• Korean Journal Plant Pathology
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• v.2 no.2
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• pp.121-128
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• 1986

The biochemical properties of ribonucleic acid (RNA) and coat protein of the mild tobacco mosaic virus (TMV) mutant, Tw 333 are described. The molecular weight of the RNA calculated from polyacrylamide gel electrophoresis was $2.03\times10^6$ daltons. The molar ratio of the bases of the RNA was 25.4 guanine, 29.2 adenine, 17.5 cytosine and 27.9 uracil in moles. The hyperchromicity on Tw 333-RNA by thermal denaturation was $25.1\%$, indicating Tm value of $47^{\circ}C$. The virus coat protein migrated as a single component in SDS-polyacrylamide gel electrophoresis and had a molecular weight of 17,500 daltons. A total of 158 amino acid residues are present in the protein. Separation of the tryptic peptides by electrophoresis and chromatography yielded ninhydrin-positive compounds. The biochemical properties of RNA and coat protein of the mild mutant we very similar to those of wild type of TMV-OM strain, but some difference between the strains were observe in the base composition, hyperchromicity, amino acid composition and tryptic peptide map.

• The Journal of Korean Society of Virology
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• v.28 no.4
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• pp.337-345
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• 1998

Hantaviruses are distributed in rodent population world-widely even in geographical areas where hemorrhagic fever with renal syndrome (HFRS) has not been reported. Various species of Family Muridae and Arvicolidae serve as the natural reservoirs of hantaviruses. Hantaan virus, Seoul virus, Puumala virus, Prospect HII virus, Sin Nombre virus and New York virus are members of genus Hantavirus and isolated from lungs of A. agrarius, R. norvegicus, C. glareolus, M. pennsylvanicus, P. maniculatus and P. leucopus respectively. This experiment was intended to find the distribution of hantavirus infection among wild rodents and isolate the hantavirus from lung tissue of seropositve Apodemus peninsulae, and compared the nucleotide and amino acid sequences with prototype of hantaan virus 76-118 strain. Hantaviral sequences were amplified from lung tissues of A. peninsulae by reverse-transcriptase polymerase chain reaction. Alignment and comparison of the 324 nucleotide of G2 region of M-genomic segment diverged 4.6% and 0% at the nucleotide and amino acid levels, and complete N protein-coding region of S-genomic segment diverged 3.7% and 1.4% nucleotide and amino acid levels, respectively. This is the report to spill-over on the hantaan virus from A. agrarius to A. peninsulae in Korea.

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.560-567
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• 1998

We have found that Thermoactinomyces vulgaris KFB-Cl00 produces a chitinase. The optimum temperature and pH of the enzyme activity were $55^{\circ}C$ and 6.5. The enzyme was stable after heat treatment at $80^{\circ}C$ for 30 min and stable in acidic and basic conditions (PH 6.0~11.0). The thermostable endo-chitinase from Thermoactinomyces vulgaris KFB-C100 was cloned into the plasmid pBR322 by using E. coli DH5$\alpha$ as a host strain. The positive clone carrying a recombinant plasmid (PKCHI23) with a 4.1-kb fragment containing the chitinase gene was found. The recombinant plasmid was analyzed to determine the essential region for chitinase activity and obtained a 2.3-kb fragment, which was sub cloned into pTrc99A using the PstI and SalI sites to construct pTrc99A/pKCHI23-3. The resulting plasmid exerted high chitinase activity upon transformation of E. coli XL1-Blue cells. Chitinase was overproduced 14 times more in the clone cells than in the wild-type cells and the enzyme was purified to homogeneity. The purified enzyme showed the similar properties as the native chitinase from T. vulgaris in terms of molecular weight and substrate specificity. The catalytic action of the cloned enzyme was an endo type, producing chitobiose as a major reaction product.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1244-1249
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• 2009

