• The Plant Pathology Journal
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• v.16 no.4
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• pp.200-205
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• 2000

Biological control of blue stain fungi, such as Ophiostoma and Leptographium spp., that reduce the quality of logs and cause economic losses in wood product industry, was carried out in laboratory and field trials by a colorless strain of Ophiostoma quercus, BSFcs-1. Inoculation of pine wood chips with the colorless strain 1 wk before inoculating wild-type strain demonstrated that BSFcs-1 colonized wood chips and excluded blue stain fungi from being established. Efficacy of BSFcs-1 was compared with colorless strain of O. piliferum, which is commercially available under the trade name of Cartapip. Inoculation of pine wood logs with the colorless strain 1 wk before inoculating wild-type strain of blue stain in isolated wood chips, while O. quercus and O. floccosum colonized 0% and 17%, respectively. Simultaneous inoculation of logs with the colorless and wild-type strains resulted in decreased colonization (28%) by BSFcs-1, but increased colonization by O. quercus (185) and O. floccosum (29%). On the other hand, BSFcs-1 and wild-type strain alone colonized 75% and 71%, respectively. Treatment of the surface of log ends with mycelial suspension of BSFcs-1 after cutting also showed good control of blue stain fungi in a pine forest stands.

• Journal of Microbiology
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• v.42 no.3
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• pp.174-180
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• 2004

A halophilic bacterium was isolated from fermented seafood. The 16S rDNA sequence identity between the isolate and Halomonas subglaciescola AJ306801 was above 95％. The isolate that did not grow in the condition without NaCl or in the condition with other sodium (Na$\^$+/) or chloride ions (Cl$\^$-/) instead of NaCl was named H. subglaciescola DH-l. Two mutants capable of growing without NaCl were obtained by random mutagenesis, of which their total soluble protein profiles were compared with those of the wild type by two-dimensional electrophoresis. The external compatible solutes (betaine and choline) and cell extract of the wild type did not function as osmoprotectants, and these parameters within the mutants did not enhance their growth in the saline environment. In the proton translocation test, rapid acidification of the reactant was not detected for the wild type, but it was detected for the mutant in the condition without NaCl. From these results, we derived the hypothesis that NaCl may be absolutely required for the energy metabolism of H. subglaciescola DH-l but not for its osmoregulation, and the mutants may have another modified proton translocation system that is independent of NaCl, except for those mutants with an NaCl-dependent system.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.242-248
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• 2002

Previously, it was reported that the nonhomologous C-terminal regions of the D-hydantoinases are nonessential for catalysis, but affect the oligomeric structure of the enzyme [3]. In an effort to further confirm the above observation, the C-terminal region-inserted enzyme was constructed by attaching a peptide (22 residues) at the C-terminal of the D-hydantoinase from Bacillus thermocatenulatus GH2, and its structural and biochemical properties were compared with both the wild-type and C-terminal region-truncated enzymes. As a result, native tetrameric D-hydantoinase was dimerized as the truncated enzyme, and the inserted mutant with a new sequence was expressed as a catalytically active form in E. coli. Expression level of the inserted and truncated enzymes were found to be significantly increased compared to the level of the wild-type enzyme, and this appears to be due to the reduced toxic effect of the mutant enzymes on host cells. Dimerized enzymes exhibited increased thermo- and pH stabilities considerably when compared with the corresponding wild-type enzyme. Comparison of the substrate specificity between the mutant and wild-type enzymes suggests that the substrate specificity of the D-hydantoinase is closely linked with the oligomeric structure.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1871-1879
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• 2015

