• Title/Summary/Keyword: wild strain

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Expression of Mosquitocidal Bacillus sphaericus Binary Toxin and B. thuringiensis cry11B Genes in B. thuringiensis 407

  • Park, Hyun-Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.2
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    • pp.185-189
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    • 2001
  • Wild type Bacilus thuringiensis subsp. israelensis and B. sphaericus toxins have been used separately as active in ingredients for bacterial insecticides to control mosquito larvae due to their comparable toxicity to chemical insecticides. Cry11B, recently cloned from B. thuringiensis subsp. jegathesan, shows higher toxicity against three major species of mosquito larvae than Cry11A, one of the major component of B. thuringiensis subsp. israelensis inclusion body. To determine whether the combination of cry11B and B. sphaericus binary toxins is as toxic as B. thuringiensis subsp. israelensis parental strain, cry11B and B. sphaericus binary toxins genes were co-expressed as an operon using cytlA promoters/STAB-SD hybrid expression system in B. thuringiensis subsp. israelensis acrystalliferous strain 4Q7. However, unexpectedly, B. sphaericus binary toxins were barely produced, whereas relatively large amount of Cry11B was produced. When this strain was grown in four different media, NB+G and Peptonized Milk produced more toxin proteins and spores per unit of media than GYS and G-Tris. Toxicity of this strain against fourth instar Culex quinquefasciatus was ranged from of 8.3 to 45.7 ng/ml, with NB+G culture being the highest, and GYS culture was the lowest.

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Strain Improvement by Overexpression of the laeA Gene in Monascus pilosus for the Production of Monascus-Fermented Rice

  • Lee, Sang Sub;Lee, Jin Hee;Lee, Inhyung
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.959-965
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    • 2013
  • Monascus species have been used to produce fermented rice called Monascus-fermented rice (MFR). To improve a Monascus strain via activation of secondary metabolite (SM) gene clusters for use in the production of MFR, we overexpressed an ortholog of the laeA gene, which encodes a global positive regulator of secondary metabolism under the control of the strong heterologous Aspergillus nidulans alcA promoter in Monascus pilosus. The OE::laeA transformant produced more SMs, including those not detected under uninduced conditions. MFR produced using the M. pilosus OE::laeA strain contained 4 times more monacolin K, a cholesterol-lowering agent, than MFR produced using the wild-type strain. In addition, pigment production was remarkably increased, and the antioxidant activity was increased as well. The results from this study suggest that Monascus species, which are important industrial fermentative fungi in Asia, can be improved for the production of functional foods by overexpressing the laeA gene.

Clinical profile of Asian and African strains of Zika virus in immunocompetent mice

  • Shin, Minna;Kim, Jini;Park, Jeongho;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.61 no.2
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    • pp.12.1-12.9
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    • 2021
  • The mosquito-borne pathogen Zika virus may result in neurological disorders such as Guillain-Barré syndrome and microcephaly. The virus is classified as a member of the Flaviviridae family and its wide spread in multiple continents is a significant threat to public health. So, there is a need to develop animal models to examine the pathogenesis of the disease and to develop vaccines. To examine the clinical profile during Zika virus infection, we infected neonatal and adult wild-type mice (C57BL/6 and Balb/c) and compared the clinical signs of African-lineage strain (MR766) and Asian-lineage strain (PRVABC59, MEX2-81) of Zika virus. Consistent with previous reports, eight-week-old female Balb/c mice infected with these viral strains showed no changes in body weight, survival rate, and neurologic signs, but demonstrated increases in the weights of spleens and hearts. However, one-day-old neonates showed significantly lower survival rate and body weight with the African-lineage strain than the Asian-lineage strain. These results confirmed the pathogenic differences between Zika virus strains. We also evaluated the clinical responses in neonatal and adult mice of different strains. Our findings suggest that these are useful mouse models for characterization of Zika virus for vaccine development.

