• Biotechnology and Bioprocess Engineering:BBE
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• 제9권6호
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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.

• KSBB Journal
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• 제11권5호
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• pp.536-542
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• 1996

충치 예방 물질의 탐색 연구 일환으로, Cacao bean husk(CBH) 추출물로부터 얻어진, gluco-syltransferase(Gtase) 저해 활성 분획을 분리하였 다. 이를 다시 MCI-gel CHP-20와 Sephadex LH 20의 column chromatography법으로 분리, 정제하였으며, 이로부터 얻어진 2개의 GTase 활성 저해 물질에 대한 구조 해석을 행하였다. 분리.정제된 2개의 GTase 저해 활성 물질은 anisaldehyde-$H_2SO_4$ 및 FeCb에서 각각 갈색 및 청색 반응을 하며, 또 TLC상에서 dimer의 flavan-3-ol $R_r$값을 나타내는 단일 물질이었다. NMR(PMR, C CMR) 빛 mass spectrum으로 두 정제 분획에 대 한 화학 구조를 분석한 결과, G Tase 저해능을 나타 내는 단물질은 (-)-epicatechin-($\beta\rightarrow8$)-catechin 으로 결합된 procyanidin B-1 ($C_{30}H_{26}O_{12}$) 및 cate chin의 2량체인 procyanidin B一3($C_{30}H_{26}O_{12}$)이었다.

• 한국유기농업학회지
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• 제19권1호
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• pp.55-64
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• 2011

벼 재배포장에서 녹비작물의 이용성을 조사하고자 보리와 헤어리베치를 적용하여 시험구를 배치하였다. 무농약 4년차 벼 재배포장에서 벼 수확 후 재배한 보리를 녹비작물로 이용한 처리구(보리녹비구), 보리와 헤어리베치를 혼파한 다음 녹비작물로 이용한 처리구(보리+헤어리베치 녹비구), 벼 수확 후 녹비작물을 재배하지 않은 방임구(나지방임구)와 보리를 재배한 후 수확한 처리구(관행구)에서 호품벼를 2009년 6월 15일 평당 60주를 기준으로 기계 이앙하여 얻어진 시험결과를 요약하면 다음과 같다. 1. 간장과 수장은 관행구가 조금 길었으며, 처리간에는 차이가 없었다. 백미 수량은 관행구를 100%로 볼 때 보리녹비구가 90.3%, 보리+헤어리베치 녹비구가 95.9%로 나타났다. 2. 벼 품질 평가에서 보리녹비구와 보리+헤어리베치 녹비구의 단백질 함량은 관행구 6.6%에 비해 5.7%로 낮았다. 아밀로스함량은 모든 처리구에서 18.8~19.1% 범위로 유사한 경향을 보였다. 백도는 관행구 37.7에 비해 보리녹비구 38.9와 보리+헤어리베치 녹비구 39.1로 약간 높게 나타났다. 3. 취반미 윤기치는 관행구 77.6에 비해 보리수확구가 82.0, 나지방임구가 83.8로 양호하게 나타났다. 4. 완전립율이 나지방임구는 95.5%인 반면에 관행구, 보리녹비구, 보리+헤어리베치 녹비구에서 93.8~94.2% 범위로 약간 낮게 나타났다. 쇄립율도 유사한 경향이었으며 분상질립율은 관행구가 1.7%로 가장 높았다. 5. 본 시험에서 발생된 병해는 잎도열병, 이삭도열병, 잎집무늬마름병이었다. 해충으로는 굴파리류, 혹명나방, 줄점팔랑나방, 끝동매미충이었다. 기상환경이 병해충발생에 불리한 조건으로 경과되어 병해충 발생이 전반적으로 적었으며 처리간에도 유의적인 차이를 나타내지 않았다.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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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.

• Animal Bioscience
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• 제34권11호
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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.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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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.

• Journal of the Korean Wood Science and Technology
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• 제38권5호
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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.

• Archives of Pharmacal Research
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• 제27권10호
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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.