Endophytic fungi are known for the production of valuable metabolites, but information on the gibberellin production capacity of this group is limited. We isolated 9 endophytic fungi from the roots of salt-stressed soybean plants and screened them on waito-c rice, in order to identify plant growth promoting fungal strains. The fungal isolate TK-2-4 gave maximum plant length (20.35 cm) promotion in comparison with wild-type Gibberella fujikuroi (19.5 cm). In a separate experiment, bioassay of TK-2-4 promoted plant length and biomass of soybean cultivar Taegwangkong. The TK-2-4 culture filtrate was analyzed for the presence of gibberellins, and it was found that all physiologically active gibberellins, especially $GA_4$ and $GA_7$, were present in higher amounts ($GA_1$, 0.11 ng/ml; $GA_3$, 2.91 ng/ml; $GA_4$, 3.21 ng/ml; and $GA_7$, 1.4 ng/ml) in conjunction with physiologically inactive $GA_9$ (0.05 ng/ml), $GA_{12}$ (0.23 ng/ ml), $GA_{15}$ (0.42 ng/ml), $GA_{19}$ (0.53 ng/ml), and $GA_{20}$ (0.06 ng/ml). The fungal isolate TK-2-4 was later identified as a new strain of Phoma herbarum, through the phylogenetic analysis of 28S rDNA sequence.

• Journal of Microbiology and Biotechnology
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• v.20 no.8
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• pp.1251-1258
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• 2010

During the nitrogen-fixing process, ammonia ($NH_3$) is incorporated into glutamate to yield glutamine and is generally not secreted. However, in this study, $NH_3$-excreting strains of nitrogen-fixing Paenibacillus were isolated from soil. The ammonium production by the Paenibacillus strains was assayed in different experiments (dry biomass, wet biomass, cell-free extract, and cell-free extract adsorbed on nano $TiO_2$ particles) inside an innovative bioreactor containing capsules of $N_2$ and $H_2$. In addition, the effects of different $N_2$ and $H_2$ treatments on the formation of $NH_3$ were assayed. The results showed that the dry biomass of the strains produced the most $NH_3$. The dry biomass of the Paenibacillus strain E produced the most $NH_3$ at 1.50, 0.34, and 0.27 ${\mu}M$ $NH_3$/mg biomass/h in the presence of $N_2$ + $H_2$, $N_2$, and $H_2$, respectively, indicating that a combined effluent of $N_2$ and $H_2$ was vital for $NH_3$ production. Notwithstanding, a cell-free extract (CFE) adsorbed on nano $TiO_2$ particles produced the most $NH_3$ and preserved the enzyme activities for a longer period of time, where the $NH_3$ production was 2.45 ${\mu}M$/mg CFE/h over 17 h. Therefore, the present study provides a new, simple, and inexpensive method of $NH_3$ production.

• Journal of Microbiology and Biotechnology
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• v.24 no.5
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• pp.619-628
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• 2014

To identify lipase LipA (PFL_0617) from Pseudomonas protegens Pf-5, a lipA deletion mutant (Pf0617) and a complementary strain (Pf0617lipA) were constructed, and their effects on the lipase production were examined. Pf0617 remarkably decreased its whole-cell lipase activity, whereas Pf0617lipA made its whole-cell lipase activity not only restore to wild-type level but also get a further increment. However, the deletion and overexpression of lipA did not affect the extracellular lipase activity. In addition, the unbroken whole cells of these strains were able to catalyze the hydrolysis of membrane-permeable p-nitrophenyl esters, but could not hydrolyze the membrane-impermeable olive oil. These results confirmed that LipA was an intracellular lipase and Pf-5 could also be used as a natural whole-cell biocatalyst. To evaluate the potential of Pf-5 as a whole-cell biocatalyst and separately characterize the whole-cell LipA, the properties of the whole-cell lipases from Pf-5 and Top10lipA were characterized. The results demonstrated that both Pf-5 and Top10lipA exhibited high tolerance to alkaline condition, high temperature, heavy metal ions, surfactants, and organic solvents. Taken together, lipA can realize functional expression in E. coli Top10, and Pf-5 and Top10lipA as whole-cell biocatalysts may have enormous potential in applications.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.171-178
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• 2017

A series of novel effector molecules secreted by the type three secretion system (T3SS) of Shigella spp. have been reported in recent years. In this study, a proteomic approach was applied to study T3SS effectors systematically. First, proteins secreted by the S. flexneri wild-type strain after Congo Red induction were separated and identified using two-dimensional electrophoresis to display the relative abundance of all kinds of early effectors for the first time. Then, a gene deletion mutant of known virulence repressor (OspD1) and a gene overexpressed mutant of two known virulence activators (MxiE and IpgC) were constructed and analyzed to discover potential late effectors. Furthermore, the supernatant proteins of gene deletion mutants of two known translocators (IpaB and IpaD), which would constantly secrete effectors, were also analyzed. Among all of the secreted proteins identified in our study, IpaH1.4, IpaH_5, and IpaH_7 have not been reported before. These proteomics data of the secreted effectors will be valuable to understand the pathogenesis of S. flexneri.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.563-569
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• 2004

