• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.360-367
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• 2006

The endoinulinase gene (inu1) from Pseudomonas mucidolens was expressed on the cell surface of Saccharomyces cerevisiae by fusing with Aga2p linked to the membrane anchored protein, Aga1p. The inu1 gene of P. mucidolens was subcloned into the surface display vector, pCTcon (GAL1 promoter). The constructed plasmid, pCTENIU (8.5kb), was then introduced to S. cerevisiae EBY100 cells and the yeast transformants selected on synthetic defined media lacking uracil and inulin-containing media. The inu1 gene under the control of the GAL1 promoter was successfully expressed in the yeast transformants, and the surface display of endoinulinase confirmed by immunofluorescence microscopy, along with its enzymatic ability to form inulooligosaccharides (IOSs) from inulin. The total endoinulinase activity reached about 2.31 units/ml when the yeast transform ants were cultivated on a YPDG medium. To efficiently hydrolyze the inulin, various reaction conditions were examined, including the pH, temperature, and inulin source. The optimized conditions were then determined as follows: pH, 7.0; temperature, $50^{\circ}C$; inulin source, Jerusalem artichoke. Under the optimized condition and 46 units of endoinulinase per g of inulin, IOSs started to be produced after 10 min of enzymatic reaction. The highest yield, 71.2% of IOSs, was achieved after 30 h of reaction without any significant loss of the initial enzyme activity. As a result of the reaction with inulin, IOSs consisting of inulobiose (F2), inulotriose (F3), inulotetraose (F4), and inulopentaose (F5) were produced, and F4 was the major product.

• 한국환경농학회지
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• 제25권2호
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• pp.124-128
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• 2006

황 화합물 악취를 제거하기 위하여 UV-B를 돌연변이원으로 처리한 후 공시균주 Thiobacillus sp. IW보다 성장속도가 빠른 변이주 UIW-6를 분리하여 최적성장 조건을 조사하였다. 변이주 UIW-6의 특성을 조사한 결과 최적조건은 pH 6.5, 배양온도는 $35^{\circ}C$이었으며, 회전속도는 200 rpm 이었다. 황 화합물인 sodium thiosulfate 농도가 50 mM에서 변이주 UIW-6는 6시간째에 공시균주 IW보다 성장정도가 2배 더 빨랐다. 변이주 UIW-6의 성장 최적 탄소원은 fructose와 sucrose였으며 질소원은 yeast extract> tryptone> peptone 순으로 이용하였다. 변이주 UIW-6는 공시균주로 사용한 IW 보다 성장이 빨라 황 화합물 악취 제거에 효과적으로 이용할 수 있는 유용한 변이주로 판단되어진다.

• Mycobiology
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• 제42권4호
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• pp.368-375
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• 2014

Red ginseng (Panax ginseng), a Korean traditional medicinal plant, contains a variety of ginsenosides as major functional components. It is necessary to remove sugar moieties from the major ginsenosides, which have a lower absorption rate into the intestine, to obtain the aglycone form. To screen for microorganisms showing bioconversion activity for ginsenosides from red ginseng, 50 yeast strains were isolated from Korean traditional meju (a starter culture made with soybean and wheat flour for the fermentation of soybean paste). Twenty strains in which a black zone formed around the colony on esculin-yeast malt agar plates were screened first, and among them 5 strains having high ${\beta}$-glucosidase activity on p-nitrophenyl-${\beta}$-D-glucopyranoside as a substrate were then selected. Strain JNO301 was finally chosen as a bioconverting strain in this study on the basis of its high bioconversion activity for red ginseng extract as determined by thin-layer chromatography (TLC) analysis. The selected bioconversion strain was identified as Candida allociferrii JNO301 based on the nucleotide sequence analysis of the 18S rRNA gene. The optimum temperature and pH for the cell growth were $20{\sim}30^{\circ}C$ and pH 5~8, respectively. TLC analysis confirmed that C. allociferrii JNO301 converted ginsenoside Rb1 into Rd and then into F2, Rb2 into compound O, Rc into compound Mc1, and Rf into Rh1. Quantitative analysis using high-performance liquid chromatography showed that bioconversion of red ginseng extract resulted in an increase of 2.73, 3.32, 33.87, 16, and 5.48 fold in the concentration of Rd, F2, compound O, compound Mc1, and Rh1, respectively.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.917-930
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• 2018

