspA, a cold-shock protein known to be RNA chaperone at low temperature, on the productivity of recombinant protein at various temperatures by using green fluorescence protein (GFP) as a model recombinant protein. We could observe that the co-expression of CspA enhanced the productivity of GFP at $15^{\circ}C$ by accelerating the growth of E. coli at the temperature. On the other hand, the CspA coexpression didn't affect the cell growth rate as well as the specific GFP production rate at other tested temperatures, $20^{\circ}C$, $25^{\circ}C$, and $37^{\circ}C$.

• Journal of Food Hygiene and Safety
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• v.24 no.3
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• pp.232-237
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• 2009

This study was conducted to develop a model for describing the effect of storage temperature (4, 10, 15, 20, 25, 30 and $35^{\circ}C$) on the growth of Escherichia coli O157 : H7 in ready-to-eat (RTE) lettuce treated with or without (control) alkaline electrolyzed water (AIEW). The growth curves were well fitted with the Gompertz equation, which was used to determine the specific growth rate (SGR) and lag time (LT) of E. coli O157 : H7 ($R^2$ = 0.994). Results showed that the obtained SGR and LT were dependent on the storage temperature. The growth rate increased with increasing temperature from 4 to $35^{\circ}C$. The square root models were used to evaluate the effect of storage temperature on the growth of E. coli O157 : H7 in lettuce samples treated without or with AIEW. The coefficient of determination ($R^2$), adjusted determination coefficient ($R^2_{Adj}$), and mean square error (MSE) were employed to validate the established models. It showed that $R^2$ and $R^_{Adj}$ were close to 1 (> 0.93), and MSE calculated from models of untreated and treated lettuce were 0.031 and 0.025, respectively. The results demonstrated that the overall predictions of the growth of E. coli O157: H7 agreed with the observed data.

• Journal of Life Science
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• v.14 no.5
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• pp.868-873
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• 2004

The efficient fermentative production of L-threonine fermentation was achieved by using Escherichia coli MT201, transformed a plasmid carrying pyruvate carboxylase gene. It is an attempt to supply oxaloacetate to the L-threonine biosynthetic pathway. In order to improve the L-threonine productivity of E. coli MT201, a plasmid pPYC which is an expression vector of the pyruvate carboxylase gene of Coryne-bacterium glutamicum, was introduced. When E. coli MT/pPYC was incubated with medium containing only glucose as a carbon source, both the cell growth and L-threonine production were reduced, compared to the results from fermentation of E. coli MT201. In order to circumvent this effect, we attempted the addition of a mixed carbon source, composed of glucose and sodium citrate at a ratio of 1.5:3.5. It was shown that L-threonine production and cell growth (OD660) with E. coli MT/pPYC reached up to 75.7 g/l and 48, respectively, at incubation for 75 hr under fed-batch fermentation conditions. It is assumed that overproduction of L-threonine by anaplerotic pathway leads unbalance of TCA cycle and sodium citrate might playa role to recover normal TCA cycle.

• Microbiology and Biotechnology Letters
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• v.26 no.1
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• pp.61-67
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• 1998

Recombinant human epidermal growth factor (rhEGF) was produced by E. coli BL2l (DE3) harboring a plasmid pYHB101. The production of rhEGF was 44.5 mg/L when the E. coli BL2l (pYHB101) was cultured at 27$^{\circ}C$ for 48 hr in the modified MBL medium containing 10 $\mu\textrm{g}$/L glucose with 10 $\mu\textrm{m}$ IPTG/lactose induction at 2 hr after inoculation. It was shown that lactose is able to induce the rhEGF expression of E. coli BL2l (pYHB101) with the same efficiency as IPTG. In the batch culture system, when induced with 10 $\mu\textrm{m}$ lactose, E. coli BL2l (pYHB101) produced maximum 45 mg/L of the rhEGF at 28 hr culture in the modified MBL medium containing 10 g/L glucose. In the semi-fed batch culture system, the volumetric yield was 160 mg/L when the culture was added with 0.5% (w/v) lactose and 0.25% (w/v) yeast extract in the late logarithmic phase and 94.3% of rhEGF was secreted as soluble form. However, when the culture was added with them in the early logarithmic phase, the volumetric yield was 120 mg/L and 20.9% of rhEGF was found in cytoplasmic insoluble aggregates. It was found that the addition time of lactose was important for production of soluble rhEGF from E. coli BL21 (pYHB101).

• Applied Biological Chemistry
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• v.39 no.2
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• pp.112-117
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• 1996

Arginine decarboxylase (ADC) is the first enzyme in one of the two pathways of diamine putrescine biosynthesis in plants. The genes encoding ADC have previously been cloned from Escherichia coli, oat and tomato genome. Two degenerate oligonucleotides (17-mer) corresponding to two conserved regions of ADC were used as primers in polymerase chain reaction of rice (Oryza sativa L.) genomic DNA, and an approximately 1.0 kbp fragment was obtained. This amplified PCR product showed an open reading frame which contains 1,022 bp of nucleotide sequences. This PCR product was cloned into pGEM-originated T vector and the short 500 bp PstI digested fragment was subcloned into pGEM-3zf(+/-) vectors to facilitate sequencing. The nucleotide sequence of this PCR product showed about 74% and 70% identity with the same regions of the oat and tomato ADC cDNA sequences, respectively. The predicted amino acid sequence exhibited 45% and 62% identity with oat and tomato ADC polypeptide fragments, respectively. The sequence similarities of 34%, 47% and 38% were previously reported in oat and E. coli, tomato and oat, and tomato and E. coli ADC amino acids, respectively. Therefore, similarities and identities between rice and oat or tomato are remarkably higher than those others of the previous reports. In the highly conserved regions in both the amino acid sequence and spacing regions among the sequences of these three, rice ADC open reading frame also has the exactly same regions with the striking similarity. RNA blot analysis showed that hnc is expressed as a transcript of approximately 2.5 kbP in the rice seedling leaf tissues.