• Title/Summary/Keyword: C&DH

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Analysis of the functional domains of CFTase gene cloned from Xanthomonas oryzae #5 using recombinant deletion mutant

  • Kim, Byeong-U;Yu, Dong-Ju;Ryu, Hye-Gyeong;Park, Ju-Hui
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.727-730
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    • 2001
  • Various recombinant deletion mutants were constructed from cycloinulo - oligosaccharide fructanotransferase(CFTase) gene of Xanthomonas oryzae #5 . The mutants were expressed in Escherichia coli DH5${\alpha}$. We were able to obtain three recombinant proteins were purified, and examine their CFTase and hydrolyzing activity. N-terminal deletion mutant had both CFTase activity and hydrolyzing activity. however, in C-terminal and N,C-terminal deletion mutant disappeared CFTase activity, but hydrolyzing activity remained. From there results, it seems that the C-terminal region(amino acid $1173{\sim}1333$) is important for cyclization.

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Hydrolysis of Isolate Soybean Protein Using Subcritical Water (아임계수를 이용한 분리대두단백질의 가수분해)

  • Hwang, Yun Hee;Cho, Hyung-Yong;Kim, Ko-Rae;Lee, Seok Hoon;Choi, Mi-Jung;Shin, Jung-Kue
    • Korean Journal of Food Science and Technology
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    • v.47 no.6
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    • pp.772-778
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    • 2015
  • Hydrolysis of isolate soybean protein (ISP) using subcritical water (SCW) was conducted to study the feasibility for producing protein hydrolyzate. SCW hydrolysis of SPI suspension (5-15%) was conducted in an electrically heated batch reactor (2 L). The effects of temperature (230 to $270^{\circ}C$) and holding time (10 to 50 min) on the degree of hydrolysis (DH) and the production of amino acids were studied by surface response method. The DH was determined by derivatizing the hydrolyzates with ortho-phthalaldehyde (OPA) solution. It was confirmed that reaction temperature and holding time affected the hydrothermolysis of soybean protein. However, the holding time was less effective on amino acid yield when the temperature was higher than $230^{\circ}C$. In order to achieve optimal yields of amino acids exceeding 43%, the temperature should be within the range between 256 and $268^{\circ}C$ with holding time from 29 to 41 min, respectively. A maximum estimated amino acid yield of 43.5% was obtained at $268^{\circ}C$ for 35 min.

Cloning and Characterization of Cellulase Gene (cel5C) from Cow Rumen Metagenomic Library (소 반추위 메타게놈에서 새로운 섬유소분해효소 유전자(cel5C) 클로닝 및 유전산물의 특성)

  • Kim, Min-Keun;Barman, Dhirendra Nath;Kang, Tae-Ho;Kim, Jung-Ho;Kim, Hoon;Yun, Han-Dae
    • Journal of Life Science
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    • v.22 no.4
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    • pp.437-446
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    • 2012
  • A metagenomic library of cow rumen in the pCC1FOS phage vector was screened in $E.$ $coli$ EPI300 for cellulase activity on carboxymethyl cellulose agar plates. One clone was partially digested with $Sau$3AI, ligated into the $Bam$HI site of the pBluescript II SK+ vector, and transformed into $E.$ $coli$ $DH5{\alpha}$. We obtained a 1.5 kb insert DNA, designated $cel$5C, which hydrolyzes carboxymethyl cellulose. The cel5C gene has an open reading frame (ORF) of 1,125 bp encoding 374 amino acids. It belongs to the glycosyl hydrolase family 5 with the conserved domain LIMEGFNEIN. The molecular mass of the Cel5C protein induced from $E.$ $coli$ $DH5{\alpha}$, as analyzed by CMC SDS-PAGE, appeared to be approximately 42 kDa. The enzyme showed optimum cellulase activity at pH 4.0, and $50^{\circ}C$. We examined whether the $cel$5C gene comes from the 49 identified cow rumen bacteria using PCR. No PCR bands were identified, suggesting that the $cel$5C gene came from the unidentified cow rumen bacteria.

Initial Operating Power Control of Redox Flow Battery in Offgrid 10kW ESS (독립형 10kW ESS의 레독스 플로우 배터리 초기 구동 전원 제어)

  • Choi, JM;Song, YH;Choi, ES;Eun, JM;Lee, YJ;Han, DH;Cho, YH;Choe, GH
    • Proceedings of the KIPE Conference
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    • 2013.07a
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    • pp.390-391
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    • 2013
  • This paper proposes advanced RFB PCS for islanded environment. To accommodate islanded system, power conditioning needs voltage control authority changing. RFB initial power generating method is proposed for the Islanded PCS. DC-link voltage control authority is changed to PV converter to bidirectional converter by proposed control logic. The control performance has been verified with hardware experiments.