Intracellular synthesis of silver/silver chloride nanoparticles (Ag/AgCl-NPs) using Meyerozyma guilliermondii KX008616 is reported under aerobic and anaerobic conditions for the first time. The biogenic synthesis of Ag-NP types has been proposed as an easy and cost-effective alternative for various biomedical applications. The interaction of nanoparticles with ethanol production was mentioned. The purified biogenic Ag/AgCl-nanoparticles were characterized by different spectroscopic and microscopic approaches. The purified nanoparticles exhibited a surface plasmon resonance band at 419 and 415 nm, confirming the formation of Ag/AgCl-NPs under aerobic and anaerobic conditions, respectively. The planes of the cubic crystalline phase of the Ag/AgCl-NPs were confirmed by X-ray diffraction. Fourier-transform infrared spectra showed the interactions between the yeast cell constituents and silver ions to form the biogenic Ag/AgCl-NPs. The intracellular Ag/AgCl-NPs synthesized under aerobic condition were homogenous and spherical in shape, with an approximate particle size of 2.5-30nm as denoted by the transmission electron microscopy (TEM). The reaction mixture was optimized by varying reaction parameters, including temperature and pH. Analysis of ultrathin sections of yeast cells by TEM indicated that the biogenic nanoparticles were formed as clusters, known as nanoaggregates, in the cytoplasm or in the inner and outer regions of the cell wall. The study recommends using the biomass of yeast that is used in industrial or fermentation purposes to produce Ag/AgCl-NPs as associated by-products to maximize benefit and to reduce the production cost.

• Archives of Pharmacal Research
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• 제12권2호
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• pp.79-87
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• 1989

Enzymatic methylation of arginine and lysine residues of several cytochrome c and lysine residue of calmodulin always resulted in lowering of their respective isoelectric points (pI). Employing cytochromes c derived from various sources, we examined a possible relationship between the degree of amino acid sequence degeneracy and the magnitude of change in the pI values by enzymatic methylation, and found that there was no correlation between these two parameters. By constructing space-filling models of oligopeptide fragments adjacent to the potential methylation sites, we have noted that not all the methylatable residues are able to form hydrogen bonds prior to the methylation. Two preparations of yeast apocytochrome c, one chemically prepared by removing heme from holocytochrome c and the other by translating yeast iso-1-cytochrome c mRNA in vitro, exhibited slightly higher Stokes radii than the homologous holocytochrome c, indicating relatively 'relaxed or open' conformation of the protein. However, when the in vitro synthesized methylated apocytochrome c was compared with the unmethylated counter-part, the Stokes radius of the latter was found to be larger.

• Journal of Microbiology and Biotechnology
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• 제10권4호
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• pp.482-487
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• 2000

Optimization of submerged culture conditions for the production of exo-biopolymer from Paecilomyces japonica ws studied. Maltose, yeast extract, and potassium phosphate were the most suitable sources of carbon, nitrogen, and inorganic salt, respectively, for both production of the exo-biopolymer and mycelial growth. The optimal culture conditions in a flask culture were pH 5.0, $25^{\circ}C$, and 150 rpm in a medium containing (as in g/l) 30 maltose, 6 yeast extruct, 2 polypeptone, $0.5{\;}K_3HPO_4,{\;}0.2{\;}KH_2PO_4,{\;}0.2{\;}MnSO_4{\cdot}5H_2O,{\;}0.2{\;}MgSO_4{\cdot}7H_2O$. Exo-biopolymer production and mycelial growth in the above suggested medium were significantly increased in a 2.5-1 jar fermentor, where the maximum biopolymer concentration was 8 g/l. The morphological changes of the mycelium in the submerged culture were observed within pH ranges from 4.0 to 9.0; i.e., growth of the filamentous form was optimal at culture pHs of 5.0 and 6.0, whereas pellet was formed at other pHs.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1207-1215
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• 2014

