• Title/Summary/Keyword: pH 전환

Search Result 552, Processing Time 0.028 seconds

Characterization of Alcohol Dehydrogenase Encoded by Zymomonas mobilis Gene Cloned in Escherichia coli (Escherichia coli 형질전환체가 생산하는 Zymomonas mobilis 알콜 탈수소 효소의 분석)

  • 신병식;윤기홍;박무영
    • Microbiology and Biotechnology Letters
    • /
    • v.18 no.3
    • /
    • pp.268-272
    • /
    • 1990
  • The structural gene (zadhII) encoding an alcohol dehydrogenase II from Zyrnornonas mobilis was cloned into Escherichia coli in our laboratory (Yoon et al., 1989. Kor. J. Microbiol. Biotechnol.). From E. coli (pADS93) carrying the zadhII gene, the Z mobilis alcochol dehydrogenase II (ZADH-II) was purified by sonication, $(NH_4)_2SO_4$, fractionation, and chromatography. The ZADH-I1 enzyme produced by Z. mobilis cell was also purified to compare to the enzyme produced by E. coli (pADS93). The purified enzyme from cell extract of E. coli (pADS93) was identified to be a tetramer being composed of four identical subunits having molecular weight of 40, 000 dalton like that of Z. mobilis. The pH optimum for the reaction oxidizing ethanol to acetaldehyde was 10.0 while the optimum for the reverse reaction was 7.5-8.5. The apparent $K_m$ values for ethanol and NAD + were $1.2 \times 10^{-1}M$and $5.1\times 10^{-5}M$, respectively. In addition, it was found that the $K_m$ value for acetaldehyde was very lower than that for ethanol.

  • PDF

Stability of Ethanolic Extract from Fermented Cirsium setidens Nakai by Bioconversion during Different Storing Conditions (저장조건에 따른 생물전환 발효고려엉겅퀴 주정추출물의 안정성 조사)

  • Lee, Jin-Ha;Moon, Seok-Yong;Cho, Bong-Yeon;Choi, Sun-Il;Jung, Tae-Dong;Choi, Seung-Hyun;Kim, Jong-Dai;Lee, Ok-Hwan
    • The Korean Journal of Food And Nutrition
    • /
    • v.30 no.2
    • /
    • pp.388-394
    • /
    • 2017
  • This study investigated the stability of ethanolic extract from fermented Cirsium setidens Nakai (FCSN) in order to develop functional materials during different storage conditions. We evaluated pectolinarin and pectolinarigenin contents, total phenol content and antioxidant activity (DPPH radical scavenging activity and FRAP assay) of ethanolic extract from FCSN obtained by bioconverison at various temperatures (4, 25 and $50^{\circ}C$) and pHs (4.0, 7.0 and 10.0). Our results show that the pectolinarin, pectolinarigenin, and total phenol contents in ethanolic extract from FCSN were decreased during the storage periods. Moreover, the DPPH radical scavenging activity did not significantly change at $4^{\circ}C$ and $25^{\circ}C$. Pectolinarin and pectolinarigenin contents, total phenol content and DPPH radical scavenging activity of ethanolic extract from FCSN at acidic pH (pH 4.0) and neutral pH (pH 7.0) were higher than those at the alkaline pH range. These results indicate that the optimum storage condition of the ethanolic extract from FCSN was $4^{\circ}C$ and pH 4.0~7.0 range.

${\gamma}$-Aminobutyric Acid Production and Glutamate Decarboxylase Activity of Lactobacillus sakei OPK2-59 Isolated from Kimchi (김치유래 Lactobacillus sakei OPK2-59의 ${\gamma}$-Aminobutyric Acid 생성 및 Glutamate Decarboxylase 활성)