Three Non-Aspartate Amino Acid Mutations in the ComA Response Regulator Receiver Motif Severely Decrease Surfactin Production, Competence Development, and Spore Formation in Bacillus subtilis

  • Wang, Xiaoyu;Luo, Chuping;Liu, Youzhou;Nie, Yafeng;Liu, Yongfeng;Zhang, Rongsheng;Chen, Zhiyi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.301-310
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    • 2010
  • Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

Improvement of the Phosphate Solubilization Microorganism by the Introduction of Glucose Dehydrogenase Gene into Aeromonas hydrophila DA33. (Glucose dehydrogenase 유전자의 Aeromonas hydrophila DA33으로의 도입에 따른 인산가용화 균주의 개량)

  • Park, In-Hye;Song, Ok-Ryul;Lee, Yong-Seok;Kang, Ui-Gum;Choi, Si-Lim;Choi, Yong-Lark
    • Journal of Life Science
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    • v.18 no.6
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    • pp.878-883
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    • 2008
  • Aeromonas hydrophila DA33 was isolated from cultivated soils as a bacteria having high abilities to solubilize inorganic phosphate. Glucose dehydrogenase gene (gdh) was cloned from Escherichia coli. The recombinant plasmid, pGHS containing glucose dehydrogenase gene was introduced into A. hydrophila DA33 in order to improve the activity of phosphate-solubilizing. The transformant harboring the gdh gene, A. hydrophila pGHS/DA33 increased enzyme activity. The strain also increased the gluconic acid generation that was effective for phosphate solubilization. It was possible that the strain containing pGHS produced higher solubilized phosphate with tri-calcium phosphate as the unique (P) source, in comparison with that of wild type without plasmid. These results suggest that the strain, A. hydrophila pGHS/DA33 is expected as effective biofertilizer for phosphate solubilization.

Downregulation of EHT1 and EEB1 in Saccharomyces cerevisiae Alters the Ester Profile of Wine during Fermentation

  • Yang, Xue;Zhang, Xuenan;He, Xi;Liu, Canzhen;Zhao, Xinjie;Han, Ning
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.761-767
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    • 2022
  • EHT1 and EEB1 are the key Saccharomyces cerevisiae genes involved in the synthesis of ethyl esters during wine fermentation. We constructed single (Δeht1, Δeeb1) and double (Δeht1Δeeb1) heterogenous mutant strains of the industrial diploid wine yeast EC1118 by disrupting one allele of EHT1 and/or EEB1. In addition, the aromatic profile of wine produced during fermentation of simulated grape juice by these mutant strains was also analyzed. The expression levels of EHT1 and/or EEB1 in the relevant mutants were less than 50% of the wild-type strain when grown in YPD medium and simulated grape juice medium. Compared to the wild-type strain, all mutants produced lower amounts of ethyl esters in the fermented grape juice and also resulted in distinct ethyl ester profiles. ATF2, a gene involved in acetate ester synthesis, was expressed at higher levels in the EEB1 downregulation mutants compared to the wild-type and Δeht1 strains during fermentation, which was consistent with the content of acetate esters. In addition, the production of higher alcohols was also markedly affected by the decrease in EEB1 levels. Compared to EHT1, EEB1 downregulation had a greater impact on the production of acetate esters and higher alcohols, suggesting that controlling EEB1 expression could be an effective means to regulate the content of these aromatic metabolites in wine. Taken together, the synthesis of ethyl esters can be decreased by deleting one allele of EHT1 and EEB1 in the diploid EC1118 strain, which may modify the ester profile of wine more subtly compared to the complete deletion of target genes.

Analysis of physiological activities of wild Flammulina velutipes (야생 팽나무버섯의 생리활성 분석)

  • Yoon, Hyang-Sik;Choi, Jae-Sun;Kim, Ki-Sik;Noh, Jae Goan;Park, Jae Seong;Rho, Chang Woo
    • Journal of Mushroom
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    • v.8 no.1
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    • pp.22-26
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    • 2010
  • In this study, for the development of useful food materials, the proximate composition(total carbohydrates, crude fiber, crude protein, fat and ash) and biological activities(electron donating activity, total polyphenol contents, ACE inhibitory and ${\gamma}$-glucosidase inhibitory effects) were determined in the Garlmoe and the four wild Flammulina velutipes. As a results, the dry matter of mushrooms contained large amounts of carbohydrates, from 65.9 to 70.5 g/100g dry weight. the wild collected Flammulina velutipes proved to be an especially good source of crude fiber(7.9~18.1 g/100 g dry weight). Crude protein, fat, and ash contents of the mushrooms ranged 17.1~21.3, 4.1~5.2, and 5.7~8.4 g/100g dry weight, respectively. The electron donating activity ranged from 63.35 to 69.31% and total polyphenol contents ranged from 9.83 to 11.14 mg/g in these mushrooms. ACE inhibitions were high in Garlmoe and CBMFV-02 strain. ${\alpha}$-glucosidase inhibition was the highest in CBMFV-01(46.56%) strain and lowest in Garlmoe(15.91%).

