• Korean Journal of Microbiology
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• v.54 no.1
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• pp.60-68
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• 2018

A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.

• Journal of Food Hygiene and Safety
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• v.21 no.4
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• pp.213-217
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• 2006

The halophilic bacterium Vibrio vulnificus is known to be a foodborne pathogen that causes septicemia in human. V. vulnificus infection is characterized by the high fatality rates and the primary attack against a person who have underlying diseases such as liver cirrhosis. However, there is no effective treatment for V. vulnificus septicemia except for classical treatments such as antibiotics. Recently, it has been known that lipoprotein (LDL) plays a major role in the protection against infection and inflammation. Consequently in this paper we analyzed the effects of LDL on V. vulnificus septicemia. We purified V. vulnificus cytolysin, a major virulent factor of V. vulnificus infection and measured inhibitory effects of mouse serum, cholesterol, and LDL on its hemolytic activity. Next experiments were performed to investigate whether LDL has a protective role against septicemia induced by V. vulnificus in mice. Intraperitoneal injection of LDL (1mg as protein) into mice 3hr before V. vulnificus $(1\times10^6\;CFU)$ injection, and V. vulnificus -induced lethality was determined. For the determination the relationship between LDL or cholesterol and prognosis, we determined serum levels of cholesterol and lipoprotein from V. vulnificus septicemia patients (n=15) who had visited the Chonbuk National University Hospital in Chonju. V. vulnificus cytolysin -induced hemolysis of mice erythrocytes was completely inhibited by serum, cholesterol, and low-density lipoprotein. V. vulnificus- induced lethality of mice injected with LDL showed only 40% compared to 100% of control. In survival groups (n=4) of V. vulnificus septicemia patients (n=15), their serum LDL and cholesterol revealed normal levels ($153.3{\pm}40.7,\;LDL;\;190.8{\pm}16.3$, Total cholesterol). However, in death groups (n=11) showed very low levels ($35.6{\pm}13.9,\;LDL;\;59.2{\pm}15.1$, Total cholesterol). Our study indicates that cholesterol and LDL are a prognosis indicator of V. vulnificus septicemia as well as an inhibitor of virulent action of V. vulnificus cytolysin. We suggested that the serum levels of cholesterol or LDL would be major index in the treatment and prevention of V. vulnificus septicemia.

• Journal of Food Hygiene and Safety
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• v.32 no.6
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• pp.542-549
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• 2017

To evaluate bacteriological and toxicological safety, the hygienic indicator bacterium and paralytic and diarrhetic shellfish toxins in the shellfish produced in the Kamakman Area from 2012 to 2016 were investigated. Fecal coliforms and E. coli of all 194 oyster samples tested did not exceed 230 MPN/100 g. The geometric mean of the fecal coliform analyzed with the oyster samples of harvesting period was 19.6 MPN/100 g, which was more stable than the non-harvesting period (26.5 MPN/100 g). For the toxicological evaluation of the Kamakman Area, 77 oyster samples and 350 mussel samples as an indicator were analyzed. Paralytic shellfish toxins were detected very low in the range of $40{\sim}46{\mu}g$/100 g in 13 mussel samples during late April and early June, but not in oyster samples. Diarrhetic shellfish toxin was detected in 2 of 180 samples, but it was found to be below the regulation value (0.16 mg OA equ./kg). Based on the bacteriological studies, it was confirmed that the shellfish produced in Kamakman area meets the standard of shellfish hygiene of the Food Sanitation Act and meets the Grade A of the shellfish production area of EU. As the results of the paralytic and diarrhetic shellfish toxin evaluation, it was confirmed that the Kamakman Area is also toxicologically safe for shellfish production.

• The Korean Journal of Food And Nutrition
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• v.16 no.1
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• pp.7-14
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• 2003

This study mainly focused on to investigate the effects of Schizandra chinensis on the growth of a bacterium, CS6 which was isolated from kimchi. CS6 was final]y identified to lactobacillus plantarum that caused acidification of kimchi. The ethanolic extract of Schizandra chinensis(EES) inhibited the growth of L. plantarum. Minimum inhibition concentration of crude EES on L. plantarum was 62.5mg/$m\ell$. In broth culture, 5$\mu\textrm{g}$/$m\ell$ of EES completely inhibited the growth of L. plantarum during fermentation. The addition of 0.4% of EES has no apparent effect on quality including the taste and color on kimchi. It was expected that EES-containing kimchi could extend the period of preservation. Analysis of organic acids in water fractions of EES was carried out by HPLC. It is apparent that antimicrobial active fractions contained the highest concentration of succinic acid, a little tartaric acid and malic acid. Among these organic acids, succinic acid showed the strong inhibitory effect against L. plantarum CS6 in vitro. Succinic acid-containing kimchi with a concentration of 0.4 and 0.5% had the inhibitory effect on growth of L. plantarum. Inhibitory effect of EES on amylase, cellulase and pectinase was also tested. In conclusion, the present experiment demonstrated that EES inhibited the growth of L. plantarum, and various enzyme activity. EES-containing kimchi was sustained the hardness, and initial acidity during fermentation. EES was considered as the possible additive of kimchi process and EES added in kimchi increase the quality, and storage period of kimchi.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.3
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• pp.386-395
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• 2016

