• Title/Summary/Keyword: Synthesis Conditions

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Molecular Cloning of Glycoside Hydrolase Family 74 Genes and Analysis of Transcript Products from the Basidiomycete Phanerochaete chrysosporium (담자균 Phanerochaete chrysosporium으로부터 유래한 Glycoside Hydrolase Family 74 유전자 클로닝과 전사산물 분석)

  • Lee, Jae-Won;Samejima, Masahiro;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.3
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    • pp.56-63
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    • 2006
  • In order to evaluate the mechanism of cellulose hydrolysis, the complementary DNA encoding Glycoside Hydrolase Family (GHF)74 was cloned from Phanerochaete chrysosporium. Depending on the presence of Cellulose Binding Module (CBM), it can be classified as GHF74A or GHF74B. The GHF74A gene from P. chrysosporium (PcGHF74A) consists of 2163 bp encoding a protein of 721 amino acid residues. The PcGHF74A showed homology of 70~77% compared with the GHF74 from other filamentous fungi. The PcGHF74B, which contains CBM and is a member of family 1, was transcribed to various transcripts depending on the nature of carbon sources and their concentration. To study the possible presence of splice variants in GHF74B transcripts in P. chrysospoium, we carried out RT-PCR analysis using primers that designed based on the annotation data and sequenced data. Our result indicated that PcGHF74B was transcribed to several splicing variants in various culture conditions. Especially in the culture of 2% cellulose, three transcript products were observed. First transcript was presumed to be a full length ORF that contained 11th intron with stop codon at position 2562 bp. The second one consisted of 12 exons and 11 introns with stop codon at position 1187 bp with 7th exon. The shortest transcript consisted of 10 exons and 9 introns with stop codon at 910 bp in the 7th exon. These premature stop codon might prevent the synthesis of fully active GHF74 or contribute for the production of protein with distinct function depending on the ambient carbon sources.

${\beta}$-1,4-Xylosidase Activity of Leuconostoc Lactic Acid Bacteria Isolated from Kimchi (김치에서 분리된 Leuconostoc 속 젖산균의 ${\beta}$-1,4-xylosidase 효소생산 특성)

  • Jang, Mi-Hee;Kim, Myoung-Dong
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.169-175
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    • 2011
  • The ${\beta}$-xylosidase (EC 3.2.1.37) production capabilities of lactic acid bacteria in the genus Leuconostoc, isolated from a variety of kimchi (fermented vegetables), were examined. The intracellular levels of ${\beta}$-xylosidase were similar to the extracellular levels, when most Leuconostoc lactic acid bacteria were grown in a medium containing xylose as the carbon source. Intracellular ${\beta}$-xylosidase with a maximum activity of $1.2{\pm}0.1units/mL$ (mean${\pm}$standard error) was obtained from Leuconostoc lactis KCTC 13344, which was isolated from fermented Chinese cabbage. The optimum reaction conditions for Leu. lactis KCTC 13344 ${\beta}$-xylosidase activity were pH 6.0 and $30^{\circ}C$, and the addition of most divalent cations, except zinc, to the reaction mixture resulted in a slight increase in enzyme activity. Compared with a media containing other carbon sources, the ${\beta}$-xylosidase activity was 5 times higher when Leu. lactis KCTC 13344 was grown in a medium containing xylose as carbon source. Zymographic analysis indicated that the synthesis of Leu. lactis KCTC 13344 ${\beta}$-xylosidase (approximate size, 64 kDa) is induced by xylose. A maximum intracellular ${\beta}$-xylosidase activity of $7.1{\pm}0.3units/mL$ was obtained in a batch cultivation in an MRS medium containing 30 g/L xylose.

