• Korean Journal of Microbiology
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• v.48 no.2
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• pp.109-115
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• 2012

Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.

• Journal of Food Hygiene and Safety
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• v.27 no.1
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• pp.103-107
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• 2012

We compared between an automated most-probable-number technique $TEMPO^{(R)}$TVC and traditional plating methods $Petrifilm^{TM}$ for estimating populations of total aerobic bacteria in various livestock products. 257 samples randomly selected in local retail stores and 87 samples inoculated with $E.$ $coli$ ATCC 25922, $Staphylococcus$ $aureus$ ATCC 12868 were tested in this study. The degree of agreement was estimated according to the CCFRA (Campden and Chorleywood Food Research Association Group) Guideline 29 and the agreement indicates the difference of two kinds methods is lower than 1 log base 10($log_{10}$). The samples of hams, jerky products, ground meat products, milks, ice creams, infant formulas, and egg heat formed products were showed above 95% in the agreement of methods. In contrast, proportion of agreement on meat extract products, cheeses and sausages were 93.1%, 92.1%, 89.1%, respectively. One press ham and five sausages containing spice and seasoning, two pork cutlets containing spice and bread crumbs, two meat extract product and two natural cheeses and one processing cheese with a high fat content, and one ice cream containing chocolate of all samples showed the discrepancy. Our result suggest that $TEMPO^{(R)}$TVC system is efficient to analyses total aerobic bacteria to compare manual method in time-consuming and laborious process except livestock products having limit of detection.

• Journal of Life Science
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• v.21 no.1
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• pp.119-126
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• 2011

There is a growing recognition of the significance of $H_2S$ as a biological signaling molecule involved in vascular and nervous system functions. In mammals, two enzymes in the transsulfuration pathway, cystathionine ${\beta}$-synthase (CBS) and cystathionine ${\gamma}$-lyase (CGL), are believed to be chiefly responsible for $H_2S$ biogenesis. Genetic inborn error of CGL leads to human genetic disease, cystathioninuria, by accumulating cystathionine in the body. This disease is secondarily associated with a wide range of diseases including diabetes insipidus and Down's syndrome. Although the human CGL (hCGL) overexpression is essential for the investigation of its function, structure, reaction specificity, substrate specificity, and protein-protein interactions, there is no clear report concerning optimum overexpression conditions. In this study, we report a detailed analysis of the overexpression conditions of the hCGL using a bacterial system. Maximum overexpression was obtained in conditions of low culture temperature after inducer addition, performing low aeration during overexpression, and using a low concentration inducer (0.1 mM, IPTG) for induction. Expressed hCGL was purified by His-tag affinity column chromatography and confirmed by Western blot using hCGL antibody and enzyme activity analysis. We also report that the His tag with TEV site attached protein exhibits 76% activity for ${\alpha}-{\gamma}$ elimination reaction with L-cystathionine and 88% for ${\alpha}-{\beta}$ elimination reaction with L-cysteine compared to those of wild type hCGL, respectively. His tag with TEV site attached protein also exhibits a 420 nm absorption maximum, which is attributed to the binding cofactor, pyridoxal 5'-phosphate (PLP).

• Journal of Food Hygiene and Safety
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• v.29 no.2
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• pp.79-84
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• 2014

This study has been performed for about 270 days at analyzing biologically hazardous factors in order to develop HACCP system for the non heat-frozen carrot juice. A process chart was prepared by manufacturing process of raw agricultural products of non heat-frozen carrot juice, which was contained water and packing material, storage, washing, cutting, extraction of the juice, internal packing, metal detection, external packing, storage and consignment (delivery). As a result of measuring Coliform group, Staphylococcus aureus, Salmonella spp., Bacillus cereus, Listeria Monocytogenes, Enterohemorrhagic E. coli before and after washing raw carrot, Standard plate count was $4.7{\times}10^4CFU/g$ before washing but it was $1.2{\times}10^2CFU/g$ detected after washing. As a result of testing airborne bacteria (Standard plate count, Coliform group, Yeast and Fungal) depending on each workplace, number of microorganism of in packaging room, shower room and juice extraction room was detected to be 10 CFU/Plate, 60 CFU/Plate, 20 CFU/Plate, respectively. As a result of testing palm condition of workers, as number of Standard plate count, Coliform group and Staphylococcus aureus was represented to be high as $6{\times}10^4CFU/cm^2$, $0CFU/cm^2$ and $0CFU/cm^2$, respectively, an education and training for individual sanitation control was considered to be required. As a result of inspecting surface pollution level of manufacturing facility and devices, Coliform group was not detected in all the specimen but Standard plate count was most dominantly detected in scouring kier, scouring kier tray, cooling tank, grinding extractor, storage tank and packaging machine-nozzle as $8.00{\times}10CFU/cm^2$, $3.0{\times}10CFU/cm^2$, $4.3{\times}10^2CFU/cm^2$, $7.5{\times}10^2CFU/cm^2$, $6.0{\times}10CFU/cm^2$, $8.5{\times}10^2CFU/cm^2$ respectively. As a result of analyzing above hazardous factors, processing process of ultraviolet ray sterilizing where pathogenic bacteria may be prevented, reduced or removed is required to be controlled by CCP-B (Biological) and critical level (critical control point) was set at flow speed is 4L/min. Therefore, it is considered that thorough HACCP control plan including control criteria (point) of seasoning fluid processing process, countermeasures in case of its deviation, its verification method, education/training and record control would be required.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.21-26
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• 2010

