• Title/Summary/Keyword: CBH

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Optimized Conditions for In Situ Immobilization of Lipase in Aldehyde-silica Packed Columns

  • Seo Woo Yong;Lee Kisay
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.6
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    • pp.465-470
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    • 2004
  • Optimal conditions for the in situ immobilization of lipase in aldehyde-silica packed columns, via reductive amination, were investigated. A reactant mixture, containing lipase and sodium borohydride (NaCBH), was recirculated through an aldehyde-silica packed column, such that the covalent bonding of the lipase, via amination between the amine group of the enzyme and the aldehyde terminal of the silica, and the reduction of the resulting imine group by NaCBH, could occur inside the bed, in situ. Mobile phase conditions in the ranges of pH $7.0{\~}7.8$, temperatures between $22{\~}28^{circ}C$ and flow rates from $0.8{\~}1.5\;BV/min$ were found to be optimal for the in situ immobilization, which routinely resulted in an immobilization of more than 70 mg­lipase/g-silica. Also, the optimal ratio and concentration for feed reactants in the in situ immobilization: mass ratio [NaCBH]/[lipase] of 0.3, at NaCBH and lipase concentrations of 0.75 and 2.5 g/L, respectively, were found to display the best immobilization characteristics for concentrations of up to 80 mg-lipase/g-silica, which was more than a 2-fold increase in immobilization compared to that obtained by batch immobilization. For tributyrin hydrolysis, the in situ immobilized lipase displayed lower activity per unit mass of enzyme than the batch-immobilized or free lipase, while allowing more than a $45\%$ increase in lipase activity per unit mass of silica compared to batch immobilization, because the quantity of the immobilization on silica was aug­mented by the in situ immobilization methodology used in this study.

Purification and Structure Determination of the GTase Inhibitor from Cacao Bean Husk Extract (Cacao Bean Husk 추출물로부터 Glucosyltransferase 저해 활성 물질의 분리 및 구조 동정)

  • 권익부;안봉전이신영
    • KSBB Journal
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    • v.11 no.5
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    • pp.536-542
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    • 1996
  • The isolation of active compounds showing the inhibitory effect on glucosyltransferase(GTase) from cacao bean husk(CBH) extract was carried out for screening of anti-plaque agents. These active compounds were purified by additional column chromatography of MCI-gel CHP-20 and Sephadex LH-20 and their chemical structures were determined by NMR and mass spectroscopy. Two compounds showing the inhibitory effect on GTase from CBH extract were obtained. These compounds showed positive reactions with anisaldehyde-H2SO4 solution and FeCl3, and were identified as dimeric flavan-3-ols on TLC. By NMR and MS data analyses, the structures of two different flavan-3-ols were identified as procyanidin B-1 and procyanidin B-3, respectively.

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Comparison of Growth, Yield and Quality by Green Crop Treatments in Rice (Oryza sativa L.) Organic Cultivation (벼 유기재배 시 녹비작물 처리에 의한 작물의 생육, 수량 및 품질 비교)