Synthesis of Functional Lipid from Glyceryl Monooleate and Conjugated Linoleic Acid by Enzymatic Reaction (Glyceryl Monooleate와 Conjugated Linoleic Acid로부터 효소적 반응을 이용한 기능성 유지 합성)

  • Jeon, Mi-Sun;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.8
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    • pp.1062-1068
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    • 2009
  • Diacylglycerol (DAG) were synthesized by enzymatic esterification of glyceryl monooleate (GMO) and conjugated linoleic acid (CLA) in a shaking water bath. The reaction was catalyzed by Lipozyme TLIM (immobilized lipase from Thermomyces lanuginosa). Effects of reaction time, molar ratio, enzyme road and molecular sieves were studied. Results of normal-phase high performance liquid chromatography (NP-HPLC) analysis were performed. At 1:1, 2:1 and 3:1 (GMO : CLA) molar ratio and Lipozyme TLIM of 20% amount, DAG were produced in 42.6, 54.4 and 54.6 area% in 1 hr, respectively. When different Lipozyme TLIM amounts (2, 5, 10, 20%) were used with 2:1 (GMO : CLA) molar ratio, DAG were produced 21.4 (24 hr), 51.7 (12 hr), 56.2 (6 hr) and 54.4 (1 hr) area%, respectively. The reaction in the absence of molecular sieves increased DAG contents. The maximum DAG concentration conditions were obtained with molar ratio of 2:1 (GMO : CLA), lipase concentration of 10% (of substrate), 10% molecular sieves and reaction time of 6 hours at 55$^{\circ}C$. Under this reaction condition, produced DAG-rich oil was composed of 69 area% DAG, 7.9 area% TAG, 2 area% FFA, and 21.1 area% MAG.

Development and Application of Enzyme Immunoassay for Endosulfan Residue Analysis (Endosulfan 과 그 분해산물의 Enzyme Immunoassay에 의한 분석법의 개발과 응용)

  • Suh, Yong-Tack;Shim, Jae-Han;Lee, Kang-Bong
    • Korean Journal of Environmental Agriculture
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    • v.11 no.1
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    • pp.59-66
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    • 1992
  • An enzyme immunoassay(EIA) was developed for the analysis of insecticide endosulfan and its degradation products. The sensitivity and specificity of the antibody produced were examined. Optimal conditions in the ELISA system for residue analysis were also discussed. A mixed suspension of endosulfan-hemocyanin conjugate(ES-KLH) 1.1 mg/ml and Freund's adjuvant was injected subcutaneously to white rabbits and then collected antisera were tested for titers by indirect ELISA(1/24,000). Because of difficulties in the synthesis of endosulfan peroxidase conjugate, amine derivative of endosulfan-diol was synthesized and it showed 40% of conjugate yields(2mg/ml of conjugate). the highest sensitivity obtained enzyme-conjugate was a concentration of 200ng/ml. The detection limit of endosulfan in ELISA system was 5 ppb on the standard curve. In application of ELISA for residue analysis, the recoveries were really 100% both in the spiked soil and apple sample regardless of endosulfan concentration treated. On the other hand, chlorinated hydrocarbons of similar structure with endosulfan showed low cross-reactivity$(2.2%{\sim}29.2%).$

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Properties of Antimicrobial Substances Produced by Bacillus amyloliquefaciens CJW15 and Bacillus amyloliquefaciens SSD8 (Bacillus amyloliquefaciens CJW15와 SSD8이 만드는 항균물질들의 특성)

  • Liu, Xiaoming;Shim, Jae Min;Yao, Zhuang;Lee, Jae Yong;Lee, Kang Wook;Kim, Hyun-Jin;Ham, Kyung-Sik;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.9-18
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    • 2016
  • Two Bacillus strains, CJW15 and SSD8, with strong antibacterial activities were isolated from cheonggukjang. Both were identified as B. amyloliquefaciens strains after gene sequencing of rRNA and recA. CJW15 strongly inhibited the growth of B. cereus (ATCC14579), Listeria monocytogenes (ATCC19111), and Lactococcus lactis (ATCC11454). In comparison, SSD8 inhibited the growth of B. cereus (ATCC14579) and Enterococcus faecium (ATCC19953). The antibacterial activities of the two strains were not affected when exposed to a temperature of $100^{\circ}C$ for 15 min and were quite stable in acidic (pH 3) and alkaline (pH 12) pH conditions. Enzymatic treatments (trypsin, pepsin, proteinase K, and protease) had no effect on the activity of CJW15, but reduced the activity of SSD8 by half. Both isolates possess genes involved in the synthesis of lipopeptides (e.g. surfactin, fengycin, iturin, and iturin A), and genes encoding subtilin, a bacteriocin. Moreover, both isolates have fibrinolytic activities as well.