  • Yu, Jin-Ju;Oh, Suk-Heung
    • Korean Journal of Microbiology
    • /
    • v.47 no.4
    • /
    • pp.316-322
    • /
    • 2011
  • Lactobacillus sakei OPK2-59 isolated from kimchi was found to have ${\gamma}$-aminobutyric acid (GABA) producing ability and glutamate decarboxylase (GAD) activity. When the Lactobacillus sakei OPK2-59 was cultured in MRS broth with 59.13 mM and 177.40 mM monosodium glutamate (MSG), the optimum temperature range and pH for growth were $25-37^{\circ}C$ and pH 6.5, respectively. GABA conversion rates in MRS broth with 59.13 mM and 177.40 mM MSG were 99.58% and 31.00%, respectively at $25^{\circ}C$ and 48 h of cultivation. By using the cell free extract of Lactobacillus sakei OPK2-59, MSG was converted to GABA and the conversion rate was 78.51% at $30^{\circ}C$, pH 5. Conversion of MSG to GABA was enhanced by adding salts such as $CaCl_2$, $FeCl_3$, $MgCl_2$. These data suggest that the ability of Lactobacillus sakei OPK2-59 to produce GABA results from the activity of GAD in the cells and GABA conversion by the cell extract containing GAD can be enhanced by $CaCl_2$, $FeCl_3$, $MgCl_2$.

Biodegradation of Phenanthrene by Transformant Trametes versicolor MrP1 (구름버섯의 형질전환체 Trametes versicolor MrP1에 의한 Phenanthrene의 생분해)

  • Choi, Yun-Seong;Choi, Hyoung-Tae;Song, Hong-Gyu
    • Korean Journal of Microbiology
    • /
    • v.43 no.4
    • /
    • pp.273-278
    • /
    • 2007
  • As a model compound of PAHs (polycyclic aromatic hydrocarbons) phenanthrene has been regarded as a toxic material, mutagen and carcinogen in various animals. Biodegradation conditions of phenanthrene such as pH, temperature, shaking speed, stabilizer and cofactor of degrading enzymes were investigated with Trametes versicolor and its transformant T. versicolor MrP1 in YMG medium, minimal medium and soil microcosm. T. versicolor MrP1 can overexpress mrp gene encoding Mn-repressed peroxidase that is involved in fungal degradation. Biodegradations of phenanthrene by T. versicolor and T. versicolor MrP1 were optimally performed in conditions of weak-acid (pH 6.0), $30^{\circ}C$, shaken culture and medium containing 5 mM veratryl alcohol or tryptophan. In these optimal conditions, biodegradation of phenanthrene by T. versicolor MrP1 is 31% higher than that of wild type strain in a minimal medium for 20 days. Biodegradation of phenanthrene by T. versicolor MrP1 was also higher than that of wild type in soil microcosm. T. versicolor MrP1 can be a excellent candidate for the bioremediation of PAHs contaminated environments.

Double-Stranded RNA-Dependent Protein Kinase Gene Expression in Tobacco Plant (연초식물체에서의 dsRNA 의존성 인산화 효소 유전자 발현)

  • 이청호;박희성
    • Korean Journal Plant Pathology
    • /
    • v.11 no.2
    • /
    • pp.173-178
    • /
    • 1995
  • 동물계에서 항바이러스와관련된 dsRNA 의존성 인산화 효소(PKR)의 유전자를 식물체에서 발현시킬 경우 PKR에 의한 단백질합성 및 식물바이러스의 증식조절 가능성에 대한 기초자료를 확보하기 위하여 사람에서 분리된 PKR cDNA를 Agrobacterium 방법에 의하여 연초식물체(Nicotiana tabacum cv. Xanthi-nc)로 형질전환시켰다. HindIII/PstI처리에 의해 얻어지는 약 1.8kb의 phPKR cDNA절편을 일련의 유전자 조작 방법을 통하여 식물발현벡터인 pBI121에 도입하여, p12168을 재조합하였다. 이를 A. tumefaciens LBA 4404에 형질전환시켜 연초식물체형질 전환에 이용하였다. 2mg/l BA와 0.5mg/l NAA가 포함되고 100$\mu\textrm{g}$/ml의 kanamycin이 첨가된 MS배지에서 shooting시킨 후 phytohormone이 첨가되지 않은 MS배지상에서 rooting을 시켜 형질전환 연초식물체를 얻었으며, 형질전환식물체는 정상식물체와 유사한 생육양상을 나타내었다. 형질전환식물체의 유전자도입은 hPKR cDNA의 전사부여는 RT-PCR 방법에 의하여 확인되었다.