Candida albicans, the major human fungal pathogen, undergoes morphological transition from the budding yeast form to filamentous growth in response to nitrogen starvation. In this study, we identified a new function of GST2, whose expression was required for filamentous growth of C. albicans under nitrogen-limiting conditions. The Gst2p showed Gst activity and required response to oxidative stress. The ${\Delta}gst2$ mutant displayed predominantly yeast phase growth in low ammonium media. Such morphological defect of ${\Delta}gst2$ mutants was not rescued by overexpression of Mep2p, Cph1p, or Efg1p, but was rescued by either overexpression of a hyperactive $RAS1^{G13V}$ allele or through exogenous addition of cyclic AMP. In addition, the ${\Delta}gst2$ mutants had lower levels of RAS1 transcripts than wild-type cells under conditions of nitrogen starvation. These results were consistent with the Ras1-cAMP pathway as a possible downstream target of Gst2p. These findings suggest that Gst2p is a significant component of nitrogen starvation-induced filamentation in C. albicans.

• Journal of Microbiology and Biotechnology
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• 제19권2호
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• pp.161-166
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• 2009

Certain microalgae have been known to use light and various carbon sources to produce carbohydrates, mainly in the form of starch. This is one of the pertinent feedstocks replacing agricultural products for the production of bioethanol by yeast. This study focuses upon dilute acid hydrothermal pretreatments at low cost and high efficiency to compete with current methods, and employs Chlamydomonas reinhardtii UTEX 90 as the feedstock. With dry cells of 5%(w/v), the algal biomass was pretreated with sulfuric acid(1-5%) under temperatures from 100 to $120^{\circ}C$, from 15 to 120 min. As a result, the glucose release from the biomass was maximum at 58%(w/w) after pretreatment with 3% sulfuric acid at $110^{\circ}C$ for 30 min. This method enabled not only starch, but also the hydrolysis of other oligosaccharides in the algal cell in high efficiency. Arrhenius-type of model equation enabled extrapolation of some yields of glucose beyond this range. The pretreated slurry was fermented by yeast, Saccharomyces cerevisiae S288C, resulting in an ethanol yield of 29.2% from algal biomass. This study suggests that the pretreated algal biomass is a suitable feedstock for ethanol production and can have a positive impact on large-scale applied systems.

• 대한의생명과학회지
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• 제12권4호
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• pp.405-411
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• 2006

Heterotrimeric GTP binding proteins (G proteins) mediate signal transduction generated by neurotransmitter and hormones. Among G-proteins, Go is classified as a member of the Go/Gi family and the most abundant heterotrimeric G protein in brain. Most of the mechanistic analyses on the activation of Go indicated its action to be mediated by the $G{\beta}{\gamma}$ dimer because downstream effectors for its ${\alpha}$ subunit have not been clearly defined. To determine the downstream effectors of alpha subunits of Go ($Go{\alpha}$), we used yeast two-hybrid system to screen $Go{\alpha}$ interacting partners in cDNA library from the human brain. A brain specific high mobility group box containing protein (BHX), A possible transcription factor, was identified as a $Go{\alpha}$ interacting protein. We confirmed interaction between $Go{\alpha}$ and BHX employing in vitro affinity binding assay. Moreover, active form of $Go{\alpha}$ preferentially interacts with BHX than inactive farm. Our findings indicate that $Go{\alpha}$ could modulate gene expression via interaction with BHX during neuronal or brain development.

• 한국식품과학회지
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• 제26권3호
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• pp.195-198
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• 1994

Yeast 발효공정에서 발생되는 가스량을 전자적으로 monitoring 할 수 있는 새로운 기포수 계측 쎈서를 설계, 제작하였다. 본 기포쎈서는 기체를 발생하는 발효공정에 적용할 수 있으며 공정의 상태와 환경에 아무런 변화를 주지 않도록 설계하였다. 그 구조는 가스포집부, 기포형성부, 전자식 기포감지부와 컴퓨터 접속장치로 구성하였다. 기포 계측부는 photo-interrupter카 counter IC 등을 사용하였다. 제작된 쎈서와 컴퓨터 시스템은 효모 배양공정에 활용하여 발효진행과정을 자동적으로 monitoring 하는데 성공하였으며 발효공정에서 발생된 기포수의 값으로 작성된 가스발생량 곡선은 효모의 생육곡선의 양상과 매우 유사한 모양을 보였으며 배양액의 기질영양원이 달리한 발효공정을 monitering한 결과 기질이용 특성도 정확하게 표현할 수 있었다.