• Microbiology and Biotechnology Letters
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• v.19 no.5
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• pp.433-438
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• 1991

The plasmid pSY130-14 for the high production of phenylalanine is a temperaturecontrollable expression vector composed of the $P_R$ and the $P_L$ promoter and a temperature sensitive repressor, $cI_{857}$ of bacteriophage lambda. Strain AT2471 harbouring plasmid pSY13O- 14 is induced the phenylalanine production by shifting up the incubation temperaure to $38.5^{\circ}C$. Plasmid stability of E. coli AT2471 harbouring pSY130-14 was very low, it was about 30% after 48 h cultivation at $38.5^{\circ}C$ without kanamycin. The plasmid disappeared immediately at $40^{\circ}C$ without kanamycin, and at $40^{\circ}C$ adding kanamycin, the plasmid stability decreased at the beginning, but rose with the extension of the culture time. For the improvement of plasmid stability, the plasmid obtaind was designated as pSY15O-1 by changing origin region (ori) pACYC 177 of pSY130-14 for ori pSC101. E. coli AT2471 harbouring pSY150-1 was stable at $38.5^{\circ}C$ without tetracycline, and the plasrnid stability was about 40% after 48 h cultivation at $40^{\circ}C$.

• Journal of Food Hygiene and Safety
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• v.22 no.3
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• pp.151-158
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• 2007

Among 68 strains of lactic acid bacteria (LAB) isolated from Dongchimi, a strain K11 was selected due to its bactericidal activity against Escherichia coli O157 The strain K11 was identified as Lactobacillus plantarum, based on physiological and biochemical characteristics. In the late exponential phase, La. plantarum K11 showed maximum bacteriocin activity (12,800 BU/mL) and maintained until the early stationary phase. The bacteriocin activity was completely inactivated by all the proteolytic enzymes such as pepsin, protease, proteinase K, papain, chymotrypsin, and trypsin, but the activity was not affected by catalase, a-amylase, lysozyme, and lipase, suggesting proteinaceous nature of the bacteriocin. Additionally, this activity was not affected in the pH range from 3.0 to 9.0 and under storage conditions like 30 days at -20,4, or $25^{\circ}C$. Although the bacteriocin activity was absolutely lost after 15 min treatment at 121, it was relatively stable at $70^{\circ}C$ for 60 min or $100^{\circ}C$ for 30 min. The activity was disappeared by treatment with acetone, benzene, ethanol, or methanol, but it was not affected by treatment with chloroform or hexane. The antibacterial activity of the bacteriocin was good against some LAB including Lactobacillus spp., Enterococcus spp., and Streptococcus spp., but not against food-borne pathogens such as Bacillus spp., Listeria spp., and Staphylococcus spp. as well as yeasts and molds. Especially, some intestinal bacteria such as Enterobacter aerogenes and E. coli were significantly affected by the bacteriocin of La, plantarum K11. Furthermore, the addition of 640 BU/mL resulted in the complete clearance of E. coli O157 after 10 hr.

• Journal of Ginseng Research
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• v.7 no.2
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• pp.102-107
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• 1983

The effects of saponin, one of major components (Panax ginseng C.A. Meyer), on the growth of E. coli K-12 and the formation of siderphore was observed The following results were obtained. 1. When E. coli was grown on medium containing 1${\times}$10-5%-11${\times}$10-1% of the saponin, the rate of growth was stimulated at 10-1% of the saponin significantly compared to that of control. 2. When E. coli K-12 was grown on medium containing 1${\times}$10-1% of the saponin, the amount of siderphore was two times as much as the control. 3. The growth of E. coli was observed to be dependent on the concentration of siderophore when siderophore was added to medium. 4. The effect of saponin on the formation of siderophore in vitro was observed to reach maximum at 1${\times}$10-3% of the saponin. Such results suggest that the growth rate of E. coli K-12 could be enhanced by ginseng saponin fraction through stimulation of siderphore formation. We have described the fast growth of E. coli, K-12 and B. subtilis, rapid uptake of 14C-glucose, and high level of other metabolites such as lipids and proteins of E. coli, and B. subtilis in medium containing saponing fraction compared to that of microorganisms without saponin fraction.1∼3Such differences were claimed to be due to rapid uptake of 14C-glucose by widened periplasmic region throught unknown mechanism in the prescence of saponin fraction in medium3 and have raised a question whether there is another possible factor, siderophore4(Greek for iron bears), since microorganisms must secure a sufficient amount of iron for normal growth. These are known to be synthesized by the cells under iron-deficient condition and in most case, excreted into the medium5, where they can complex and solubilize any iron present there. It is generally believed that these complexes are then taken into the cells presumably by specific transport systems, thus providing iron for cell metabolism. Within the group of enteric bacteria, only three species (E. coli, S. typhimurium, and A. aerogense) have, so far, been studied in a ny detail. The main iron-binding compound produced by these species is enterochelin, and its role in iron transport is now well established. And biosynthesis of enterochelin from 2, 3- dihydroxybenzoate and serine in the prescence of magnesium ions and ATP was reported6. 2, 3-dihydroxybenzoate was also shown to involve isochorismate and 2, 3-dihydro-2, 3-dihydroxybenzoate as intermediate.7∼11 The present paper deals with the effect of ginseng saponin fraction on growth, the level of enterochelin formation in vivo and the conversion of 2, 3-dihydroxybenzoate and serine into entrochelin in vitro, and entrochelin obtained on the growth in relation to possible explanation of ginseng saponin fraction on the rapid growth of E. coli, K-12.