Synthesis of Diglyceride Containing Caprylic acid by Immobilized Lipase Catalyzed Esterification of Monoglyceride in a Solvent Free System (모노글리세리드와 카프릴산으로부터 고정화 리파제를 사용한 디글리세리드 생산)

  • Lee, Jang-Woon;Kang, Sung-Tae
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.365-370
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    • 2009
  • For the production of diglyceride (DG) containing medium chain fatty acid, which could be utilized as a substrate to structured lipid production, monoglyceride (MG) and caprylic acid were reacted in the presence of lipase. The reaction system was well mixed homogeneously without using any organic solvent. Among the lipases investigated, Lipozyme RM IM and Novozym 435 were selected on the basis of equilibrium DG yields from the medium chain fatty acid and MG. And reaction conditions such as addition of molecular sieve, water content of immobilized lipase, reaction temperature, and mole ratio of MG/caprylic acid are optimized to increase DG production by using Lipozyme RM IM. DG content of reaction mixture showed 8% increase by adding molecular sieve to reaction mixture. Removal of water from the immobilized lipase could affect seriously equilibrium content of DG. More than 2.8%(w/w) removal of water from the support could make 44% of DG. Optimum temperature was found to $60^{\circ}C$. Temperature shift from $60^{\circ}C$ to $25^{\circ}C$ resulted in increase of free fatty acid (FFA) content. The equilibrium DG yield was not seriously affected by on MG/caprylic acid molar ratio. However, at the stoichiometric ratio of 1:1 the highest DG yield was obtained. Increasing MG/caprylic acid ratio from 0.3 to 1.8 decreased FFA content from 34% to 13%, while MG content increased from 27% to 50%.

Seagrasses in Northern Chinese Seas: Historical Declines and Case Study of the Status (중국 북부 연안의 잘피: 역사적 감소추세 및 현황에 대한 사례)

  • Zhang, Xuelei;Li, Yan;Liu, Ping;Sun, Ping;Wang, Xiao;Fan, Shiliang;Xu, Qinzeng
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.13 no.4
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    • pp.305-312
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    • 2010
  • Seagrass beds are a type of coastal wetland with many ecosystem services and precious economic values. Seagrass meadows used to be widespread along the coasts in northern Chinese seas, yet they have long been overlooked and lack devoted study on their history and status. This paper firstly reveals, by synthesis of information on composition of seagrass species and their distribution, that the seagrasses in this region have experienced considerable declines, both in terms of distribution and biomass, from the earliest record to present days. Then, a case study at the seagrass bed of Chudao is described to show the status of representative seagrass meadows. The results indicate that the environmental condition is good, seagrasses are in recovery, the planktoners are healthy and rich fishery resources and the mammal finless porpoise are associated with the seagrass bed. The cause(s) of historical seagrass decline and current conditions are also discussed, and future recommendations on seagrass protection and mapping are suggested.

Isolation and Characterization of Bacterial Cellulose-Producing Bacteria for Silver Nanoparticle Synthesis (은 나노입자 합성을 위한 Bacterial Cellulose 생산 세균의 분리 및 특성)

