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首页> 外文期刊>Analytical Sciences >Analytical Methods for the Detection and Purification of ε-Poly-L-lysine for Studying Biopolymer Synthetases, and Bioelectroanalysis Methods for Its Functional Evaluation
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Analytical Methods for the Detection and Purification of ε-Poly-L-lysine for Studying Biopolymer Synthetases, and Bioelectroanalysis Methods for Its Functional Evaluation

机译:用于研究生物聚合物合成酶的ε-聚L-赖氨酸检测和纯化的分析方法,以及其功能评价的生物电解分析方法

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摘要

This article describes new analytical methods for studying biopolymer ε-poly-L-lysine (εPL). The produced amount of εPL in culture broth can be determined based on the precipitation of polycationic εPL with a colored heteropolymolybdate anion and the color change of the supernatant. The product can be separated and purified by precipitation with the tetraphenylborate anion and reprecipitation in the form of the hydrochloride salt. These methods have been applied advantageously to the screening of εPL-synthetase. Also, pyrophosphate can be determined colorimetrically based on the formation of 18-molybdopyrophosphate species. The pyrophosphate determination has been successfully applied to the assay of adenylation enzyme, which plays important roles in the biosynthetic mechanism. Under certain conditions, εPL associates with a redox enzyme, glucose oxidase. The effect of the adduction on the stability and reaction rate of the enzyme can be evaluated by measuring the bioelectrocatalytic current, which is related to the enzyme activity. Electrochemical studies showed new applications of εPL as an enzyme stabilizer and a reaction enhancer.
机译:本文介绍了用于研究生物聚合物ε-聚L-赖氨酸(εpl)的新分析方法。培养肉汤中产生的εpl的产生量可以基于具有有色杂多元斑阴离子和上清液的颜色变化的聚阳离子εpl的沉淀来确定。通过用四苯基硼酸盐的阴离子和盐酸盐形式的沉淀分离和纯化产物。这些方法已经有利地应用于εpl-合成酶的筛选。而且,焦磷酸盐可以基于18摩尔磷酸盐物质的形成比例。焦磷酸盐测定已成功应用于腺苷酸化酶的测定,这在生物合成机制中起重要作用。在某些条件下,εpl与氧化还原酶,葡萄糖氧化酶联系起来。相应对酶的稳定性和反应速率的影响可以通过测量与酶活性有关的生物来催化电流来评估。电化学研究表明,εpl作为酶稳定剂和反应增强剂的新应用。

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