• Title/Summary/Keyword: hydrolysis activity

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Production and Characterization of Cyclodextrin Glucanotransferase fronm Bacillus sp. JK-43 Isolated from Kimchi (김치 분리균인 Bacillus sp. JK-43이 생산하는 Cyclodextrin Glucanotransferase의 생산 및 특성)

  • Jun, Hong-Ki;Bae, Kyung-Mi;Kim, Young-Hee;Baik, Hyung-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.1
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    • pp.41-48
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    • 2000
  • A bacterial strain, designated as JK-43, producing extracellular cyclodextrin glucanotransferase (CGTase)[EC 2.4.1.19] was isolated from kimchi. The CGTase from isolated strain JK-43 showed the transglucosylation activity from soluble starch to L-ascorbic acid(AA) compared to those obtained from other strains. A main product formed by this reaction was identified as $2-O-{\alpha}-glucopyranosyl$ L-ascorbic acid(AA-2G) by testing its susceptibility to ${\alpha}-glucosidase$ hydrolysis, the HPLC profiles, and through the elementary analysis. the ${\beta}-CD,\;{\gamma}-CD$, potato starch and corn starch were identified to be suitable glucosyl donor for transglucosylation reaction on AA by CGTase. Acceptor specificity on AA-2G production was examined by use of AA, Iso-AA and AA-2P. Transglucosylation was observed toward AA-2P as well as AA and Iso-AA. The microorganism isolated from kimchi was identified as a strain of Bacillus sp. JK-43 based on the morphological, cultural, biochemical characteristics and partial 16SrDNA sequence analysis. The maximal CGTase production was observed in a medium containing 1.0% soluble starch, 1.0% yeast extract, 1.0% $Na_2CO_3\;0.1%\;K_2HPO_4,\;and\;0.02%\;MgSO_4{\cdot}7H_2O$ with initial pH 7.0. The strain was cultured at $37^{\circ}C$ for 26 hrs with reciprocal shaking.

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Analysis of an Immobilized β-Galactosidase Reactor with Competitive Product Inhibition Kinetics (경쟁적 저해를 갖는 고정화 β-galactosidase 반응기의 해석)

  • Kang, Byung Chul
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1471-1476
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    • 2013
  • The present study deals with the immobilization of Kluyveromyces lactis ${\beta}$-galactosidase on a weak ionic exchange resin (Duolite A568) as polymer support. ${\beta}$-Galactosidase was immobilized using the adsorption method. A kinetic study of the immobilized enzyme was performed in a packed-bed reactor. The adsorption of the enzyme followed a typical Freundlich adsorption isotherm. The adsorption parameters of k and n were 14.6 and 1.74, respectively. The initial rates method was used to characterize the kinetic parameters of the free and immobilized enzymes. The Michaelis-Menten constant ($K_m$) for the immobilized enzyme (120 mM) was higher than it was for the free enzyme (79 mM). The effect of competitive inhibition kinetics was studied by changing the concentration of galactose in a recycling packed-bed reactor. The kinetic model with competitive inhibition by galactose was best fitted to the experimental results with $V_m$, $K_m$, and $K_I$ values of 46.3 $mmolmin^{-1}mg^{-1}$, 120 mM, and 24.4 mM, respectively. In a continuous packed-bed reactor, increasing the flow rate of the lactose solution decreased the conversion efficiency of lactose at different input lactose concentrations. Continuous operation of 11 days was conducted to investigate the stability of a long-term operation. The retained activity of the immobilized enzymes was 63% and the half-life of the immobilized enzyme was found to be 15 days.