• Journal of the Korean Wood Science and Technology
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• 제38권6호
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• pp.547-560
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• 2010

본 연구에서는 Schizophyllum commune의 당 분해효소 생산을 위한 최적 배양 조건과 목질바이오매스에 대한 당화 특성에 대하여 연구하였다. S. commune 균체 외 효소에는 endo-${\beta}$-1,4-glucanase (EG), cellobiohydrolase (CBH), ${\beta}$-glucosidase (BGL)와 같은 cellulase와 ${\beta}$-1,4-xylosidase (BXL)이 함유되어 있고 그 중에서 EG 및 BGL활성이 비교적 높은 활성을 나타낸 것으로 밝혀졌다. S. commune에서 생산된 EG, BGL, 및 CBH의 최적 온도는 $50^{\circ}C$이었으나, 열안정성을 가지는 온도범위는 $30{\sim}40^{\circ}C$였다. 그리고 최적 pH는 5.5이었으며 열 안정성을 나타내는 온도범위에서의 적정 pH는 동일한 pH 5.5이었다. Cellulase 생산을 위한 S. commune의 최적배양 조건은, 탄소원으로 천연 cellulose, 질소원으로는 corn steep, 또는 peptone/yeast extract 혼합물, 비타민은 첨가하지 않는 것이 cellulase 효소활성 증가에 적절한 것으로 밝혀졌다. 또한 탄소원의 최적 첨가 농도는 2% (w/w), 적정 배양 pH 및 온도는 5.5~6.0과 $25{\sim}30^{\circ}C$로 밝혀졌다. 본 연구에서 도출된 최적 배양 조건으로 S. commune를 배양시키고 40배로 농축한 결과, EG가 3670.5 U/$m{\ell}$, BGL과 CBH가 각각 631.9 U/$m{\ell}$, 398.5 U/$m{\ell}$, BXL이 15.2 U/$m{\ell}$로 매우 높은 효소 활성을 나타냈다. 동일한 효소의 Filter Paper Unit도 11 FPU/$m{\ell}$로 상당히 높았다. 최적배양조건에서 얻어진 S. commune 효소로 다양한 기질에 대해 당화 시험을 실행한 결과, 전처리를 하지 않은 공시 활엽수에 대하여 낮은 당화율을 나타냈으나 천연 cellulose (Aldrich, ~20 micron) 및 볏짚의 경우에는 각각 50.5% 및 33.1%의 높은 당화성능을 나타냈다. 이 같은 당화 수준은 동일 효소농도 (30 FPU/g, glucan)로 비교했을 때 Trichoderma reesei 유래 상용화 효소인 Celluclast 1.5L의 약 110% 수준을 나타냄으로써, 대량생산기술 개발과정을 통해 목질계 당화 효소로의 상용화 가능성이 높은 균주로 평가되었다.

• Journal of the Korean Wood Science and Technology
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• 제35권6호
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• pp.91-99
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• 2007

소나무, 낙엽송, 리기다소나무, 잣나무 등 국내산 침엽수 목분을 기질로 진탕배양했을 때 갈색부후균인 Fomitopsis palustris로부터 분비된 당 분해성 균체 외 효소는 Endoglucanase (EG), $\beta$-glucosidase (BGL) 및 $\beta$-1,4-xylosidase (BXL)와 함께 결정형 셀룰로오스를 분해하는 Cellobiohydrolase (CBH)도 활성을 갖은 것으로 나타났다. 4주간 배양에서 변재는 심재에 비해 큰 효소활성을 나타냈으나, 배양기간을 증가시킴으로써 효소활성이 커지는 것으로 밝혀져 효소에 의한 목질바이오매스 분해의 경우 심변재 혼합처리도 가능할 수 있음이 시사되었다. 그리고, 균체 의 단백질을 SDS-PAGE로 분석한 결과, 대부분의 수종에서 나타나는 효소의 단백질 패턴은 거의 유사한 것으로 나타나 효소를 이용한 목질계 바이오매스 분해의 경구 수종별 혼합처리도 가능함이 시사되었다. F. palustris에 의해 분해된 목분(60 mesh 통과) 셀룰로오스의 결정화도 감소율은 4주 배양에서 약 4.2~20.4%, 8주 배양에서 약 12.9~28.9% 수준으로 나타났다.