The complex roles of flagella in the pathogenesis of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are important. Compared with the wild-type, an insertional mutation of the flgA gene (cj0769c) demonstrated significant decrease in the biofilm formation of C. jejuni NCTC11168 on major food contact surfaces, such as polystyrene, stainless steel, and borosilicate glass. The flgA mutant was completely devoid of flagella and non-motile whereas the wild-type displayed the full-length flagella and motility. In addition, the biofilm formation of the wild-type was inversely dependent on the viscosity of the media. These results support that flagellar-mediated motility plays a significant role in the biofilm formation of C. jejuni NCTC11168. Moreover, our adhesion assay suggests that it plays an important role during biofilm maturation after initial attachment. Furthermore, C. jejuni NCTC11168 wild-type formed biofilm with a net-like structure of extracellular fiber-like material, but such a structure was significantly reduced in the biofilm of the flgA mutant. It supports that the extracellular fiber-like material may play a significant role in the biofilm formation of C. jejuni. This study demonstrated that flgA is essential for flagellar biosynthesis and motility, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.

• Journal of Microbiology and Biotechnology
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• v.9 no.1
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• pp.84-90
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• 1999

The growth of Bifidobacterium breve strain HP2 was completely inhibited by the addition of lactate higher than 4.0％ but not by the addition of acetate. Two kinds of lactate-tolerant mutants were isolated by the nitrosoguanidine treatment, enrichment on a liquid medium with 5％ lactate, and selection on agar plates with 5％ lactate. The mutants were not only able to grow in the presence of 5％ lactate but also improved in viable cell stability in the acidic pH range. In a pH-controlled fermentor, mutant N-1-5 grew at a rate slower than that of the wild type but its growth yield was higher. Notably, mutants were more halotolerant and more osmotolerant than the wild type and they were able to grow in the presence of 3％ NaCl or 25％ lactose at which the wild type entirely stopped the growth. The enzyme activities involved in the lactose metabolism in B. breve were measured to elucidate the biochemical basis for lactate tolerance. In the mutants, activities of several enzymes including phosphoglucomutase decreased compared to the wild-type, which may explain their lower growth rate. However, the activity of lactate dehydrogenase or its nature of inhibition by lactate was not altered.

• BMB Reports
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• v.40 no.3
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• pp.333-340
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• 2007

Gene encoding cyclodextrin glycosyltransferase (CGTase), from thermotolerant Paenibacillus sp. T16 isolated from hot spring area in northern Thailand, was cloned and expressed in E. coli (JM109). The nucleotide sequences of both wild type and transformed CGTases consisted of 2139 bp open reading frame, 713 deduced amino acids residues with difference of 4 amino acid residues. The recombinant cells required 24 h culture time and a neutral pH for culture medium to produce compatible amount of CGTase compared to 72 h culture time and pH 10 for wild type. The recombinant and wild-type CGTases were purified by starch adsorption and phenyl sepharose column chromatography and characterized in parallel. Both enzymes showed molecular weight of 77 kDa and similar optimum pHs and temperatures with recombinant enzyme showing broader range. There were some significant difference in pH, temperature stability and kinetic parameters. The presence of high starch concentration resulted in higher thermostability in recombinant enzyme than the wild type. The recombinant enzyme was more stable at higher temperature and lower pH, with lower $K_m$ for coupling reaction using cellobiose and cyclodextrins as substrates.

• Asian Journal of Turfgrass Science
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• v.11 no.1
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• pp.45-58
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• 1997

Pigments and thylakoid membrane proteins were investigated in wild type and PS I- mutants from Synechocystis sp. PCC6803 Comparing morphological features, B2 was less fluorescent than the other strains. The contents of chlorophyll a were propotional to the FNR activity in thylakoid membrane. The FNR activity of mutants was lower than that of wild type. In the result of pigments analysis, mutants had smaller cholophyll a than that of wild type. The major carotenoid was found to he $\beta$-caroene, but aeaxanthin was barely detected in thylakoid membrane of mutants. The polypeptide, 14.8kD was detected by electrophoresis in mutants. It was considered to be the modification of 15.4kD in wild type. Membrane polypeptides of 17.6 and 19.7kD were not detected in mutants. In the result of western blotting, subunit I was detected in all strains, but subunit II was barely detected in mutants. Subunit II was not detected in B2 at all. In view of the results so far achieved, the changes of contents of chlorophyll and zeaxanthin were affected by the defficiency or modification of functional domain in subunit I. Also the modification in subunit I affected the subunit II- binding site in PS I. As the result, efficiency of photosynthesis was decreased. Key words: Synechoystis sp. PCC6803, PS I - mutant, Photosynthetic efficiency, Pigment,Thylakoid membrane proteins, Subunit I, II.