Synthesis of Cyclododecatriene from 1,3-Butadiene by Trimerization over Amine-Titanium Complex Catalyst (아민-티타늄착체 촉매상에서 1,3-부타디엔의 삼량화반응에 의한 싸이클로도데카트리엔의 합성)

  • Park, Da Min;Kim, Gye-Ryeong;Lee, Ju Hyun;Cho, Deuk Hee;Kim, Geon-Joong
    • Korean Chemical Engineering Research
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    • v.51 no.3
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    • pp.394-402
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    • 2013
  • The new complex catalysts were synthesized by the reaction of titanium compounds (titanium chloride or titanium butoxide) and diamines in this work, and they showed very high catalytic activities for the cyclododecatriene (CDT) synthesis from 1,3-butadiene through trimerization. CDT synthetic reaction was performed in an autoclave reactor, and the effects of reaction temperature, type of catalyst, catalyst amount added into the system, the mole ratio of Al/Ti and immobilization method were investigated on the yield of product CDT. The titanium complex catalyst combined to diamine with 1:1 ratio showed high selectivity to CDT more than 90%. The ratio of TTT-CDT/TTC-CDT isomers in the product revealed as different values, depending on the type of diamine combined to titanium and Ti/diamine ratios. Those homogeneous complexes could be used as a heterogenized catalyst after anchoring on the supports, and the immobilized titanium catalyst retained the catalytic activities for several times in the recycled reactions without leaching. The carbon support containing titanium has exhibited superior activity to the silica support. Especially, when the titanium complex was anchored on the support which was fabricated by the hydrolysis of tripropylaminosilane itself, the resulting titanium catalyst showed the highest BD conversion and CDT selectivity.

Characterization and Cloning of a Phytase from Escherichia coli WC7. (Escherichia coli WC7가 생산하는 Phytase의 효소특성과 그 유전자의 클로닝)

  • 최원찬;오병철;김형권;강선철;오태광
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.1-7
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    • 2002
  • Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

Synthesis of trifluoromethylated dihydro-1,4-dithiin carboxamides and their antifungal activities (Trifluoromethylated Dihydro-1,4-dithiin carboxanilide 유도체의 합성 및 살균활성)

  • Hahn, Hoh-Gyu;Nam, Kee-Dal;Chang, Kee-Hyuk;Lee, Seon-Woo;Cho, Kwang-Yun
    • The Korean Journal of Pesticide Science
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    • v.5 no.2
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    • pp.26-32
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    • 2001
  • [ ${\alpha},{\beta}$ ]-Unsaturated carboxamides 12 with trifluromethylated dihydro-1,4-dithiins were synthesized for the purpose of development of new agrochemical fungicide. Chlorination of trifluoromethylated ${\beta}$-ketoester 4 followed by tile reaction with 1,2-ethanedithiol gave intermediate 1,4-dithiane 9. Without purification of 9 substitution of hydroxy by chlorine followed by dehydrochlorination in the presence of triethylamine afforded trifluoromethylated dihydro-1,4-dithiin ethyl ester 7. Activation of the hydroxy of the carboxylic acid 10 obtained from the hydrolysis of 7 and then reacted with various amines gave the corresponding trifluoromethylated dihydro-1,4-dithiin carboxamides. Antifungal screening (in vivo) against typical plant diseases, Rice Blast, Rice Sheath Blight, Cucumber Gray Mold, Tomato Late Blight, Wheat Leaf Rust, and Barley Powdery Mildew of the synthesized compounds was carried out. As a result, most of the compounds shlowed weak antifungal activities and some compounds in which isopropyl group was substituted in meta of the phenyl showed antifungal activity (99%) at 250 ppm against the disease Wheat Leaf Rust.

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Production of Xylooligo-Saccharides and Purification of Extracellular Xylanase from Streptomyces chibaensis J-59 (방선균 Streptomyces chibaensis J-59 Xylanase의 정제 및 자일로 올리고당(Xylooligo-Saccharides)의 생산)