  • Yoo, Ji-Yeon;Jang, Eun-Young;Son, Yong-Jun;Park, Soo-Yeun;Son, Hong-Joo
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.120-126
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    • 2018
  • As a basic study for environment-friendly production of bacterial cellulose (BC) dressing with antimicrobial activity, we isolated and identified acetic acid bacteria which are resistant to silver ions and can biosynthesize silver nanoparticles. Furthermore, conditions of BC production by selected strain were also investigated. Strain G7 isolated from decayed grape skin was able to grow in the presence of 0.1 mM $AgNO_3$ which was identified as Acetobacter intermedius based on 16S rRNA gene analysis. BC production was the highest in a medium containing 2% glucose as a carbon source, 2% yeast extract as a nitrogen source, and 0.115% acetic acid as a cosubstrate. Structural properties of BC produced in optimal medium were studied using Fourier-transform infrared spectroscopy and X-ray diffractometer, and it was found that BC produced was cellulose type I that was the same as a typical native cellulose. When strain G7 was cultured in an optimal medium containing 0.1 mM $AgNO_3$, the color of the culture broth turned into reddish brown, indicating that silver nanoparticles were formed. As a result of UV-Vis spectral analysis of the culture, it was found that a unique absorption spectrum of silver nanoparticles at 425 nm was also observed. Scanning electron microscopic observations showed that silver nanoparticles were formed on the surface and pores of BC membrane.

Importance of Selecting The characterized Housekeeping Genes as Reference Genes in Various Species (다양한 종에서 하우스키핑 유전자 선택의 중요성)

  • Chai, Han-Ha;Noh, Yun Jeong;Roh, Hee-Jong;Lim, Dajeong
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.8
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    • pp.417-428
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    • 2020
  • Housekeeping genes are expressed in cells of all organisms and perform basic cellular functions such as energy generation, substance synthesis, cell death, and cell defense. Accordingly, the expression levels of housekeeping genes are relatively constant, and thus they are used as reference genes in gene expression studies, such as protein expression and mRNA expression analysis of target genes. However, the levels of expression of these genes may be different among various tissues or cells and may change under certain circumstances. Therefore, it is important to select the best reference gene for specific gene expression research by exploring the stability of housekeeping gene expression. This review summarizes housekeeping genes found in humans, chickens, pigs, and rats in the literature and estimates expression stability using geNorm, NormFinder, and BestKeeper software. The most suitable reference housekeeping gene can selected based on expression stability according to the experimental conditions of the gene expression study and can thus be applied to data normalization.

Initial Risk Assessment of Acetanilide with Respect to Ecological Integrity (아세트아닐리드의 초기 환경위해성 평가)

  • Lee, Su-Rae;Park, Seon-Ju;Lee, Mi-Kyung;Nam, U-Kyung;Chung, Sun-Hwa;Seog, Geum-Su;Park, Kwang-Sik;Kim, Kyun;Kim, Yong-Hwa
    • Environmental Analysis Health and Toxicology
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    • v.15 no.1_2
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    • pp.19-29
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    • 2000
  • Acetanilide may be released into the environment through air and wastewater from its production and use sites as an intermediate in the synthesis of pharmaceuticals and dyes. Acetanilide is biodegraded rapidly under aerobic conditions and decomposed by indirect photolysis in the presence of OH radicals. An estimated bioconcentration factor of 4.5 suggests that bioaccumulation in aquatic organisms is low. Ecotoxicological data on acetanilide exist on acute toxicity to fishes of 4 species only. According to the EUSES system, the lowest PNEC (Predicted no effect concentration) in fishes is 0.01 mg/1 and PEC (Predicted environmental concentration) for surface water on a regional scale is 9.1$\times$10$\^$-5/mg/l as the worst case. RCR (Risk characterization ratio) of acetanilide for surface water on a regional scale was estimated as 9.1$\times$10-3, which is safe enough for fishes, RCR on a local basis slightly exceeds the value 1 in water and sediment; that is, 1.3 and 1.6, respectively, which suggests the existence of ecotoxicological risk at the vicinity of the manufacturing site. For the refinement of environmental risk assessment on acetanilide, more data should be collected regarding prolonged fish toxicity, acute toxicity toward daphnia and algae. It is, therefore, recommended that acetanilide should be a candidate for further work to supplement the lacking data until it is proved to be safe in the ecotoxicological aspects.

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