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优化影响磁性共聚物苯乙烯-二乙烯基苯合成的参数,以将其用作固定脂肪酶的有效基质。

Optimization of the parameters that affect the synthesis of magnetic copolymer styrene-divinilbezene to be used as efficient matrix for immobilizing lipases.

机构信息

Department of Chemical Engineering, Engineering School of Lorena, University of São Paulo, Lorena, SP, 12602-810, Brazil.

出版信息

World J Microbiol Biotechnol. 2018 Nov 7;34(11):169. doi: 10.1007/s11274-018-2553-1.

Abstract

The parameters that effect the synthesis of poly(styrene-co-divinylbenzene) magnetized with magnetite (STY-DVB-M) by polymerization emulsion were assessed in order to obtain magnetic beads to be used as matrix for lipase immobilization. The combined effect of polyvinyl alcohol (PVA) concentration and agitation was studied using response surface methodology. A 2 full-factorial design was employed for experimental design and analysis of the results. The optimum PVA concentration and agitation were found to be 1 wt% and 400 rpm, respectively. These conditions allow attaining the best particle size distribution of the synthesized particles (80% between 80 and 24 mesh). The performance of the magnetic beads was tested as a matrix for immobilizing two microbial lipases (Lipases from Burkholderia cepacia-BCL and Pseudomonas fluorescens-AKL) by physical adsorption and high immobilization yields (> 70%) and hydrolytic activities (≅ 1850 U g) were attained. The properties of free and immobilized lipases were searched and compared. Similar performance regarding the analyzed parameters (biochemical properties, kinetic constants and thermal stability) were obtained. Moreover, both immobilized lipases were found to be able to catalyze the transesterification of coconut oil with ethanol to produce fatty acid ethyl esters (FAEE). Further study showed that the B. cepacia immobilized lipase could be used seven times without significant decrease of activity, revealing half-life time of 970 h.

摘要

为了获得可用于脂肪酶固定化的基质的磁性珠,评估了通过聚合乳液合成与磁铁矿磁化的聚苯乙烯-二乙烯基苯(STY-DVB-M)的参数。使用响应面法研究了聚乙烯醇(PVA)浓度和搅拌的综合影响。采用 2 全因子设计进行实验设计和结果分析。发现最佳 PVA 浓度和搅拌速度分别为 1wt%和 400rpm。这些条件允许达到合成颗粒最佳的粒径分布(80%在 80 至 24 目之间)。测试了磁性珠作为固定化两种微生物脂肪酶(假单胞菌荧光酶 AKL 和洋葱伯克霍尔德菌脂肪酶 BCL)的基质的性能,通过物理吸附获得了高的固定化产率(>70%)和水解活性(≈1850U/g)。搜索和比较了游离和固定化脂肪酶的性质。在分析的参数(生化性质、动力学常数和热稳定性)方面获得了相似的性能。此外,两种固定化脂肪酶都能够催化椰子油与乙醇的酯交换反应生成脂肪酸乙酯(FAEE)。进一步的研究表明,固定化的洋葱伯克霍尔德菌脂肪酶可以重复使用七次而活性没有明显下降,半衰期为 970h。

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