  • Joo, Gil-Jae;Rhee, In-Koo
    • Current Research on Agriculture and Life Sciences
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    • v.14
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    • pp.111-122
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    • 1996
  • S. chibaensis J-59 produced an extracellular xylanase in a CSL medium composed of 1.5% com steep liquor, 0.1% $MgSO_4{\cdot}7H_2O$, 0.012% $CoCl_2{\cdot}6H_2O$, and 0.15% glucose containing xylan. but it did not produce in the culture medium containing xylose. The production of enzyme reached to a maximum level (0.83 uints/ml) when bacteria were cultured in 2.5 l jar fermentor for 48hrs at $30^{\circ}C$ and pH 7.0. Furthermore, S. chibaensis J-59 produced an intracellular glucose isomerase in a medium containing xylan and/or xylose. Xylanase was purified 29-fold over the culture supernatants of S. chibaensis J-59 by ammonium sulfate fractionation, chromatography on DEAE-Sephadex A-50, and gel filtration on Sephadex G-200. The purified enzyme is a monomeric enzyme with a native molecular mass of 25 kDa and a subunit molecular mass of 25 kDa. The purified enzyme requires $Mg^{2+}$ for activity, $Ca^{2+}$, $Co^{2+}$ is not an inhibitor but inhibit by $Fe^{3+}$, $Hg^{2+}$, and $Cu^{2+}$, sodium dodecyl sulfate, N-bromosuccinide. Pattern of hydrolysis demonstrated that the xylanase was an endo-splitting enzyme able to break down birchwood xylan at random giving xylobiose, xylotriose and xylotetrose as the main end products.

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Characteristics of protein from red crab (Chionoecetes japonicus) shell by commercial proteases (효소적 가수분해에 의한 홍게껍질 단백질의 특성)

  • Noh, Kyung-Hee;Min, Kwan-Hee;Seo, Bo-Young;Kim, So-Hee;Seo, Young-Wan;Song, Young-Sun
    • Journal of Nutrition and Health
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    • v.45 no.5
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    • pp.429-436
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    • 2012
  • This study was performed to examine the characteristics of protein of red crab (Chionoecetes japonicus) shell powder hydrolyzed by commercial proteases. Red crab shell was digested by commercial proteases, such as Protamex (P), Neutrase (N), Flavourzyme (F), Alcalase (A), Protease M (PM) and Protease A (PA). Protein yield analyzed by Biuret assay, absorbance at 280 nm and brix revealed that PA was the enzyme having the highest proteolytic activity. SDS PAGE showed that molecular weight of proteins produced by protease treatments was various and below 150 kDa. Combinational treatment of proteases (PA + P, PA + PM, PA + F, PA + A) was tried whether these increase protein hydrolysis from red crab shell powder compared to a PA single treatment. Soluble protein content was similar, but amino acid concentration by combinational treatments was higher than PA single treatment [PA + P 247.4 mg/g > PA + F (206.4 mg/g) > PA + A (133.4 mg/g) > PA + PM (59.1 mg/g) > PA (54.9 mg/g)]. Amino acid composition by combinational treatments was slightly different. Most abundant essential amino acids were phenylalanine, glycine, alanine, and leucine, whereas tyrosine and cystine were not detected.

Protein Quality Evaluation and Effect of Plasma Lipid Contents of Acid Hydrolysates of Cocoon inn Rats Fed by High Cholesterol, High Triglyceride and High Sucrose Diet (누에고치 산 가수분해물의 단백질의 질적 평가와 고콜레스테롤, 고지질, 고당질식이 흰쥐의 혈장지질에 미치는 영향)

  • 황은희;강병기;김복량;이형자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.1004-1009
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    • 2001
  • Acid hydrolysates of cocoon was gained by acid hydrolysis of 2 N HCl, 11$0^{\circ}C$, 48 hours, neutralization and desalting from the cocoon. The amino acid compositions of acid hydrolysates of cocoon were glycine 43.25%, alanine 34.39%, serine 10.05% and valine 2.44%. The contents of essential amino acid was 10.05%. Food efficiency ratio of acid hydrolysates of cocoon group was equal to the reference protein, casein. Liver weight, GOT, GPT activity, serum albumin and serum total protein level of rats were not significantly different among the experimental groups. Therefore, the protein acid hydrolysates of cocoon is not of high quality. When the rat fed with high cholesterol, high lipid, and high sucrose diet was administered with 5% acid hydrolysates of cocoon, its plasma lipids concentration of acid hydrolysates of cocoon was favorably affected: its triglyceride was decreased, and the level of phospholipid and HDL cholesterol were increased. There was also an unfavorable effect: the levels of LDL cholesterol and total cholesterol went up. Therefore, the acid hydrolysates of cocoon is not a good protein food source, but is can be used a cosmetic, medical, or packing material. Further research will reveal how it will affect or improve plasma lipid.