The DNA sequence of the chitosanase gene (choK) from $\beta$-Proteobacterium KNU3 showed an 1,158-bp open reading frame that encodes a protein of 386 amino acids with a novel 74 signal peptide. The degenerated primers based on the partial deduced amino acid sequences from MALDI- TOF MS analyses yielded the 820 bp of the PCR product. Based on this information, double inverse PCR cloning experiments, which use the two specific sets of PCR primers rather than single set primers, identified the unknown 1.2 kb of the choK gene. Subsequently, a 1.8 kb of full choK gene was cloned from another PCR cloning experiment and it was then subcloned into pGEM T-easy and pUC18 vectors. The recombinant E. coli clone harboring recombinant pUC18 vector produced a clear halo around the colony in the glycol chitosan plates. The recombinant ChoK protein was secreted into medium in a mature form while the intracellular ChoK was produced without signal peptide cleavage. The activity staining of PAGE showed that the recombinant ChoK protein was identical to the chitosanase of wild-type. The comparison of deduced amino acid sequences of choK revealed that there is 92% identity with that of Sphingobacterium multivorum chitosanase. Judging from the conserved module in other bacterial chitosanases, chitosanase of KNU3 strain (ChoK) belongs to the family 80 of glycoside hydrolases.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1061-1065
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• 2001

The antibacterial effects of water extracts of Scutellariate Radix (a dried root of Scutellaria baicalensis GEORGI) and its major flavonoid components, Baicalin and Baicalein, on Salmonella typhimurium, a representative enteric pathogen, were studied. Through a Kriby-Bauer disc analysis, the growth-inhibition activity of Scutellariae Radix against. S. typhimurium was found to be compatible with commercial antibiotics, such as ampicillin, chloramphenicol, and streptomycin. In contrast, the growth of a nonpathogenic E. coli strain was unaffercted by Scutellariae Radix. To examine the effect of polyphosphate kinase (ppk), a putative virulence factor, on the antibacterial activity of Scutellariae Radix, the growth profile of a ppk mutant of S. typhimurium was investigated in a tryptic soy broth containing different concentrations of water extracts of Scutellariae Radix. The ppk mutant was able to grow in 6 mg/ml of water extracts of Scutellariae Radix, whereas in 6 mg/ml of water extracts of Scutellariae Radix, whereas the wild-type could not, implying that the inactivation of ppk made S. typhimurium more resistant to the antibacterial activity of Scutellariae Radix. No enhanced resistance was observed in a ppk mutant of S. typhimurium complemented with a ppk expression vector. The attenuation of the virulence by ppk inactivation was also observed in a virulence assay using BLAB/c mice. Neither Baicalin nor Baicalein exhibited any growth-inhibition activity against S. typhimurium. The water extracts of Scutellariae Radix stimulated the transcription of ppk, especially in the early growth-stage of S. typhimurium.

• Microbiology and Biotechnology Letters
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• v.44 no.1
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• pp.34-39
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• 2016

Wild-type yeast strains were isolated from nuruk, a type of microbial starter culture used for fermenting grains to produce alcoholic products, that was collected from different areas in Korea. Strains were identified based on the analysis of 18S rRNA sequences. Fifty strains shared the highest sequence similarity with Saccharomyces cerevisiae and were designated MBYK1-MBYK50. Among these S. cerevisiae isolates, MBYK45 produced $44.0{\pm}0.3g$ of ethanol from 200 g maltose after incubation at $30^{\circ}C$ for 48 h. Maximum ethanol production of $110.80{\pm}0.81g/l$ with productivity of $3.79{\pm}0.14g^{-1}l^{-1}h^{-1}$ was obtained at optimum culture conditions of pH (6.0), maltose (200 g/l), and temperature ($35^{\circ}C$). This study indicates that the MBYK45 strain of S. cerevisiae, isolated from nuruk, might be suitable for traditional liquor production from malts.