• Food Science and Biotechnology
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• v.17 no.4
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• pp.878-882
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• 2008

The efficacy of citric acid-aqueous chlorine dioxide ($ClO_2$) treatment of radish seeds artificially contaminated with Escherichia coli was studied. Radish seeds were inoculated with E. coli. Following inoculation, samples were stored at $4^{\circ}C$ and soaked in citric acid or aqueous $ClO_2$ for 10 min. The treatment of radish seeds using 200 ppm aqueous $ClO_2$ solution caused a 1.5 log CFU/g reduction in the population of E. coli. Compared to the aqueous $ClO_2$ treatment, soaking radish seeds in 2.0% citric acid solution for 10 min was more effective in reducing E. coli populations on radish seeds. The efficacy of spray application of chlorine (100 ppm) or 0.5% citric acid to eliminate E. coli during the germination and growth of radish was investigated. Radish seed inoculated with E. coli was treated for the duration of the growth period. Although it resulted in a decrease in the E. coli population, the spray application of 100 ppm chlorine during the growth period was not significantly effective. In contrast, the combined treatment of seeds using 200 ppm aqueous $ClO_2$ and treatment of sprouts with 0.5% citric acid solution during sprout growth was hardly effective in eliminating E. coli.

• Microbiology and Biotechnology Letters
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• v.31 no.2
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• pp.111-116
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• 2003

The xylA gene of Bacillus stearothermophilus No. 236 encoding $\beta$-xylosidase was cloned and sequenced previously. The transcriptional start site of the xylA gene cloned in E. coli was identified to be the guanine (G) by primer extension analysis. This supports that the expression of xylA gene is also directed in the E. coli cells by the previously determined transcription initiation signals, -10 sequence (CATAAT) and -35 sequence (TTGTTA) separated by 12 bp. To increase the expression of $\beta$-xylosidase, firstly the spacer region of xylA promoter was extended from 12 to 17 bp, and then the -10 and -35 elements were converted into their respective consensus sequences. The mutant promoters thus obtained were tested for their activities in both the E. coli and B. subtilis host cells. The change of the length of the spacer region from 12 to 17 bp resulted in a 1.6- and 2.5-fold increase in promoter strength in comparison with the wild type promoter in E. coli and B. subtilis cells, respectively. Also, strength of the promoter with the fourth T to A transversion on its -35 element increased in the transcription level by about 35 times compared with that of wild-type promoter. However, surprisingly the 5' end C-to-T transition of the -10 hexamer showed a 5- to 15-fold reduction in $\beta$-xylosidase activity in both E. coli and B. subtilis. Together, the present data demonstrated that the 5' end nucleotide C of the -10 sequence CATAAT and the fourth nucleotide A of the -35 hexamer are two most critical nucleotides for the promoter activity in the context of the xylA promoter.

• Journal of Food Hygiene and Safety
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• v.28 no.2
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• pp.168-173
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• 2013

This study was carried out to develop and validate predictive models of E. coli O157:H7 growth. Growth data of E. coli O157:H7 in Paprika were collected at 12, 24, 30 and $36^{\circ}C$. The population increased into 3.0 to 3.8 log10 CFU/g within 4 days, then continued to increase at a slower rate through 10 days of storage at $12^{\circ}C$. The lag time (LT) and maximum specific growth rate (SGR) obtained from each primary model was then modeled as a function of temperature using Davey and square root equations, respectively. For interpolation of performance evaluation, growth data for a mixture of E. coli O157:H7 were collected at time intervals in paprika incubated at the different temperatures, which was not used in model development. Results of model performance for interpolation data demonstrated that induced secondary models showed acceptable goodness of fit. Relative errors in the LT and SGR model for interpolation data (18 and $27^{\circ}C$) was 100%, which show acceptable goodness of fit and validated for interpolation. The primary and secondary models developed in this study can be used to establish tertiary models to quantify the effects of temperature on the growth of E. coli O157:H7 in paprika.