As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.

• Journal of Korean Society of Environmental Engineers
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• v.39 no.5
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• pp.237-245
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• 2017

This study was aimed at investigating effluent water quality and proposing reuse possibilities for 12 sewage treatment plants discharged more than $5,000m^3/day$ in order to recycle the sewage treatment plant effluent of Jeollabuk-do effectively. Additionally, a laboratory scale test for reprocessing water discharge was performed. Categories of reclaimed sewage water reuse were divided into 7 topics and analyzed a total of 28 items including 16 heavy metals based on water quality standard. As a results, color, BOD, TN, chloride and Electrical Conductivity (EC) exceeded reused water quality standard. In particular, color and TN exceeded in 8 and 5 sewage treatment plants, respectively. The value of chloride and EC were high in sewage treatment plants including the food and industrial wastewater. At 4 sewage treatment plants were possible to reuse without re-treatment. The laboratory scale test was conducted to SandFilter (SF)-Granular Activated Carbon (GAC)-MicroFiltraion (MF)-Reverse Osmosis (RO). Both the removal efficiencies and economic feasibility of total E. coli., color and Suspended Solid (SS) suited in case using the SF-GAC treatment method. The removal of chloride and EC had little effect in the case of SF-GAC-MF system, but RO showed over 90% of removal efficiency. After using SF-GAC process only, the concentration of $UV_{254}$ decreased sharply from 0.3651 /cm to 0.0306 /cm and it showed over 92% of removal efficiency rate. In conclusion, for the effective reuse of sewage discharged water, water quality and the surrounding terrain of treatment plants should be all taken into account. If it needed for the reprocessing, both the selection for treatment and economic combination treatment methods will have to be considered.

• Journal of Navigation and Port Research
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• v.28 no.10 s.96
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• pp.899-908
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• 2004

The rise of income and introduction of 5 day a week working system give korean people opportunities to enjoy their leisure time. And many korean people have much interest in oceanic sports such as yachting and also oceanic leisure equipments. With the popularization and development of the equipments, the scope of oceanic activities has been expanding in Korea just as in the advanced oceanic countries. However, The current conditions for the sports in Korea are not advanced and even worse than underdeveloped countries. In order to develop the underdeveloped resources of Korean marina, we need to customize the marina models of advanced nations to serve the specific needs and circumstances of Korea As such we have carried out a comparative analysis of how Austrailia, Newzealand, Singapore, japan and Malaysia operate their marina, reaching the following conclusions. Firstly, in marina operations, in order to protect personal property rights and to preserve the environment, we must operate membership and non-membership, profit and non-profit schemes separately, yet without regulating the dress code entering or leaving the club house. Secondly, in order to accumulate greater value added, new sporting events should be hosted each year. There is also the need for an active use of volunteers, the generation of greater interest in yacht tourism, and the simplification of CIQ procedures for foreign yachts as well as the provision of language services. Thirdly, a permanent yacht school should be established, and classes should be taught by qualified instructors. Beginners, intermediary, and advanced learner classes should be managed separately with special emphasis on the dinghy yacht program for children. Fourthly, arrival and departure at the moorings must be regulated autonomically, and there must be systematic measures for the marina to be able, in part, to compensate for loss and damages to equipment, security and surveillance after usage fees have been paid for. Fifthly, marine safety personnel must be formed in accordance with Korea's current circumstances from civilian organizations in order to be used actively in benchmarking, rescue operations, and oceanic searches at times of disaster at sea.