  • PDF

Increased Expression of a Chemically Synthesized Human Lysozyme Gene in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 합성된 Human Lysozyme 유전자의 발현증대)

  • 김기운;최선욱;이승철;백현동;황용일
    • Microbiology and Biotechnology Letters
    • /
    • v.26 no.1
    • /
    • pp.34-39
    • /
    • 1998
  • We have already prepared a human lysozyme (HLY) structural gene from chemically synthesized 38 oligomers with high codon usage in Saccharomyces cerevisiae. For directing the synthesis and secretion of HLY in S. cerevisiae, two types of expression vectors, a YCp centromere-based vector, pHK101 and a YEp 2-$\mu\textrm{m}$ circle-based vector, pHK501 were constructed. With the resulting plasmids, we have confirmed that yeast transformant harboring pHK501 has more secreted HLY than pHK101-transformant by using a lysoplate and a turbidimetric assay. In flask cultivation, pHK501-transformant produced active HLY about 8 times (55 units/$m\ell$) higher than pHK101-transformant. From batch cultivation, the HLY productivity was obtained with 1.12 units/$m\ell$/h, corresponding to a 1.8-fold increase compared with flask fermentation. These results indicate that yeast transformant with pHK501 vector overexpressed and secreted HLY than that of YCp type vector.

  • PDF

Influence of Performance and Microbial Community by Internal pH Control on Anaerobic Digestion of Food Waste Leachate (음폐수 이용 혐기성 소화의 내부 pH 조절에 따른 바이오가스 전환율 비교 및 미생물 군집도 분석)

  • Yun, Yeo-Myeong;Cho, Si-Kyung;Jeong, Da-Young;Lee, Eun-Jin;Huh, Kwan-Yong;Shin, Dong-Hyuk;Lee, Chang-Kyu;Shin, Hang-Sik
    • Journal of Korean Society of Environmental Engineers
    • /
    • v.35 no.8
    • /
    • pp.571-578
    • /
    • 2013
  • In this study, the performance and microbial community of anaerobic digestion fed by food waste leachate at low organic loading rate were investigated with and without internal pH control. Experimental results show that similar biogas yield was achieved in both reactors regardless of increase in pH, the concentrations of free ammonia and volatile fatty acids in case of without internal pH controlled one. The results of a methanogenic community analysis by Polymerase Chain Reaction and Denaturing Gradient Gel Electrophoresis revealed that the apparent preponderance of Methanosarcina sp. could be one of reasons for the maintenance of reactor stability.

Catalytic Activity of Ga(Ⅲ)-, In(Ⅲ)- and Tl(Ⅲ)-porphyrin Complexes (Ga(Ⅲ), In(Ⅲ) 및 Tl(Ⅲ) 금속이온을 포함한 Metalloporphyrin 착물의 촉매적 특성)

  • Park, Yu Chul;Na, Hun Gil;Kim, Seong Su
    • Journal of the Korean Chemical Society
    • /
    • v.39 no.5
    • /
    • pp.364-370
    • /
    • 1995
  • The catalytic oxidations of several olefins in $CH_2Cl_2$ have been investigated using non-redox metalloporphyrin (M=Ga(III), In(III), Tl(III)) complexes as catalyst and sodium hypochlorite as terminal oxidant. Porphyrins were $(p-CH_3O)TPP,\;(p-CH_3)TPP,\;TPP,\;(p-F)TPP,\;(p-Cl)TPP\;and\;(F_20)TPP$ (TPP=tetraphenylporphyrin), and olefins were $(p-CH_3O)-,\;(p-CH_3)-,\;(p-H)-,\;(p-F)-,\;(p-Cl)-\;and\;(p-Br)styrene$styrene and cyclopentene and cyclohexene. The substrate conversion yield was discussed according to the substituent effects of metalloporphyrin and substrate, and the radius effect of non-redox metal ion. The conversion yield of substrate by changing the substituent of TPP increased in the order of $p-CH_3O$ < $p-CH_3$ < H < p-F < p-Cl, which was consistent with the sequence of $4{\sigma}$ values of TPP. But the substituent effect of substrate on the conversion yield decreased with increasing the ${\sigma}^+$ values on substrates in the order of p-CH3O > p-CH3 > H > p-Cl > p-Br. For the oxidation of several olefins, the complexes of In(III)- and Tl(III)-porphyrins turned out to be more active catalysts than Ga(III)-porphyrin.

