• 한국미생물·생명공학회지
• /
• 제23권4호
• /
• pp.425-431
• /
• 1995

A kinetic model of the cyclodextrin formation in a heterogeneous enzyme reaction system using swollen extrusion starch as substrate was derived emphasing the structural features of extrusion starch. The degree of gelatinization, the ratio of accessible and inaccessible portion of extrusion starch, adsorption of CGTase on swollen starch, the structural transformation during reaction, and product inhibition caused by produced CDs were considered in deriving kinetic model. Various kinetic constants were also evaluated. The derived kinetic equation was numerically simulated, which result showed that the derived kinetic equations can be used to predict the experimental data reasonably well under the various experimental conditions. Kinetic model can be utilized for the optimization of enzyme reactor and the process development for CD production from swollen extrusion starch.

• Bulletin of the Korean Chemical Society
• /
• 제26권4호
• /
• pp.515-522
• /
• 2005

Enzyme-metal combo catalysis is described as a useful methodology for the synthesis of optically active compounds. The key point of the method is the use of enzyme and metal in combination as the catalysts for the complete transformation of racemic substrates to single enantiomeric products through dynamic kinetic resolution (DKR). In this approach, enzyme acts as an enantioselective resolving catalyst and metal does as a racemizing catalyst for the efficient DKR. Three kinds of enzyme-metal combinations - lipase-ruthenium, subtilisin-ruthenium, and lipase-palladium –have been developed as the catalysts for the DKRs of racemic alcohols, esters, and amines. The scope of the combination catalysts can be extended to the asymmetric transformations of ketones, enol acetates, and ketoximes via the DKRs. In most cases studied, enzyme-metal combo catalysis provided enantiomerically-enriched products in high yields.

• Archives of Pharmacal Research
• /
• 제22권1호
• /
• pp.13-17
• /
• 1999

The kinetic constants and the reaction mechanism of the K228G mutant horse liver alcohol dehyrogenase isoenzyme E (HLADH-E) were compared to the wild-type enzyme. All the Km and Ki constants of the mutant enzyme for NAD+, ethanol, acetaldehyde and NADH were larger than those of the wild-type enzyme. The dissociation constants for the NADH and $NAD^{+}$ (Kiq and Kia) were greatly increased by 130-and 460-fold, respectively. The product inhibition patterns suggested that the reaction mechanism of the mutant enzyme was changed to Random Bi Bi. These results could attribute to the increase in the dissociation rate of coenzyme with the substitution at Lys-228 residue.

• Bulletin of the Korean Chemical Society
• /
• 제28권11호
• /
• pp.2096-2098
• /
• 2007

• KSBB Journal
• /
• 제10권5호
• /
• pp.540-546
• /
• 1995

bacterial alkaline protease를 이 용한 corn glu ten meal의 효소가수분해시 pH, 온도, 효소대 기질의 질량비율 등에 따른 반응특성 및 반응의 적정화 를 꾀하였고, 효소의 deactivation여부에 초점을 맞추어 효소반응속도론적 방정식을 제안조사하였다. 그 결과,$50^{\circ}C$, pH 9~10에서 가장 높은 가수분해 도를 나타내었고 $e_0/s_o$가 높을수록 반응속도 빛 최종 가수분해도가 증가하였다. 최종가수분해도는 gluten meal전체질량기준으로 17~20%, gluten meal내의 단백질질량기준으로 25 ~ 28 % 였다. 반응후반에서의 반응속도감소의 주된 원인은 기 질소진 (substrate de pletion) 이며 이때 enzyme deactivation 및 product inhibition의 영향은 미미한 것으로 확인되었다. 효소 deactivation항을 무시하여 변형시킨 model equa-tlOn에 의해 이 효소반응에 해당하는 여러 kinetic parameter들의 값을 계산분석한 결과 product inhi bition효과가 미미함을 확인하였다. 변형된 kinetic equation과 실험적으로 얻은 가수분해 데이타는 거 의 완벽하게 일치하였다. 효소반응을 $100\times$scale­u up한 결과 가수분해 profile 및 가수분해도는 $1\times$ 와 비교시 거의 일치하였으므로 이 효소반응이 용이하 게 scale-up될 수 있음을 확인하였고, 아미노산분석 결과 가수분해액을 미생물발효기질용 질소원으로 사 용할 수 있음을 제시하였다.