• Microbiology and Biotechnology Letters
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• v.22 no.5
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• pp.538-543
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• 1994

Effect of partial oxygen pressure on the cell growth and the activities of oxidative defense enzymes were measured in the continuous culture of Streptomyces coelicolor. Both the wild type and the mutant strain resistant to hydrogen peroxide were cultured and the dry cell weight of the two cultures were measured at different oxygen tensions. Growth of the wild type was inhibited by oxygen at above 0.5 vvm. Growth of the hydrogen peroxide resistant mutant was stimulated by pure oxygen at 0.5 vvm but was inhibited by oxygen at 1.0 vvm. Therefore, growth of the hydrogen peroxide resistant mutant was less affected by the deleterious oxidative stress of oxygen. Activities of the several defense enzymes were also measured at different oxygen tensions. Activities of catalase and glucose-6-phosphate dehydrogenase increased significantly as oxygen pressure increased in the wild type culture. In the mutant, however, increase in those enzyme activities was not obvious whereas the uninduced levels of the above enzymes were higher than those of wild type. As judged by Western blotting, the amount of the major catalase increased as the oxygen pressure increased. This indicates that the induction of the catalase activity by oxygen pressure is mostly due to the increase in the expression level for the major catalase.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1066-1070
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• 2001

The PhyA gene, encoding myo-inositol hexakisphosphate phosphohydrolase in Aspergillus niger var. awamori (wild-type), was cloned and sequenced. The cDNA was overexpressed by a multicopy gene expression system in Pichia pastoris KM71. Recombinant, wild-type and commercial phytase from Aspergilus ficuum NRRL 3135 (Natuphos) were purified. The PhyA gene of Aspergillus niger var awamori showed perfect homology to the phytase of Aspergillus ficcum and $97\%$ homology to A. niger var awamori (L02421). Wild-type phytase was highly glycosylated and more thermostable than the other two, while deglycosylated farms of three phytases showed identical molecular weight, 507 kDa. After heating at $80^{\circ}C$, wild-type, commercial, and recombinant phytases retained $57\%, 32%,\;and\;8\%$ of their original activities, respectively. In conclusion, glycosylation plays a key role in the thermostability of phytase and its enzymatic characterization.

• Journal of Microbiology and Biotechnology
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• v.19 no.4
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• pp.424-430
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• 2009

IL-10 is an important anti-inflammatory cytokine that can inhibit the production of many pro-inflammatory cytokines. Both human and animal studies have shown that pro-inflammatory cytokines play an important role in pneumonia and other inflammatory lung diseases. In the present study, IL-10 knockout(KO) and wild-type mice were infected with Corynebacterium kutscheri to determine whether the severity of pathogenesis and whether protective immunity could be altered in the absence of IL-10. The survival rate was significantly lower in IL-10 KO mice than wild-type mice. The number of neutrophils in bronchoalveolar lavage fluid and blood were found to be higher in IL-10 KO mice than wild-type mice. IL-10 KO mice showed greater neutrophil infiltration, excessive inflammation, and weight-loss compared with wild-type mice. Furthermore, upregulation of IFN-$\gamma$ in bronchoalveolar lavage fluid, and upregulation of MIP-$1{\alpha}$ and IP-10 mRNA in the lungs of IL-10 KO mice compared with wild-type mice after C. kutscheri infection were observed. These results suggest that IL-10 plays an important role in the anti-inflammatory properties against C. kutscheri infection, and that lack of IL-10 leads to a more severe pulmonary inflammatory response. This increased susceptibility to C. kutscheri pneumonia is at least in part caused by IL-10 deficiency and severe recruitment of neutrophils.