  • PDF

Allergic risk assessment of genetically modified cucumber mosaic virus resistant pepper (유전자변형 바이러스 저항성 고추의 알레르기 안전성)

  • Son, Dae-Yeul
    • Food Science and Preservation
    • /
    • v.22 no.6
    • /
    • pp.901-907
    • /
    • 2015
  • Genetically modified (GM) pepper H15 containing the gene for cucumber mosaic virus (CMV) coat protein (CP) and its control line non-GM pepper P2377 were investigated for their allergic risk. Amino acid sequence of the inserted gene product CMV-CP was compared with those of known allergens. No known allergen had greater than 35% amino acid sequence homology over an 80 amino acid window or more than 8 consecutive identical amino acids. Protein patterns of GM and non-GM pepper extracts were evaluated by SDS-PAGE, which showed similar distribution of protein bands for both GM and non-GM pepper. Antigen-antibody reactions were compared between GM and its non-transgenic parental control. ELISA and immunoblot analysis of sera from allergic patients showed some IgE reactivity; however, no differences were observed between GM pepper H15 and P2377. We therefore conclude that CMV-CP is less likely to be an allergen; the protein composition and allergenicity of the GM pepper H15 is not different from that of P2377 and safe as a commercial host.

Stable Transformation via Callus Formation and Rhizogenesis from the Cultures of Hypocotyl Explant of Chinese Cabbage (배추의 배축절편으로부터 캘러스와 뿌리 발생을 통한 안정적 형질전환)

  • Cho, Mi-Ae;Kim, Choon-Ae;Min, Sung-Ran;Ko, Suck-Min;Liu, Jang-Ryol;Choi, Pil-Son
    • Journal of Plant Biotechnology
    • /
    • v.34 no.2
    • /
    • pp.139-144
    • /
    • 2007
  • Hypocotyl explants of Chinese cabbage (cvs. "Jeong Sang") produced transgenic calli on callus induction medium (MS salt, B5 vitamin, 5 mg/L acetosyringone, 1 mg/L 2,4-D, 3% sucrose, 400 mg/L cefotaxime, 100 mg/L paromomycin, pH 5.8) after cocultivation with strains of Agrobacterium tumefaciens (EHA101, LBA4404, GV3101) harboring the pPTN290 containing paromomycin-resistance gene as a selectable marker, and then they transferred to root induction medium (1/2MS salt, MS vitamins, 2% sucrose, 100 mg/L paromomycin, 100 mg/L cefotaxime, pH 5.8) and shoot induction medium (MS salt, B5 vitamin, 4 mg/L $AgNO_3$, 4 mg/L 6-benzyladenine, 3 mg/L alpha-naphthaleneacetic acid, 100 mg/L paromomycin, 100 mg/L cefotaxime, 3% sucrose, pH 5.8) in order. There was a significant difference in the frequency of transgenic calli depending on Agrobacterium strains. In particular, the highest frequency (6.1%) of transgenic calli was obtained from the hypocotyls cocultivated with EHA101 strains. Also, the frequency (%) of transgenic root and plants from each transgenic callus clone were obtained with 60.7% and 38.2% in EHA101, with 8.3% and 0% in LBA4404, with 20.5% and 85.7% in GV3101 strains, respectively. They were grown to maturity in a greenhouse and normally produced $T_2$ seeds. GUS histochemical assay for progeny ($T_2$) revealed that the transgenes was expressed in the plant genome, and progeny analysis from 7 independent transgenic events demonstrated that the transformants transmitted the transgene as a single or multiple functional locus.