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首页> 外文期刊>Biotechnology Progress >Effect of Enzyme Dehydration on Alcalase-Catalyzed Dipeptide Synthesis in Near-Anhydrous Organic Media
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Effect of Enzyme Dehydration on Alcalase-Catalyzed Dipeptide Synthesis in Near-Anhydrous Organic Media

机译：酶脱水对近水有机介质中碱性磷酸酶催化二肽合成的影响

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摘要

The effect of enzyme dehydration by molecular sieves on the coupling of phenylalanine amide and the carbamoylmethyl ester of N-protected phenylalanine in near-anhydrous tetra-hydrofuran was investigated. This coupling was catalyzed by Alcalase covalently immobilized onto macroporous acrylic beads (Cov); these immobilized enzymes were hydrated prior to use. The dehydration kinetics of Cov by molecular sieve powder were determined by incubating Cov with different amounts of molecular sieve powder for different periods of time (0-80 h). Subsequently, the remaining coupling activity of Cov was measured. Dehydration-induced inactivation of Cov by molecular sieve powder was found to occur in three phases: (1) an initial, rapid, major dehydration-induced inactivation that takes place during the first activity measurement, (2) a phase of first-order inactivation, and (3) a plateau phase in activity. These dehydration kinetics were incorporated into a previously found reaction kinetics model. The resulting model was then used to fit progress curve data of the coupling in the presence of different amounts of molecular sieve powder. Upon establishment of parameter values, the model was used to predict independent data sets and found to work well.
机译：研究了分子筛酶脱水对近无水四氢呋喃中苯丙氨酸酰胺与N-保护的苯丙氨酸的氨基甲酰基甲基酯偶联的影响。这种偶联是通过共价固定在大孔丙烯酸珠（Cov）上的Alcalase催化的。这些固定化的酶在使用前需要水合。通过用不同量的分子筛粉孵育Cov不同时间（0-80小时），确定分子筛粉对Cov的脱水动力学。随后，测量Cov的剩余偶联活性。发现分子筛粉末可导致脱水诱导的Cov失活分为三个阶段：（1）在首次活性测量期间发生了初始的，快速的，主要的脱水诱导的失活;（2）一级失活的阶段。 ，以及（3）活动处于平稳阶段。这些脱水动力学被并入先前发现的反应动力学模型中。然后，在存在不同量的分子筛粉末的情况下，将所得模型用于拟合偶合的进度曲线数据。建立参数值后，该模型可用于预测独立的数据集，并能很好地工作。

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