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Arthrospira platensis Mutants Containing High Lipid Content by Electron Beam Irradiation and Analysis of Its Fatty Acid Composition (전자빔 조사에 의해 지질 함량이 증대된 Arthrospira platensis 변이주 분리 및 지방산 분석)

  • Choi, Soo-Jeong;Kim, Young-Hwa;Kim, Andre;Lee, Jae-Hwa
    • Applied Chemistry for Engineering
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    • v.24 no.6
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    • pp.628-632
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    • 2013
  • Arthrospira platensis (A. platensis) is an economically important microalgae because it has carbohydrates, lipids, proteins and a number of phytochemicals. It is also a valuable source used in the production of biodiesel and functional foods. In this study, A. platensis was exposed to electron beam irradation (240 kGy) and induced random mutagenesis for strain improvement. Several mutants were obtained, and the resulting mutant was designated as EB29. The growth rate and chlorophyll content of EB29 was similar to those of wild type. However, the lipid content of EB29 was increased seven-fold compared to that of wild type when comparing the nile red fluorescent intensity. Semi-quantitative analysis of EB29 using the calibration plot of standard lipid, triolein, represented $78.6{\mu}g/mL$, which increased 2 times compared to wild type ($41.4{\mu}g/mL$). When analyzing the fatty acid profile of EB29, polyunsaturated fatty acids (PUFAs), such as gamma-linolenic acid (GLA) in EB29 increased about six-fold. Moreover, fatty acids affecting the quality of biodiesel increased compared to that of wild type. Thus, electron beam could be used for the strain improvement of microalgae in order to accumulate PUFAs and alteration of fatty acid profile for biodiesel.

Eritadenin Contents Analysis in Various Strains of Lentinula edodes using LC-MS/MS (LC-MS/MS를 이용한 표고 균주별 에리타데닌 함량 분석)

  • Park, Young-Ae;Lee, Kyoung-Tae;Bak, Won-Chull;Kim, Myung-Kil;Ka, Kang-Hyeon;Koo, Chung-Duck
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.239-242
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    • 2011
  • Eritadenine, a potent compound of hypocholesterolemic activity, was investigated in relation to its content in domestic cultivars and wild types of shiitake (Lentinula edodes). Eighteens strains of shiitake were tested for the quantification of eritadenine by LC-MS/MS analysis. Among the strains, wild type-40 was highest as the content was 3.912 mg/g. Also, Soohyangko was 3.352mg/g, Sanlim No. 9 3.008mg/g, Chunbaegko 2.832 mg/g, Gaeulhyang and KFRI 675 both 2.792 mg/g as high-content strains. Soohyangko and Chunbaegko are applied strains for registration in 2010. Soohyangko is high-temperature type with concentrated fruiting, and 90% of production occurs in the first year, thus, recovery of cost is very fast. Chunbaegko is mid-temperature type with concentrated flushing, and produces "hwago", the best quality, in spring. Wild type-40 is excellent in productivity and is prepared for registration. Wild type-40 could be used as parent strain to make new strain with high eritadenine content.

Immune-Enhancing Alkali-Soluble Glucans Produced by Wild-Type and Mutant Saccharomyces cerevisiae

  • Ha Chang-Hoon;Lim Ki-Hong;Jang Se-Hwan;Yun Cheol-Won;Paik Hyun-Dong;Kim Seung-Wook;Kang Chang-Won;Chang Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.576-583
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    • 2006
  • The alkali-soluble glucan of the yeast cell wall contains $\beta-(1,3)-$ and (1,6)-D-linkages and is known to systemically enhance the immune system. In the previous study [6], in order to isolate cell wall mutants, a wild-type strain was mutagenized by exposure to ultraviolet light, and the mutants were then selected via treatment with laminarinase $(endo-\beta-(1,3)-D-glucanase)$. The mass of alkali- and water-soluble glucans produced by the mutant was measured to be 33.8 mg/g of the dry mass of the yeast cell. Our results showed that the mutants generated the amount of alkali-soluble glucan 10-fold higher than that generated by the wild-type. Structural analysis showed that the alkali-soluble glucan from the mutants was associated with a higher degree of $\beta-(1,6)-D-linkage$ than was observed in conjunction with the wild-type. Yeast cell wall $\beta-glucan$ was shown to interact with macrophages via receptors, thereby inducing the release of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide. Alkali-soluble $\beta-glucans$, both from water-soluble and water-insoluble glucan, exhibited a higher degree of macrophage activity with regard to both the secretion of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide and direct phagocytosis, than did the positive control ($1{\mu}g$ of lipopolysaccharide).