• 한국미생물생명공학회:학술대회논문집
• /
• 한국미생물생명공학회 1986년도 추계학술대회
• /
• pp.510.1-510
• /
• 1986

Many enzymes require the participation of readily dissociable coenzymes as NAD for thir catalytic activities. The continuous utilization of the enzymes requires the retention and regeneration of the coenzymes. For this purpose, several kinds of macromolecular NAD derivatives have been prepared by covalently attaching NAD to watersoluble polymers. We have prepared poly (ethylene glycol)-bound NAD (PEG-NAD) by coupling N$\^$6/-(2-carboxyethyl)-NAD to one terminal of ${\gamma}$ $\omega$-diaminoly (ethylene glycol) (Mr 3000) with water-soluble carbodiimide. PED-NAD thus obtained has one NAD moiety located at a terminal of the linear, flexible and hydrophilic chain of poly (ethylene glycol). PED-NAD has good coenzyme activity for various dehydrogenases and is applicable in a continuous enzyme reactor. To use these macromolecular NAD derivatives in an enzyme reactor, it si necessary to understand the behavior of the system in which the reactions of dehydrogenases are coupled by the recycling of the NAD derivative. We investigated the kinetic properties of a continuous enzyme reactor containing lactate dehydrogenase, alcohol dehydrogenase and PEG-NAD. The steady-state behavior of the enzyme reactor is explained by a simple kinetic model.

• 한국미생물·생명공학회지
• /
• 제17권6호
• /
• pp.552-555
• /
• 1989

Corynebacterium glutamicum의 NADPH-specific glutamate dehydrogenase를 이용하여 NADPH, NH$_4$Cl, $\alpha$-ketoglutarate의 기질에 대한 kinetics를 고찰하였다. 이들의 kinetic constants를 측정함으로서 정반응에로의 효소반응 기작은 첫번째 효소와 반응하는 기질이 NADPH 임을 확인할 수 있었다. Glutamate dehydrogenase 활성의 조절을 위한 metabolites의 효과를 고찰하여 본 결과 malate와 citrate 만이 효소에 억제 효과를 나타내었으며, potassium chloride는 효소활성에 가장 많은 영향을 주었다.

• 공업화학
• /
• 제17권6호
• /
• pp.658-664
• /
• 2006

$\rho$, $\alpha$-dimethyl benzyl alcohol을 효과적으로 분할하기 위하여 2상 동적 속도론적 광학분할(biphasic dynamic kinetic resolution, DKR)반응을 실시하였다. 라세미화 반응을 위하여 촉매로 산성 제올라이트를 사용하였고 속도론적 광학분할(kinetic resolution, KR)을 위하여 고정화 효소를 촉매로 사용하였다. 유기용매와 물을 용매로 사용하는 이상 DKR 반응에서, acyl donor, 반응온도, 기질의 농도, 두 가지 촉매의 상대적 비율 및 교반속도 등의 공정변수를 변화시켜가면서 DKR반응의 전환율과 생성물의 광학순도에 미치는 영향을 조사하였다. 그 결과, $\rho$, $\alpha$-dimethyl benzyl alcohol의 DKR 반응에서 99% 이상의 광학순도를 가지는 생성물을 최대 88%의 높은 수율로 얻을 수 있었으며, 높은 TON에서도 반응의 효율성이 유지되었고 촉매의 재사용 시에도 지속적인 활성을 나타내었다.

• BMB Reports
• /
• 제37권5호
• /
• pp.533-537
• /
• 2004

Previously published kinetic data on the interactions of seventeen different enzymes with their physiological substrates are re-examined in order to understand the connection between ground state binding energy and transition state stabilization of the enzyme-catalyzed reactions. When the substrate ground state binding energies are normalized by the substrate molar volumes, binding of the substrate to the enzyme active site may be thought of as an energy concentration interaction; that is, binding of the substrate ground state brings in a certain concentration of energy. When kinetic data of the enzyme/substrate interactions are analyzed from this point of view, the following relationships are discovered: 1) smaller substrates possess more binding energy concentrations than do larger substrates with the effect dropping off exponentially, 2) larger enzymes (relative to substrate size) bind both the ground and transition states more tightly than smaller enzymes, and 3) high substrate ground state binding energy concentration is associated with greater reaction transition state stabilization. It is proposed that these observations are inconsistent with the conventional (Haldane) view of enzyme catalysis and are better reconciled with the shifting specificity model for enzyme catalysis.

• Bulletin of the Korean Chemical Society
• /
• 제32권5호
• /
• pp.1500-1506
• /
• 2011

Tyrosinase is a widespread enzyme with great promising capabilities. The Lineweaver-Burk plots of the catecholase reactions showed that the kinetics of mushroom tyrosinase (MT), activated by $Co^{2+}$ and $Zn^{2+}$ at different pHs (6, 7, 8 and 9) obeyed the non-essential activation mode. The binding of metal ions to the enzyme increases the maximum velocity of the enzyme due to an increase in the enzyme catalytic constant ($k_{cat}$). From the kinetic analysis, dissociation constants of the activator from the enzyme-metal ion complex ($K_a$) were obtained as $5{\times}10^4M^{-1}$ and $8.33{\times}10^3M^{-1}$ for $Co^{2+}$ and $Zn^{2+}$ at pH 9 and 6 respectively. The structural analysis of MT through circular dichroism (CD) and intensive fluorescence spectra revealed that the conformational stability of the enzyme in these pHs reaches its maximum value in the presence of each of the two metal ions.