To develop a functional fermentation food from Rubus coreanus Miquel (Bokbunja) and chlorella mixtures, optimal lactic acid fermentation conditions were established, and quality properties based on physicochemical evaluation such as chemical compositions, free sugars, organic acids, and antibacterial activities were investigated. Regarding optimal fermentation strain selection, formation of lactic acid was best in Lactobacillus plantarum among the experimental strains (10 kinds), and the optimal fermentation temperature was $37^{\circ}C$. In addition, overall acceptability in the sensory evaluation was highest in the 5% chlorella mixture sample. Therefore, quality properties of the prepared sample under the established optimal fermentation conditions were investigated. Moisture, total sugar (dry basis), crude fiber (dry basis), and pH of fermented Rubus coreanus Miquel juice (RCM) with 5% chlorella mixture (RCM-C5) were reduced by 4.90%, 14.15%, and 0.32%, respectively, as compared with non-fermented RCM. Meanwhile, crude protein, crude fat, and crude ash (dry basis) of RCM-C5 were elevated by 13.75%, 0.18%, and 0.73%, respectively, as compared with RCM. The yellowness (b value) of color values was greater in RCM-C5 compared to RCM. The free sugar and organic acid contents of RCM-C5 were elevated by 0.97% and 616.30 mg%, respectively, as compared with RCM. In addition, the gram positive bacterium Staphylococcus aureus was elevated by 5.83% while gram negative bacteria Escherichia coli and Salmonella Typhimurium were elevated by 2.94% and 4.67%, respectively, as compared with RCM. In conclusion, the quality properties of RCM and chlorella lactic acid fermentation mixtures were improved compared with the general RCM product. Consequently, it is possible to apply fermented RCM as a functional fermentation food.

• Food Science and Preservation
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• v.15 no.5
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• pp.760-768
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• 2008

The present study investigated the effect of Leuconostoc kimchii GJ2 (Leu. kimchii GJ2), an exopolysaccharide-producing lactic acid bacterium isolated from kimchi, on serum and liver lipid metabolism in rats fed a high-cholesterol diet. Male Sprague-Dawley rats were divided into four groups: a normal diet group (ND), a high-cholesterol diet group (HCD), a high-cholesterol diet and 200 mg/kg Leu. kimchii GJ2-administered group (HCD-LKL), and a high-cholesterol diet and 400 mg/kg Leu. kimchii GJ2-administered group (HCD-LKH). No between-group differences were found in body weight gain, food intake, or food efficiency ratio. The serum GOT and ALP activities that were elevated by the high-cholesterol diet were significantly decreased after Leu. kimchii GJ2 administration. Serum HDL-cholesterol level was markedly increased in the Leu. kimchii GJ2-administered groups, whereas the serum total cholesterol and LDL-cholesterol levels were lower in the Leu. kimchii GJ2-administered animals. Liver levels of total cholesterol and triglyceride were also markedly lower in the Leu. kimchii GJ2-administered groups. In addition, increased activities of HR-LPL and TE-LPL in adipose tissue, caused by the high-cholesterol diet, fell to normal after administration of Leu. kimchii GJ2, in a dose-dependant manner. These results suggest that Leu. kimchii GJ2 isolated from kimchi exerts an antiatherosclerotic effect by reducing serum and liver cholesterol levels.

• Korean Journal of Soil Science and Fertilizer
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• v.36 no.3
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• pp.134-144
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• 2003

A rhizobacterium Pseudomonas cholororaphis O6 produced several secondary metabolites, such as phenazines, protease, and HCN that may be involved in inhibition of the growth of phytopathogenic fungi. In field study, P. chlororaphis O6 treatment on wheat seed suppressed root rot disease caused by Fusarium culmorum. The major organic acids of cucumber root exudates were fumaric acid, malic acid, benzoic acid, and succinic acid. Glucose and fructose were major monosaccharides in cucumber root exudates. The total amount of organic acids was ten times higher than that of the sugars. P. chlororaphis O6 grew well on cucumber root exudates. The dctA gene of P. chlororaphis O6 consisted of a 1,335 bp open reading frame with a deduced amino acid sequence of 444 residues, corresponding to a molecular size of about 47 kD and pI 8.2. The deduced dctA sequence has ten putative transmembrane domains, as expected of a membrane-embedded protein. Our results indicated that organic acids in cucumber root exudates may play an important role in providing nutrient source for root colonization of biological control bacteria, and the dctA gene of P. chlororaphis O6 may be an important bacterial trait that is involved in utilization of root exudates.