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THE AFFINITY OF CALMODULIN-AFFIGEL FOR INOSITOL TRIPHOSPHATE KINASE FROM BOVINE BRAIN (소의 뇌 Inositol triphosphate kinase와 Calmodulin-Affigel과의 친화도)

  • Lim, Sung-Woo;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.7 no.1
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    • pp.39-50
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    • 1990
  • The one event on signalling mechanism is the cleavage by adenyl cyclase of ATP into second messenger, cyclic AMP. The other transfer system of inositol metabolism. it is widely recognized that hydrolysis of the minor membrane lipid phosphoinositide bisphosphate($PIP_2$) initiated by occupation of certain receptors and catalyzed by phospholipase C, lead to toe generation of the two intracellular messengers, inositol triphosphate($IP_3$) and diacylglycerol(DG). $IP_3$ is converted to inositol tetrakisphosphate($IP_4$) by $IP_3$ kinase. In the present study, it is that purification of calmodulin is used by phenyl-Sepharose CL-4B chromatography. it's molecular weigh, 17.000 in SDS-polyacrylamide gel electrophoresis. In order to observe the affinity between calmodulin (CaM)-Affigel 15 and $IP_3$ kinase, and isolated $IP_3$ kinase, was applied in CaM-Affigel with $Ca^{2+}$ equilibirum buffer and EGTA equilibirum buffer. We compared with binding and elution effect of $IP_3$ kinase in several condition of buffer. In affinity of binding. $Ca^{2+}$ equilibrium buffer was in the most proper condition. and elution, CaM/$Ca^{2+}$ buffer(CE1 10.36, CE2 12. 76pM/min/mg of protein) was effected much more than EGTA buffer(E2 1.48, E3 2.43pM/min/mg of protein), but CaM/$Ca^{2+}$ stimulate the activity of $IP_3$ kinase. And then, several detergents such as sodium deoxycholate, tween 20. cholic acid, polyethylene glycol, chaps were applied. The 0.2% chaps buffer(E2 23.19, E3 8.05pM/min/mg of protein) was the most effective in elution of $IP_3$ kinase.

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Soil Carbon and Microbial Activity Influenced by Pasture and Rice Paddy Management (목초재배지 및 벼논 관리 변화에 따른 토양 탄소 및 미생물 활성도)

  • Yoo, Ga-Young;Kim, Hyun-Jin;Kim, Ye-Sol;Jung, Min-Hung
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.3
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    • pp.435-443
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    • 2012
  • This study investigated soil carbon storage and microbial activities influenced by different management practices in rice paddies and pastures. Soils under a single-crop farming of rice (CON) and rice-Italian ryegrass rotation farming (IRG) were compared in Jangheung, Jeollanam-do, Seocheon and Cheonan, Chungcheongnam-do. Soils from pastures were analyzed to investigate the effect of duration period (P1, P2, P3) in Namwon, Jeollabuk-do and Seosan, Chungcheongnam-do. In rice paddy, total and particulate carbon (PC) concentrations in the IRG soils were significantly higher than those in the CON soils both in Jangheung and Seocheon where the IRG has been established for three years, whereas carbon concentrations were not significantly different in Cheonan where IRG planting history is only one year. In rice paddy soils, PC was suggested as an early indicator to monitor changes in soil carbon storage followed by adopting different management practices. In pasture, total and PC concentrations increased with duration period especially in the 0-5 cm soils. Contrary to the rice paddy soils, the magnitude of change in PC concentration is not as great as that in total carbon concentration, implying that there is a need to develop a new early indicator other than PC using different fractionation scheme. The soil carbon storage in pasture also increased with years since establishment and the increasing rate was significantly greater in the early stage (0-5 yrs) than the later one (> 5 yrs). Microbial activities measured from fluorescein diacetate (FDA) hydrolysis analysis were significantly lower in the IRG soils than CON soils, whereas no difference was observed in the pastures of different ages. This shows that FDA activity is not a sensitive indicator to differentiate soil qualities influenced by management practices if it is used by itself.