  • Cha, Kwang-Hong;Oh, Hwan-Jung;Park, Heung-Gyu;An, Kyu-Nam;Park, Ro-Dong;Jung, Woo-Jin
    • Korean Journal of Organic Agriculture
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    • v.19 no.1
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    • pp.55-64
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    • 2011
  • To investigate a comparison of rice yield and quality by a utilization of green crops using barley and hairyvetch, this study was carried out in rice (Oryza sativa L.) organic cultivation field. Convention barley harvest (CBH), Green barley crop (GBC), Green barley+Green Hairyvetch crop (GB+HVC), and Non-Green crop (NGC) were treated in rice cultivation field. The results obtained as following: 1) Culm length and pancile length in Convention barley harvest (CBH) cultivation were longer than organic cultivation but were not significant difference in all treatments. Yields of milled rice in Green barley crop (GBC) and Green barley+Green Hairyvetch crop (GB+HVC) were by 90.3% and 95.9%, respectively. 2) Protein contents in Green barley crop (GBC) and Green barley+Green Hairyvetch crop (GB+HVC) were by 5.7% lower than 6.6% in Convention barley harvest (CBH) cultivation. Amylose contents in all treatment were similar as range 18.8% to 19.1%. Whiteness contents in Green barley crop (GBC) and Green barley+Green Hairyvetch crop (GB+HVC) were slightly higher by 38.9% and 39.1%, respectively, than 37.7% in Convention barley harvest (CBH) cultivation. 3) Palatability values in Barley harvest (BH) and Non-Green crop (NGC) were slightly higher by 82.0 and 83.8, respectively, than 77.6 in Convention barley harvest (CBH) cultivation. 4) Head rice in Non-Green crop (NGC) was 95.5%, while that of Convention barley harvest (CBH), Green barley crop (GBC), and Green barley+Green Hairy-vetch crop (GB+HVC) were slightly low as range 93.8% to 94.2%. White core and belly rice in Convention barley harvest (CBH) cultivation was the highest level by 1.7%. 5) Leaf blast, neck blast, sheath blight, rice stem maggot, rice leaf roller, rice leaf-tier, and green rice leafhopper were occurred lightly in rice field.

Production of Cellulases by Rhizopus stolonifer from Glucose-Containing Media Based on the Regulation of Transcriptional Regulator CRE

  • Zhang, Yingying;Tang, Bin;Du, Guocheng
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.514-523
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    • 2017
  • Carbon catabolite repression is a crucial regulation mechanism in microorganisms, but its characteristic in Rhizopus is still unclear. We extracted a carbon regulation gene, cre, that encoded a carbon catabolite repressor protein (CRE) from Rhizopus stolonifer TP-02, and studied the regulation of CRE by real-time qPCR. CRE responded to glucose in a certain range, where it could significantly regulate part of the cellulase genes (eg, bg, and cbh2) without cbh1. In the comparison of the response of cre and four cellulase genes to carboxymethylcellulose sodium and a simple carbon source (lactose), the effect of CRE was only related to the concentration of reducing sugars. By regulating the reducing sugars to range from 0.4% to 0.6%, a glucose-containing medium with lactose as the inducer could effectively induce cellulases without the repression of CRE. This regulation method could potentially reduce the cost of enzymes produced in industries and provide a possible solution to achieve the largescale synthesis of cellulases.

Comparative effects of dietary functional nutrients on growth performance, meat quality, immune responses, and stress biomarkers in broiler chickens raised under heat stress conditions

  • Kim, Deok Yun;Kim, Jong Hyuk;Choi, Won Jun;Han, Gi Ppeum;Kil, Dong Yong
    • Animal Bioscience
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    • v.34 no.11
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    • pp.1839-1848
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    • 2021
  • Objective: The objective of the present study was to investigate the comparative effects of dietary functional nutrients including glutamine (Gln), chromium picolinate (Cr picolinate), vitamin C (Vit C), betaine (Bet), and taurine (Tau) on growth performance, meat quality, immune responses, and stress biomarkers in broiler chickens raised under heat stress conditions. Methods: A total of 420 21-d-old Ross 308 male broiler chickens (initial body weight = 866±61.9 g) were randomly allotted to 1 of 7 treatment groups with 6 replicates. One group was kept under thermoneutral conditions and was fed a basal diet (PC, positive control). Other 6 groups were exposed to a cyclic heat stress condition. One of the 6 groups was fed the basal diet (NC, negative control), whereas 5 other groups were fed the basal diet supplemented with 0.5% Gln, 500 ppb Cr picolinate, 250 mg/kg Vit C, 0.2% Bet, or 1.0% Tau. The diets and water were provided ad libitum for 21 d. Results: Broiler chickens in NC group had decreased (p<0.05) growth performance and immune responses measured based on cutaneous basophil hypersensitivity (CBH), but increased (p<0.05) stress responses measured based on feather corticosterone concentrations and blood heterophil:lymphocyte than those in PC group. However, none of dietary functional nutrients had a positive effect on growth performance of broiler chickens. Dietary supplementation of 250 mg/kg Vit C improved (p<0.05) CBH responses of broiler chickens, but other functional nutrients had no such an improvement in CBH responses. All functional nutrients decreased (p<0.05) stress responses of broiler chickens. Conclusion: Functional nutrients including Gln, Cr picolinate, Vit C, Bet, and Tau at the supplemental levels used in this study decrease stress responses of broiler chickens to a relatively similar extent. However, this reduction in stress responses could not fully ameliorate decreased productive performance of broiler chickens raised under the current heat stress conditions.