• Journal of Life Science
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• v.28 no.11
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• pp.1347-1353
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• 2018

The fermentation of non-digestible soy meal can convert polysaccharides into many compounds that have a wide variety of biological functions. Bacillus strains are capable of hydrolyzing non-digestible saccharides, such as melibiose, raffinose, and stachyose, found in soy meal components. A highly active ${\alpha}$-galactosidase (${\alpha}$-d-galactoside galactohydrolase, EC 3.2.1.22) was isolated from a bacterium in a traditional Korean fermented medicinal herb preparation. The isolate, T2-16, was identified as Bacillus coagulans based on its 16S rRNA sequence and biochemical properties, and the strain was named Bacillus coagulans NRR-1207. When incubated in 10%(w/v) skim milk, Bacillus coagulans NRR1207 caused a decrease in the pH of the culture medium, as well as an increase in titratable acidity and viable cell counts. This strain also showed higher activities of ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\alpha}$-glucosidase, naphthol-AS-BO-phosphohydrolase, and acid phosphatase when compared to other enzymes. It hydrolyzed oligomeric substrates, such as raffinose and stachyose, and liberated galactose, indicating that the Bacillus coagulans NRR1207 ${\alpha}$-galactosidase hydrolyzed the ${\alpha}$-1,6 glycoside linkage. These results suggest that the decreased stachyose and raffinose contents observed in fermented soy meal are due to this ${\alpha}$-galactosidase activity. Bacillus coagulans NRR1207 therefore has potential probiotic activity and could be utilized in feed manufacturing, as well as for hydrolyzing non-digestible soy meal components.

• Journal of Dairy Science and Biotechnology
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• v.37 no.2
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• pp.115-128
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• 2019

Lactic acid bacteria obtained from traditional Kimchi were selected on the basis of their caseinolytic activity and lactose usability and examined for availability as a starter in probiotic activity. Thirty-two strains were selected as lactic acid producing bacteria in BCP agar, and two strains (KC23 and KF26) with more than 90% resistance for both acid and bile salts were selected. The two strains were identified as L. plantarum (KC23) and L. paracasei (KF26) by API 50 CHL system and 16S rRNA sequence analysis. L. plantarum (KC23) was finally selected based on its biochemical characteristics for lactose and raffinose usability. Free tyrosine content increased rapidly in 10% skimmed milk medium, from $24.1{\mu}g/mL$ after 8 h to $43.9{\mu}g/mL$ after 16 h. Additionally, the caseinolytic clear zone of 12 mm of L. plantarum (KC23) was greater than the 9 mm zone of commercial L. acidophilus CSLA. The bacterium exhibited mesophilic growth and yielded $8.9{\times}10^8CFU/mL$ when incubated at $37^{\circ}C$ for 12 h at pH 4.25. Moreover, L. plantarum KC23 exhibited antibacterial activity as it formed a clear zone of 8-13 mm for the 5 pathogens. Adherent activity was 2.23 fold higher than that of LGG. The acidity of 10% skimmed milk fermented for 12 h was 0.74%.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.369-378
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• 2018

This study was conducted to evaluate effects of dietary microbial probiotics on the growth and disease resistance of olive flounder (Paralichthys olivaceus) in a recirculating aquaculture system (RAS), and the effects of the probiotic bioaugmentation on the microbial community structure and water quality. For the analysis, 80 juvenile fish (average weight, $25.7{\pm}7.6g$; average length, $15.2{\pm}1.7cm$) were fed a basal diet containing a commercial microbial product CES-AQ1 (CES; $1{\times}10^9\;CFU/kg$ diet) in an RAS for 8 weeks. Weight gain, the specific growth rate, feed efficiency, and protein efficiency ratio of the fish fed the CES diet in the RAS were 1.5~2.5 times higher than those of fish fed the basal diet alone, or the basal diet containing oxytetracycline (OTC), yeast plus bacterium, or Bacillus subtilis in a still water system. There was no significant difference in the pathogen challenge test between fish fed the OTC diet and fish fed the CES diet in the RAS, suggesting the CES-AQ1 probiotic used in the RAS as a potential replacement for antibiotics. The RAS biofilter maintained the highest microbial diversity and appeared to harbor microbial communities with ammonium oxidation, denitrification, and fish pathogen suppression functions. Ammonia, which is hazardous to fish, was significantly decreased to < 0.5 mg/L in 19 days, indicating the effectiveness of probiotic supplementation to maintain good water quality in RAS. These results suggest that the intestinal microbial communities of fish are stabilized by a probiotic-containing diet (CES) and that bioaugmentation with probiotics may be an eco-friendly and economical supplement for aquaculture of olive flounder, promoting both good water quality and fish health in an RAS.