• Food Science and Preservation
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• v.16 no.1
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• pp.128-133
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• 2009

We performed an experiment of keeping the extension of raw and semi-dried flounder (Pleuronectes herzensteini). The effect of with (WG) or without gill (OG), drying degree (20% drying: 20D, 40% drying: 40D) and storage temperature($5^{\circ}C$) and 0.1% chitosan-ascorbate (CA) treatment of vacuum packaging flounder on the growth of contaminated microorganism during storage for 10 days were investigated. Total aerobacter (TA) in the OG-treated raw flounder was $0.29{\sim}0.44$ log cycle lower than that of WG-treated flounder. Also, the number of Staphylococcus aureus (SA) and E. coli (EO) in OG were lower compared with WG. The number of TA, SA and EO in 20 D among 0 D, 20 D and 40 D stored at $5^{\circ}C$ were lowest. Especially, the SA and EO was $0.13{\sim}0.53$, 0.3-0.88, and 0.13-0.74 log cycle lower compared with raw flounder. The growth of TA, SA and EO separated from raw flounder in tryptic soy broth were completely inhibited by 0.1% CA. The anti-biotical effect of CA of two microorganisms SA and EO that separated from flounder, and the growth of all of them were 90% (SA), 96% (EO) inhibited at the 0.1% CA. The inhibition times at $37^{\circ}C$ in soy broth was 36 hr. However when CA was added directly to flounder, it appeared inhibition effect to 0.88 log cycle. The effect of CA was better when gills removed and 20% drying.

• Journal of Microbiology and Biotechnology
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• v.6 no.5
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• pp.309-314
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• 1996

Alcaligenes sp. SH-69 can synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from a single carbon source such as glucose. To clone the phaC gene from Alcaligenes sp. SH-69, a polymerase chain reaction was performed using the oligomers synthesized based on the conserved regions of the phaC genes from other bacteria. A PCR product (550 bp) was partially sequenced and the deduced amino acid sequence was found to be homologous to that of the phaC gene from Alcaligenes eutrophus. Using the PCR fragment Southern blotting of Alcaligenes sp. SH-69 genomic DNA digested with several restriction enzymes was carried out. To prepare a partial genomic library, about 5-Kb genomic DNA fragments digested with EcoRI, which showed a positive signal in the Southern blotting, were eluted from an agarose gel, ligated with pUC19 cleaved with EcoRI, and transformed into Escherichia coli. The partial library was screened using the PCR fragment as a probe and a plasmid, named pPHA11, showing a strong hybridization signal was selected. Restriction mapping of the insert DNA in pPHA11 was performed. Cotransformation into E. coli of the plasmid pPHA11 and the plasmid pPHA21 which has phaA and phaB from A. eutrophus resulted in turbid E. coli colonies which are indicative of PHA accumulation. This result tells us that the Alcaligenes sp. SH-69 phaC gene in the pPHA11 is functionally active in E. coli and can synthesize PHA in the presence of the A. eutrophus phaA and phaB genes.

• Preventive Nutrition and Food Science
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• v.5 no.3
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• pp.131-135
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• 2000

The inhibitory effect of various organic acids on the growth and survival of Escherichia coli O157:H7 in tryptic soy broth with 0.6% yeast extract at 37$^{\circ}C$ or 4$^{\circ}C$ was determined. Minimal inhibitory pHs of acetic acid, citric acid, fumaric acid, hydrochloric acid and lactic acid were 5.0, 4.0, 4.5, 4.0 and 4.5, respectively. Acetic acid (0.012 m) showed the strongest antimicrobial activity, based on the pH values or equivalent molar concentrations, followed by lactic acid (0.0006 M), fumaric acid (0.004M) and citirc acid (0.004 M), respectively, E. coli O157:H7 with an initial inoculum of {TEX}$10^{7}${/TEX} CFU/ml and {TEX}$10^{5}{/TEX} CFU/ml in tryptic soy broth supplemented with 0.6% yeast extract, acidified to target pH with citric, fumaric and lactic acids at 37$^{\circ}C$, was completely inactivated after 7 d and 5 d incubation, respectively, except for the acetic acid (9 d). The bactericidal effect decreased at the same pH when the incubation temperature a was reduced from 37$^{\circ}C$ to 4$^{\circ}C$. The pH values of 0.2% acetic (pH 5.1), 0.6% citric (pH 4.2) and 0.4% lactic acid (pH 4.3) in TSBYE were almost correspondent to the minimal inhibitory pH values on E. coli O157:H7 of acetic (pH 4.0), citric (pH 4.0) and lactic acids (pH 4.5).

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.933-938
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• 2006

Decaprenyl diphosphate synthase (DPS) is the key enzyme for the production of coenzyme $Q_{10}$ ($CoQ_{10}$). A dps gene from Sinorhizobium meliioti KCCM 11232 (IFO 14782) was isolated by PCR and then cloned in Escherichia coli. DNA sequencing analysis revealed an open reading frame of 1,017 bp encoding a 338-amino-acid protein. The protein was identical at the 98% level to the putative octaprenyl diphosphate synthase (IspB) of S. meliloti 1021. The deduced amino acid sequence included the DDxxD domains conserved in the majority of the prenyl diphosphate synthases. Heterologous expression in E. coli BL21 (DE3) was carried out, and the $CoQ_{10}$ produced was then analyzed by HPLC. E. coli BL21 (DE3) harboring the dps gene from S. melioti produced CoQ$_{10}$ in addition to endogenous coenzyme Q$_8$ (CoQ$_8$), whereas wild-type E. coli BL21 (DE3) host did not have the ability of producing CoQ$_{10}$. The results suggest that the putative dps from S. meliloti KCTC 2353 encoded the DPS.