• Journal of Microbiology and Biotechnology
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• v.23 no.11
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• pp.1500-1508
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• 2013

Ebosin, a novel exopolysaccharide produced by Streptomyces sp. 139, has obvious antirheumatic arthritis activity in vivo, and its biosynthesis gene cluster (ste), consisting of 27 open reading frames, has been identified. This paper reports our study of the gene functionality of ste8, the predicted protein product of which is homologous to some bacterial chain length determinant Wzz proteins. For characterization of Ste8, ste8 was cloned and expressed in the mutant strain E. coli 086:H2 (${\Delta}wzz$). The functional complementation of wzz by ste8 was demonstrated by the restoration of wild-type lipopolysaccharide biosynthesis and increased levels of serum resistance of E. coli 086:H2 (${\Delta}wzz$) (pET30a-ste8). To examine the function of ste8 in ebosin biosynthesis, the gene was knocked out with a double crossover via homologous recombination. The molecular weight of the ebosin derivative EPS-8m produced by the mutant Streptomyces sp. 139 ($ste8^-$) was much lower than that of ebosin, and the binding activity of EPS-8m for IL-1R decreased significantly compared with ebosin. These results demonstrate that ste8 encodes a chain length determinant (Wzz) that functions in ebosin biosynthesis.

• Journal of the Korea Organic Resources Recycling Association
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• v.12 no.4
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• pp.139-147
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• 2004

We have isolated two bacterial strains, FM-02 and AL-02, which produced EPS from forest soil for the restoration of forest fire by promoting soil aggregation. FM-02 was found to be Gram negative rod and belong to Beta Proteobacterium sp. through 16s-rDNA sequence analysis, and AL-02 was Gram positive rod and showed 81% of similarity to Zoogloea sp. through the analysis of 16s-rDNA sequence. FM-02 and AL-02 produced about 1.8g and 8.3g of EPS, respectively, per 1L of culture as dry weight. Flocculation activity (FA) was also measured in two strains. FM-02 showed 2.31 FA against active carbon, and AL-02 showed 6.21 FA against kaolin clay. From these results, we expect that AL-02 strain will be applied as a good biological material for the reduction of forest soil erosion by wild and rain after fire through promoting coagulation of soil particles.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.2
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• pp.254-260
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• 2012

This study was conduced to ferment high quality wine by using Omija fruit. Dry Omija farmed and dried in the Moonkyung area was used in this study. The Omija was soaked in 10~40 folds of distilled water to extract water-soluble components and the fluid was filtered after soaking for 6 hours at $50^{\circ}C$. Strains of alcoholic yeast were isolated respectively from spoiled Omija extract. Isolated alcoholic yeasts, OM-1 and OM-2, showed a round to ellipsoidal shape and formed white or milky white colonies on a solid YM medium. Two yeasts produced 10.33~11.23% alcohol from Omija extract adjusted to $10^{\circ}Brix$ with sugar. Their abilities to ferment alcohol were higher than those of other yeast strains belonging to Saccharomyces cerevisiae such as KCTC 7296 (standard strain of Korean Biological Resources Center), Makgeolli yeast, or beer yeast. The isolates OM-1 and OM-2 showed similar abilities in alcohol fermentation. However, the wine fermented by OM-2 got a better sensory score especially with color. Growth of OM-2 was significantly accelerated by addition of a 0.1% urea and 0.02% mineral mixture. A vitamin mixture was effective for the growth only when urea was added as well.

• Journal of Life Science
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• v.18 no.11
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• pp.1507-1512
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• 2008

In spite of long research period for Salmonella typhi, little information is known about the pathogenesis mechanism of human typhoid fever caused by S. typhi due to lack of infection model in animals. A wild-type of S. typhi Ty2 strain requires cysteine to grow on minimal media. We hypothesized that this cysteine requirement may restrict colonization of S. typhi in animals during infection process. Among the S. typhi strains carrying Salmonella typhimurium genomic library, we have isolated three S. typhi transformants growing on minimal media without cysteine. Although there were three ORFs in DNA of pBP71, the STM1490 ORF complemented cysteine auxotrophy of S. typhi. Analysis of the deduced amino acid sequence of the STM1490 homolog in S. typhi revealed that there are differences in two amino acids. Plasmids containing amino acid substitutions in STM1490 supported S. typhi growth on minimal media without cysteine, indicating irrelevance of these two amino acids to STM1490 function. These results tells us that there are other factors or systems involved in cysteine requirement of S. typhi.