Enhancing Cellulase Production in Thermophilic Fungus Myceliophthora thermophila ATCC42464 by RNA Interference of cre1 Gene Expression

  • Yang, Fan;Gong, Yanfen;Liu, Gang;Zhao, Shengming;Wang, Juan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1101-1107
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    • 2015
  • The role of CRE1 in a thermophilic fungus, Myceliophthora thermophila ATCC42464, was studied using RNA interference. In the cre1-silenced strain C88, the filter paper hydrolyzing activity and β-1,4-endoglucanase activity were 3.76-, and 1.31-fold higher, respectively, than those in the parental strain when the strains were cultured in inducing medium for 6 days. The activities of β-1,4-exoglucanase and cellobiase were 2.64-, and 5.59-fold higher, respectively, than those in the parental strain when the strains were cultured for 5 days. Quantitative reverse-transcription polymerase chain reaction showed that the gene expression of egl3, cbh1, and cbh2 was significantly increased in transformant C88 compared with the wild-type strain. Therefore, our findings suggest the feasibility of improving cellulase production by modifying the regulator expression, and an attractive approach to increasing the total cellulase productivity in thermophilic fungi.

Cellulase Production in the Digestive Organs of Reticulitermes speratus, a Native Termite from Milyang, Korea

  • Lee, Young-Min;Kim, Yoon-Hee;Cho, Moon-Jung;Shin, Keum;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.5
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    • pp.421-428
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    • 2010
  • This study investigated on enzyme production in the digestive organs of the native termite (Reticulitermes speratus) in Milyang, Korea. Four types of major cellulases [EG (endo-1,4-${\beta}$-glucanase), BGL (${\beta}$-glucosidase), CBH (cellobiohydrolase) and BXL (${\beta}$-1,4-xylosidase)] were present in the digestive organs of the termite. The strong enzyme activity for BGL was found from the native termite, and also shown that the enzyme was distributed in the salivary gland, foregut, and hindgut. BXL, which breaks down hemicellulose near the amorphous region, was detected mainly from salivary gland, foregut, and midgut. However, CBH was distributed mainly in the hindgut. Meanwhile, EG which degrades cellulose, was found mainly in the hindgut and salivary glands. These facts indicate that celluases production patterns are differ from different sites compare to the same species found in Japan, suggesting that enzyme production in the digestive organs of termites is changed according to their habitats.

Prediction on the Chiral Behaviors of Drugs with Amine Moiety on the Chiral Cellobiohydrolase Stationary Phase Using a Partial Least Square Method

  • Choi, Sun-Ok;Lee, Seok-Ho;Park Choo , Hea-Young
    • Archives of Pharmacal Research
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    • v.27 no.10
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    • pp.1009-1015
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    • 2004
  • Quantitative Structure-Resolution Relationship (QSRR) using the Comparative Molecular Field Analysis (CoMFA) software was applied to predict the chromatographic behaviors of chiral drugs with an amine moiety on the chiral cellobiohydrolase (CBH) columns. As a result of the Quantitative CoMFA-Resolution Relationship study, using the partial least square method, prediction of the behavior of drugs with amine moiety upon chiral separation became possible from their three dimensional molecular structures. When a mixed mobile phase of 10 mM aqueous phosphate buffer (pH 7.0) - isopropanol (95 : 5) was employed, the best Quantitative CoMFA-Resolution Relationship, derived from the study, provided a cross-validated $q^2$ = 0.933, a normal $r^2$ = 0.995, while the best Quantitative CoMFA-Separation Factor Relationship, also derived from the study, yielded a cross-validated $q^2$ = 0.939, a normal $r^2$ = 0.991. When all of these results are considered, this QSRR-CoMFA analysis appears to be a very useful tool for the preliminary prediction on the chromatographic behaviors of drugs with an amine moiety inside chiral CBH columns.

