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基于密度测定法的微分析测定聚羟基烷酸酯上脂肪酶解聚活性。

Densitometry based microassay for the determination of lipase depolymerizing activity on polyhydroxyalkanoate.

机构信息

Ecobiomaterial Research Laboratory, School of Biological Sciences, Universiti Sains Malaysia, Penang, 11800, Malaysia.

出版信息

AMB Express. 2013 May 8;3(1):22. doi: 10.1186/2191-0855-3-22.

DOI:10.1186/2191-0855-3-22
PMID:23657221
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3671206/
Abstract

A novel method for the assay of polyhydroxyalkanoate (PHA)-degrading ability of triacylglycerol lipases was developed. By applying the natural affinity of lipases towards hydrophobic interfaces, a sensitive and rapid densitometry analysis for the evaluation of hydrolytic activity of lipase droplets towards PHA-coated surface was successfully carried out. We found that 12 out of 14 tested lipases which are of fungal, bacterial and animal origin were able to hydrolyze P(3HB-co-92 mol% 4HB) thin film. The patterns and opacity of the hydrolysis spots of lipases on PHA films allowed easy comparison of PHA-hydrolytic strength of lipases. Lipase from the bacterium Chromobacterium viscosum exhibited the highest PHA-degrading activity. The hydrolytic activity of lipases on water insoluble PHA, emulsified p-nitrophenyl laurate and olive oil were also compared and interestingly some lipases showed better activity when PHA was used as a substrate.

摘要

开发了一种测定三酰基甘油脂肪酶聚羟基烷酸（PHA）降解能力的新方法。通过应用脂肪酶对疏水性界面的天然亲和力，成功地进行了灵敏快速的密度测定分析，以评估脂肪酶液滴对 PHA 涂层表面的水解活性。我们发现，在所测试的 14 种真菌、细菌和动物来源的脂肪酶中，有 12 种能够水解 P(3HB-co-92 mol%4HB)薄膜。脂肪酶在 PHA 薄膜上的水解斑点的形态和不透明度使得易于比较脂肪酶的 PHA 水解强度。来自粘质沙雷氏菌的脂肪酶表现出最高的 PHA 降解活性。还比较了脂肪酶在不溶于水的 PHA、乳化的对硝基苯棕榈酸酯和橄榄油上的水解活性，有趣的是，当 PHA 用作底物时，一些脂肪酶表现出更好的活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/48d603f9829b/2191-0855-3-22-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/635e3133a867/2191-0855-3-22-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/d1fb4bcfe186/2191-0855-3-22-2.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/647be477509e/2191-0855-3-22-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/f1aad7ae5c93/2191-0855-3-22-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/48d603f9829b/2191-0855-3-22-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/635e3133a867/2191-0855-3-22-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/d1fb4bcfe186/2191-0855-3-22-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/f11ecfff7d12/2191-0855-3-22-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/647be477509e/2191-0855-3-22-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/f1aad7ae5c93/2191-0855-3-22-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c02/3671206/48d603f9829b/2191-0855-3-22-6.jpg

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