• Development and Reproduction
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• v.5 no.2
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• pp.151-158
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• 2001

Genomic DNA from the blood of crucian carp(Carassius carassiu) from lake and aquaculture facility in Kunsan, Korea was extracted in order to identify genetic differences by polymerase chain reaction-randomly amplified polymorphic DNAs(PCR-RAPD). Out of 12 primers, 6 generated 266 highly reproducible RAPD markers, producing approximately 2.1 polymorphic bands per primer in crucian carp from lake. The degree of similarity varied from 0.18 to 0.76 as calculated by bandsharing analysis in crucian carp from lake. The RAPD outlines obtained with DNA of two different crucian carp populations from Kunsan were different(0.47 from lake and 0.70 from aquaculture facility, respectively). The electrophoretic analysis of polymerase chain reaction-randomly amplified polymorphic DNAs(PCR-RAPD) products showed high levels of similarity between different individuals in crucian carp from aquaculture facility. This result implies the genetic similarity due to raising in the same environmental condition or inbreeding within the crucian carp from aquaculture facility in Kunsan. In other words, crucian carp may have high levels of genome DNA diversity due to the introduction of the wild population from the other sites of Knsan even if it may be the geographical diverse distribution of this species. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of important aquacultural fish species, crucian carp. However, in future, additional populations and sampling sites will be necessary to complement weak points.

• Parasites, Hosts and Diseases
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• v.29 no.2
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• pp.181-188
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• 1991

Epidemiological studies on host rodents of tsutsugamushi disease were carried out during the period of July∼September 1990 at nine localities of central Korea. Among total 111 wild rodents trapped by the modified Sherman live traps, 103 were Apodemus agrarius (92.8%), seven were Crocidura lasiura (6.3%) and one was Microtus fortis (0.9%) , showing 24.0% of trapping rate in winter, 11.7% in spring, 11,2% in summer and 12.0% in autumn. Out of 103 A. agrarius 84 were parasitized by chiggers, showing 81.6% of the infestation rate and 43.0 of the chigger index. The antibody positive rate of A. agrarius sera to Rickettsia tsutsugamushi was significantly variable by locality, being in the range of 0∼78.6%. The seasonal change of the antibody positive rate at Dorai 5-ri, Goyang-gun was 75.8% in average during November∼March, decreased to 30.3% in April and further decreased to 13.3% in average during May∼August. Among 33 antibody positives, 31 were Karp strain and two were Gilliam. Seven Crocidura lasiura sera showed all negative. R. tsutsugamushi organisms were isolated from three A. ngrarius out of 94 mice tested, showing 3.2% of the infection rate.

• Korean Journal of Microbiology
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• v.48 no.1
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• pp.1-7
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• 2012

This study explores the interaction between the production of indole-3-acetic acid (IAA), a typical phytohormone auxin and the role of IAA biosynthetic pathways in each IAA producing rhizobacterial strain. The bacterial strains were isolated from rhizosphere of wild plants and identified as Acinetobacter guillouiae SW5, Bacillus thuringiensis SW17, Rhodococcus equi SW9, and Lysinibacillus fusiformis SW13. A. guillouiae SW5 exhibited the highest production of IAA using tryptophan-dependent pathways among the 4 strains. When indole-3-acetamide (IAM) was added, Rhodococcus equi SW9 showed the highest IAA production of $3824{\mu}g/mg$ protein using amidase activity. A. guillouiae SW5 also showed the highest production of IAA using two pathways with indole-3-acetonitrile (IAN), and its nitrile hydratase activity might be higher than nitrilase. B. thuringiensis SW17 showed the lowest IAA production, and most of IAA might be produced by the amidase activity, although the nitrilase activity was the highest among 4 strains. The roles of nitrile converting enzymes were relatively similar in IAA synthesis by Lysinibacillus fusiformis SW13. Tryptophan-independent pathway of IAA production was utilized by only A. guillouiae SW5.