Characterization of Cellulases from Schizophyllum commune for Hydrolysis of Cellulosic Biomass (Schizophyllum commune에 의한 Cellulase 생산 및 섬유소계 바이오매스의 당화를 위한 효소적 특성)

  • Kim, Hyun-Jung;Kim, Yoon-Hee;Cho, Moon-Jung;Shin, Keum;Lee, Dong-Heub;Kim, Tae-Jong;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.6
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    • pp.547-560
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    • 2010
  • The optimum culture condition of Schizophyllum commune for the cellulase production and its enzymatic characteristics for saccharification of cellulosic biomass were analyzed. S. commune secrets ${\beta}$-1,4-xylosidase (BXL) and cellulases, including endo-${\beta}$-1,4-glucanase (EG), cellobiohydrolase (CBH), and ${\beta}$-glucosidase (BGL). The optimum reaction temperature for all cellulases was $50^{\circ}C$ and the thermostable range was $30{\sim}40^{\circ}C$C. The optimum reaction pH for all cellulases was 5.5 in a range of temperature from $0^{\circ}C$ to $55^{\circ}C$. The best nutritions for the cellulase production of S. commune among tested nutrients were 2% cellulose for the carbon source and corn steep liquor or peptone/yeast extract for the nitrogen source without vitamins. The environmental culture condition for the cellulase production was 5.5~6.0 for pH at $25{\sim}30^{\circ}C$. The enzyme activities of EG, BGL, CBH, and BXL were 3670.5, 631.9, 398.5, and 15.2 U/$m{\ell}$, respectively, after concentration forty times from the culture broth of S. commune which was grown at the optimized culture condition. Alternative filter paper unit assay showed 11 FPU/$m{\ell}$ enzyme activity. The saccharification tests using cellulase of S. commune showed the low saccharification rate on tested hardwoods but a high value of 50.5% on cellulose, respectively. The saccharification rate (50.5%) of cellulose by cellulase produced in this work is higher than 45.7% in the commercial enzyme (Celluclast 1.5L, 30 FPU/g, glucan).

Enzyme Activities and Cellulose Degradation of Domestic Softwoods in Shaking Culture of Fomitopsis palustris (국내산 침엽수 목분의 진탕배양에서 나타난 Fomitopsis palustris의 효소 활성 및 셀룰로오스 분해)

  • Choi, Doo-Yeol;Lee, Young-Min;Kim, Young-kyoon;Yoon, Jeong-Jun;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.6
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    • pp.91-99
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    • 2007
  • Activities of the extracellular enzyme from Fomitopsis palustris, a brown-rot fungi, and by which crystallinity changes of cellulose in the various softwoods, such as Larix leptolepsis, Finns rigida, Finns koraiensis and Finns densiflora by liquid culture, were investigated. Activity of Cellobiohydrolase (CBH) from F. palustris was detected in the every test softwoods culture, showing activities of the Endoglucanase (EG), $\beta$-glucosidase (BGL) and $\beta$-1,4-xylosidase (BXL). It was shown high enzyme activities in the sapwood culture than heartwood of the same wood species, However, the enzyme activities in most of test wood cultures increased with longer incubation time, indicating a possibility of intermix sapwood and heartwood for degradation process by enzyme. Also it was shown that protein patterns of the extracellular enzyme from F. palustris in wood particle substrate of the several domestic softwoods were similar with each other wood species, which suggested the possibility of mixing all softwoods in saccharification by enzyme from F. palustris. Crystallinity reduction value of cellulose by F. palustris was 4.2~20.4% in 4 weeks cultivation, 12.9~28.9% in 8 weeks.