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        An efficient process for co-production of γ-aminobutyric acid and probiotic Bacillus subtilis cells

        Hongbo Wang,Jinge Huang,Lei Sun,Fuchao Xu,Wei Zhang,Jixun Zhan 한국식품과학회 2019 Food Science and Biotechnology Vol.28 No.1

        This study was to establish an integrated processfor the co-production of c-aminobutyric acid (GABA) andlive probiotics. Six probiotic bacteria were screened andBacillus subtilis ATCC 6051 showed the highest GABAproducingcapacity. The optimal temperature and initial pHvalue for GABA production in B. subtilis were found to be30 C and 8.0, respectively. A variety of carbon andnitrogen sources were tested, and potato starch and peptonewere the preferred carbon and nitrogen sources for GABAproduction, respectively. The concentrations of carbonsource, nitrogen source and substrate (sodium L-glutamate)were then optimized using the response surface methodology. The GABA titer and concentration of viable cells ofB. subtilis reached 19.74 g/L and 6.0 9 108 cfu/mL at120 h. The GABA titer represents the highest production ofGABA in B. subtilis. This work thus demonstrates a highlyefficient co-production process for GABA and probiotic B. subtilis cells.

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        Identification and heterologous reconstitution of a 5-alk(en)ylresorcinol synthase from endophytic fungus Shiraia sp. Slf14

        Huiwen Yan,Lei Sun,Jinge Huang,Yixing Qiu,Fuchao Xu,Riming Yan,Du Zhu,Wei Wang,Jixun Zhan 한국미생물학회 2018 The journal of microbiology Vol.56 No.11

        A new type III polyketide synthase gene (Ssars) was discovered from the genome of Shiraia sp. Slf14, an endophytic fungal strain from Huperzia serrata. The intron-free gene was cloned from the cDNA and ligated to two expression vectors pET28a and YEpADH2p-URA3 for expression in Escherichia coli BL21(DE3) and Saccharomyces cerevisiae BJ5464, respectively. SsARS was efficiently expressed in E. coli BL21(DE3), leading to the synthesis of a series of polyketide products. Six major products were isolated from the engineered E. coli and characterized as 1,3-dihydroxyphenyl- 5-undecane, 1,3-dihydroxyphenyl-5-cis-6 -tridecene,1,3-dihydroxyphenyl- 5-tridecane, 1,3-dihydroxyphenyl-5-cis-8 - pentadecene, 1,3-dihydroxyphenyl-5-pentadecane, and 1,3- dihydroxyphenyl-5-cis-10 -heptadecene, respectively, based on the spectral data and biosynthetic origin. Expression of SsARS in the yeast also led to the synthesis of the same polyketide products, indicating that this enzyme can be reconstituted in both heterologous hosts. Supplementation of soybean oil into the culture of E. coli BL21(DE3)/SsARS increased the production titers of 1–6 and led to the synthesis of an additional product, which was identified as 5-(8 Z,11 Z-heptadecadienyl) resorcinol. This work thus allowed the identification of SsARS as a 5-alk(en)ylresorcinol synthase with flexible substrate specificity toward endogenous and exogenous fatty acids. Desired resorcinol derivatives may be synthesized by supplying corresponding fatty acids into the culture medium.

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        Overexpression of the AtSTK Gene Increases Salt, PEG and ABA Tolerance in Arabidopsis

        Lei Bing,Cui-Cui Feng,Jing-Lan Li,Xiao-Xu Li,Baocun Zhao,Yin-Zhu Shen,Zhan-Jing Huang,Rong-Chao Ge 한국식물학회 2013 Journal of Plant Biology Vol.56 No.6

        AtSTK (At5g02800), which is a serine-threonineprotein kinase gene of Arabidopsis thaliana, was cloned, andits function was studied. The study found that the overexpressionof AtSTK could significantly improve the ability of A. thaliana to tolerate salt, PEG, and ABA stresses. RT-PCRanalysis revealed that the expression of the AtSTK genecould be obviously induced by salt, PEG, and ABA. Theexamination of the physiological characteristics showed thatthe overexpression of AtSTK in Arabidopsis significantlyreduced the plasma membrane permeability, significantlyincreased the proline content, and decreased the MDA content. These changes may reflect the physiological mechanismsthrough which AtSTK overexpression improves stress resistancein Arabidopsis. In addition, the overexpression of the AtSTKgene significantly antagonised the inhibitory effect of highconcentrations of exogenous ABA on Arabidopsis seedgermination. The subcellular localisation results showed thatAtSTK is located in both the cytosol and the nucleus. Theexamination of its tissue-specific expression showed thatAtSTK is expressed in various Arabidopsis tissues and isparticularly strongly expressed in the vessels. The signallingpathway analysis indicated that AtSTK might transfer thesalt stress signal in Arabidopsis through the MAPK pathway.

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