• Korean Chemical Engineering Research
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• v.49 no.1
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• pp.101-104
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• 2011

Cellobiose dehydrogenase(CDH) as a hemoflavoenzyme is secreted out of cell in the cellulose degradation. As CDH strongly bound to amorphous cellulose, it helps cellulose hydrolysis by cellulase. CDH may have an important role of saccharification process for bioethanol production. In this study, Phanerochaete chrysosporium ATCC 32629 was selected for the production of CDH among other strains tested. The optimal temperature and pH of CDH produced by P. chrysosporium ATCC 32629 were ${55^{\circ}C}$ and 4, respectively. To improve the activity of CDH, the mutation of P. chrysosporium was performed using proton beam that has high energy level partially. As a result, P. chrysosporium mutant with the high activity was selected at 1.2 kGy in a range of 99.9% lethal rate. The CDH and $\beta$-glucosidase activities of mutant were 1.4 fold and 20 fold higher than those of wild strain. Therefore, P. chrysosporium mutant with the high activities of CDH and $\beta$-glucosidase was obtained from mutation by proton beam irradiation.

• Microbiology and Biotechnology Letters
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• v.43 no.1
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• pp.56-64
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• 2015

It has been known for some time to the wine industry that non-Saccharomyces yeasts play an important role in increasing volatile components through the secretion of extracellular enzymes. The objective of this study was to investigate what types of enzymes are produced by 1,007 non-Saccharomyces yeast strains isolated from Korean fermented foods. Among 1,007 yeast strains, the 566, 45 and 401 strains displayed β-glucosidase, glucanase and protease activity, respectively. In addition, the 563 and 610 strains possessed tolerances against cerulenin and TFL, and the 307 strain was tolerant to 15% ethanol. Yeasts producing harmful biogenic amines and hydrogen sulfide were excluded from further study, and eventually 12 yeast strains belonging to the genera Wickerhamomyces, Hanseniaspora, Pichia, Saccharomyces were identified, based on the 26S rRNA gene sequences. Among the 12 strains, the 9 and 5 strains possessed glucose and ethanol tolerance, respectively. Yeasts belonging to the genus Saccharomyces produced more than 8% alcohol, but non-Saccharomyces yeasts produced only 3% alcohol.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.555-562
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• 2012

PCR was performed to analyze the ${\beta}$-lactamase genes carried by ampicillin-resistant Vibrio spp. strains isolated from marine environments in Korea between 2006 and 2009. All 36 strains tested showed negative results in PCR with the primers designed from the nucleotide sequences of various known ${\beta}$-lactamase genes. This prompted us to screen new ${\beta}$-lactamase genes. A novel ${\beta}$-lactamase gene was cloned from Vibrio alginolyticus KV3 isolated from the aquaculture water of Geoje Island of Korea. The determined nucleotide sequence (VAK-3 ${\beta}$-lactamase) revealed an open reading frame (ORF) of 852 bp, encoding a protein of 283 amino acids (aa), which displayed low homology to any other ${\beta}$-lactamase genes reported in public databases. The deduced 283 aa sequence of VAK-3, consisting of a 19 aa signal peptide and a 264 aa mature protein, contained highly conserved peptide segments specific to class A ${\beta}$-lactamases including the specific amino acid residues STFK (62-65), SDN (122-124), E (158), and RTG (226-228). Results from PCR performed with primers specific to the VAK-3 ${\beta}$-lactamase gene identified 3 of the 36 isolated strains as V. alginolyticus, Vibrio cholerae, and Photobacterium damselae subsp. damselae, indicating the utilization of various ${\beta}$-lactamase genes including unidentified ones in ampicillin-resistant Vibrio spp. strains from the marine environment. In a mating experiment, none of the isolates transfered the VAK-3 ${\beta}$-lactamase gene to the Escherichia coli recipient. This lack of mobility, and the presence of a chromosomal acyl-CoA flanking sequence upstream of the VAK-3 ${\beta}$-lactamase gene, led to the assumption that the location of this new ${\beta}$-lactamase gene was in the chromosome, rather than the mobile plasmid. Antibiotic susceptibility of VAK-3 ${\beta}$-lactamase was indicated by elevated levels of resistance to penicillins, but not to cephalosporins in the wild type and E. coli harboring recombinant plasmid pKV-3, compared with those of the host strain alone. Phylogenetic analysis showed that VAK-3 ${\beta}$-lactamase is a new and separate member of class A ${\beta}$-lactamases.