• Journal of Food Hygiene and Safety
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• v.25 no.4
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• pp.367-375
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• 2010

The bacteriological and physiochemical analysis of sea water and sediments in Tongyeong harbor was conducted to evaluate sanitary conditions. The samples were collected at 8 stations established once a month from June, 2008 to May, 2009. During the study period, the range of temperature was from 6.7 to $25.2^{\circ}C$, transparency ranged from 1.2 to 2.6 m, chemical oxygen demand ranged from 1.90 to 2.92 mg/L, dissolved oxygen ranged from 6.2 to 10.5 mg/L, dissolved nitrogen ranged from 0.052 to 0.098 mg/L, phosphate ranged from 0.044 to 0.065 mg/L, respectively. Seafood, if eaten raw, carries the risk of food poisoning. Seafood poisoning is often cause by pathogenic microorganism originating from fecal contamination, such as Salmonella sp., Shigella sp. and norovirus. Fecal coliforms are an important indicator of fecal contamination. Therefore, data on fecal coliform are very important for evaluating the safety of fisheries in coastal areas. So, we investigated the sanitary indicate bacteria. The coliform group and fecal coliform MPN's of sea water in Tongyeong harbor were ranged from < 1.8~22,000/100 mL (GM 164.9 MPN/100 mL) and < 1.8~7,900 MPN/100 mL (GM 33.7 MPN/100 mL), respectively. Total coliform were detected 97.0% in 96 of samples and 68.9% of total coliforms were fecal coliforms. These results similar to another seawater detection ratio of total coloforms and fecal coliforms. The Vibrios was isolated and identified with VITEK system. Four hundred eighty strains that were obtained from sea water samples in Tongyeong harbor Detection ratio Vibrio alginolyticus, 34.2%, Vibrio parahaemolyticus, 13.8%, Vibrio vulnificus 10.0%, and V. mimicus 12.5% respectively. Vibrio cholerae O1, was not detected. During the study period, the ranges of water content, ignition loss, COD, and acid volatile sulfates in sediments in Tongyeoung harbor were 41.0~57.4%, 7.8~10.5%, 6.51~9.30 mg/g, 0.04~0.09 mg/g, respectively. Heavy metals in sediment of Tongyeoung harbor were Cd, $0.10{\pm}0.05$; Cu, $4.79{\pm}8.20$; As, $1.95{\pm}0.17$; Hg, $0.10{\pm}0.07$; $Cr^{6+}$, $0.34{\pm}0.12$; Zn, $125.33{\pm}16.40$; Ni, $16.43{\pm}1.93$ mg/kg.

• Journal of Life Science
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• v.31 no.3
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• pp.356-365
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• 2021

Recently, we sequenced the entire genome of a freshwater agar-degrading bacterium Cellvibrio sp. KY-GH-1 (KCTC13629BP) to explore genetic information encoding agarases that hydrolyze agarose into monomers 3,6-anhydro-L-galactose (L-AHG) and D-galactose. The KY-GH-1 strain appeared to possess nine β-agarase genes and two α-neoagarobiose hydrolase (α-NABH) genes in a 77-kb agarase gene cluster. Based on these genetic information, the KY-GH-1 strain-caused agarose degradation into L-AHG and D-galactose was predicted to be initiated by both endolytic GH16 and GH86 β-agarases to generate NAOS (NA4/NA6/NA8), and further processed by exolytic GH50 β-agarases to generate NA2, and then terminated by GH117 α-NABHs which degrade NA2 into L-AHG and D-galactose. More recently, by employing E. coli expression system with pET-30a vector we obtained three recombinant His-tagged GH50 family β-agarases (GH50A, GH50B, and GH50C) derived from Cellvibrio sp. KY-GH-1 to compare their enzymatic properties. GH50A β-agarase turned out to have the highest exolytic β-agarase activity among the three GH50 isozymes, catalyzing efficient NA2 production from the substrate (agarose, NAOS or AOS). Additionally, we determined that GH117A α-NABH, but not GH117B α-NABH, could potently degrade NA2 into L-AHG and D-galactose. Sequentially, we examined the enzymatic characteristics of GH50A β-agarase and GH117A α-NABH, and assessed their efficiency for NA2 production from agarose and for production of L-AHG and D-galactose from NA2, respectively. In this review, we describe the benefits of recombinant GH50A β-agarase and GH117A α-NABH originated from Cellvibrio sp. KY-GH-1, which may be useful for the enzymatic hydrolysis of agarose for mass production of L-AHG and D-galactose.