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Reduction of the Antigenicity of Whey Protein by Enzymatic Hydrolysis (효소가수분해에 의한 유청단백질의 항원성 저하)

  • Ha, Woel-Kyu;Juhn, Suk-Lak;Kim, Jung-Wan;Lee, Soo-Won;Lee, Jae-Young;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.26 no.1
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    • pp.74-80
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    • 1994
  • As a preliminary study about the reduction of the antigenicity of whey protein isolate(WPI) by treatment of chymotrypsin, trypsin, pancreatin, and protease from Aspergillus oryzae, the properties and antigenicities of whey protein hydrolysates(WPH) were investigated. When degrees of hydrolysis (DH) were measured by use of trinitrobenzensulfonic acid(TNBS), the DH of the WPH treated by pancreatin or protease from Aspergillus oryzae$(5.05{\sim}11.47)$ were much higher than those of the tryptic or chymotryptic WPH$(15.67{\sim}20.20)$. And the pretreatments of heat$(75^{\circ}C)$, 20 min and/or pepsin resulted in higher DH of WPH, generally. When the molecular distributions of the WPH were determined by high performance size exclusion chromatography(HPSEC), the ratios of polypeptides with molecular weight more than 10kDa ranged from 12% to 36%, and the average molecular weights and the average peptide lengths of the WPH were $4,252{\sim}9,132$ dalton and $38{\sim}83$ amino acids, respectively. And there was no bitter taste in all of the WPH. Results of SDS-PAGE showed that most of intact native proteins were eliminated by the enzymatic hydrolysis but there were a few bands of peptides larger than 14.2 kDa in some WPH. When antigenicity was assayed by competitive inhibition enzyme-linked immunosorbent assay(cELISA), monovalent antigenicity of WPH to rabbit anti-WPI antiserum were lowered to $10^{-1.7}-10^{-4.9}$ times and less by the enzymatic hydrolysis. And the pretreatments of heat and pepsin resulted in the lowest antigenicicy within a group of enzymatic hydrolysis, especially in case of the pancreatic hydrolysate(PDP) whose antigenicity was found almost to be removed.

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Genetic Mapping of QTLs that Control Grain Characteristics in Rice (Oryza sativa L.) (벼의 낱알 특성에 관여하는 양적형질유전자좌 분석)

  • Wacera, Home Regina;Safitri, Fika Ayu;Lee, Hyun-Suk;Yun, Byung-Wook;Kim, Kyung-Min
    • Journal of Life Science
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    • v.25 no.8
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    • pp.925-931
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    • 2015
  • We performed a molecular marker-based analysis of quantitative trait loci (QTLs) for traits that determine the quality of the appearance of grains, using 120 doubled-haploid (DH) lines developed by another culture from the F1 cross between ‘Cheongcheong’ (Oryza sativa L. ssp. Indica) and ‘Nagdong’ (Oryza sativa L. ssp. Japonica). The traits studied included length, width, and thickness of the grains, as well as length-to-width ratio and 1,000-grain weight. The objective of this study was to determine the genetic control of these traits in order to formulate a strategy for improving the appearance of this hybrid. Within the DH population, five traits exhibited wide variation, with mean values occurring within the range of the two parents. Three QTLs were identified for grain length on chromosomes 2, 5, and 7. Three QTLs were mapped for grain width on chromosome 2: qGW2-1, qGW2-2, and qGW2-3. Six chromosomes were identified for the grain length-to-width ratio; four of these were on chromosome 2, whereas the other two were on chromosomes 7 and 12. One QTL influencing 1,000-grain weight was identified and located on chromosome 8. The results presented in the present study should facilitate rice-breeding, especially for improved hybrid-rice quality.

Characteristics of Heavy Metal Resistant Plasmid in Enterobacter cloaceae K41 (Enterobacter cloaceae K41 plasmid의 중금속 저항성)

  • Kim Young-Hee;Lee Sang-Jun;Jeong Yong-Kee;Chung Kyung-Tae
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.566-571
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    • 2005
  • A natural habitat bacterium, Enterobacter cloaceae K41 was isolated from fresh water plant root and identified. This strain was used to investigate heavy metal resistance. The optimal growth conditions of the bacterium were LB medium containing$1\%$ yeast extract, $1\%$ lactose, $1\%$ NaCl, pH 7.0, at $37^{\circ}C$, and for 24 hours on a shaker. The minimal inhibitory concentration (MIC) of heavy metals against E. cloaceae KCTC2519 and E. cloaceae K41 was compared. The MIC of E. cloaceae K41 was 150 ppm in Cu, 50 ppm in Cd whereas that of the standard strain was 50 ppm in Cu but no growth was observed either Cd or two mixed heavy metal solution. The presence of plasmid was cleared from the isolated strain whereas no possession from the standard strain. The plasmid from E. cloaceae K41 was transformed into E. coli $DH5{\alpha}$. The MIC of transformed strain increased resistance 7 times in Cu and 6 times in Cd by insertion of this plasmid. The metal adsorption of the transformant was increased 1.3 times in Cu and 1.5 times in Cd indicating the plasmid was responsible for heavy metal resistance.

Expression of Human Immunodeficiency Virus Type 1 Gag Protein in Escherichia coli

  • Park, Weon-Sang
    • Journal of Life Science
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    • v.9 no.5
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    • pp.556-563
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    • 1999
  • Presence of antibody to the capsid protein p24 is the main diagnostic criterion, since this reflects reliable antibody response to HIV infection. However, it takes about 6-8 weeks for antibody production after infection and people who are infected but antibodies are not produced yet are classified as seronegative. Therefore, there is a strong need for an improved diagnostic method for better health security. As a first step for developing such an improved diagnostic system, gag protein of human immunodificiency virus type 1 was expressed in E. coli DH5$\alpha$. The gag fragment of HIV-1 (including a portion of p17 and whole p24) was amplified by polymerase chain reaction (PCR) and BamH I/EcoR I sites were created during PCR. The amplified DNA fragment was cleaved with BamH I/EcoR I and was subcloned into the GEX-2T vector which had been digested with BamHI/EcoRI, resulting gene fusion with gst gene of pGEX-2T. The recombinant DNA was transferred into E. coli DH5$\alpha$. The transformed bacteria were grown at 37$^{\circ}C$ for 3h and protein expression was induced with 0.1mM IPTG at $25^{\circ}C$ for 3h. Recombinant gag protein or GST-gag fusion protein was purified with glutathione-sepharose 4B bead and migrated as a single band when analyzed by 10% polyacrylamide gel. These proteins were confirmed by immunoblotting with anti-GST goat sera or Korean AIDS patients sera. The results of this study establish the expression and single step pulification of HIV-1 gag protein which can specifically bind with Korean AIDS patients sera.

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An Introduction to the Ground Water Model Test (지하수 model에 관한 모형시험방법)

  • 김주욱
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.9 no.2
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    • pp.1301-1305
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    • 1967
  • Ground water flow can be studied with model test. Model test of ground water works are necessary for economic and safe design of the works. Also influence of the ground water flow to the durability and safety of hydraulic structures can be studied with this model. a. Sand model ; Water flow through porous media is the principle of sand model. Darcy's formula is the basic equation, $q=k{\frac{dh}{ds}}^{\circ}. The effect of the ground water flow on the grain system itself is represented with this model only. b. Hele-Shaw model ; In this model use is made of the viscous flow analogy. Viscous fluid such as glycerine flowing through two parallel plates depends on Poiseuille law, $q=-c{\frac{dh}{ds}}$. The analogue can be used vertically and horizontally. c. Heat model ; This is based on the analogy of the Fourier's law for heat conduction and Darcy's law for ground water flow. Especially unsteady problem can be studied with this model. A difficulty of the construction of this model is the isolation, which has to prevent losses of the heat. d. Electirc model ; Ohm's law for electric current is analogous to Darcy's law. Resistance material such as metal foil, graphite block, water with salt added, gelatine with salt added, ete. is connected to electric sources and resistor, and equi-voltage line is detected with galvanometer, $N_aCl$, $CuSo_4$, etc. are used as salt in the model. e. Membrane model ; This model is based on the facts that the deflection of a thin membrane obeys Laplace's equation if there is no load in the direction perpendicular to the membrane, and if the dellection is small.

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The CT values Comparisons for Antibiotic Resistant Bacteria and Resistant Genes by Chlorination (항생제 내성균 및 유전자제거를 위한 염소 CT 값 비교)

  • Oh, Jun-Sik;Kim, Sungpyo
    • Journal of Wetlands Research
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    • v.16 no.2
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    • pp.269-274
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    • 2014
  • The purpose of this study is to compare CT (disinfectant concentration * time) values in removing the antibiotic resistance bacteria, antibiotic resistance gene and transfer of antibiotic resistance genes. Different concentration of chlorine(C) and contact time(T) according to the removal of antibiotic resistance was calculated for each. As a result, for the 90% removal of antibiotic resistant bacteria, around 176~353 mg min/L CT values are needed. For the removal of the antibiotic resistance gene, 195~372 mg min/L CT values are required. For the 90% reduction of antibiotic resistance gene transfer by chlorine disinfection, 187~489 mg min/L CT values are needed. Based on our results, higher CT value was required for removing antibiotic resistant genes rather than antibiotic resistance bacteria.