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细菌肝素酶的特异性平板测定法。

Specific plate assay for bacterial heparinase.

作者信息

Zimmermann J J, Langer R, Cooney C L

机构信息

IBEX Technologies, Montreal, Quebec, Canada.

出版信息

Appl Environ Microbiol. 1990 Nov;56(11):3593-4. doi: 10.1128/aem.56.11.3593-3594.1990.

DOI:10.1128/aem.56.11.3593-3594.1990
PMID:2268165
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC185029/
Abstract

A procedure was developed for detecting heparinase activity on heparin agar plates. The method is based on the differential precipitation of heparin and heparinase-generated heparin fragments by protamine sulfate. Heparinase activity is detected by the presence of clear zones against a white background. This method can be used to screen for the expression of recombinant heparinase and to identify Flavobacterium heparinum mutants expressing heparinase constitutively.

摘要

开发了一种在肝素琼脂平板上检测肝素酶活性的方法。该方法基于硫酸鱼精蛋白对肝素和肝素酶产生的肝素片段的差异沉淀。通过白色背景上的透明区域来检测肝素酶活性。该方法可用于筛选重组肝素酶的表达，并鉴定组成型表达肝素酶的肝素黄杆菌突变体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fa9/185029/f400ac2f736d/aem00092-0372-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fa9/185029/9db75da9e473/aem00092-0371-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fa9/185029/f49ee37ffd2f/aem00092-0372-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fa9/185029/f400ac2f736d/aem00092-0372-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fa9/185029/9db75da9e473/aem00092-0371-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fa9/185029/f49ee37ffd2f/aem00092-0372-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fa9/185029/f400ac2f736d/aem00092-0372-b.jpg

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本文引用的文献

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Production of heparinase by Bacteroides.拟杆菌产生肝素酶。
J Bacteriol. 1961 Apr;81(4):595-604. doi: 10.1128/jb.81.4.595-604.1961.
2
Heparinase production by Flavobacterium heparinum.肝素黄杆菌产生肝素酶。
Appl Environ Microbiol. 1981 Feb;41(2):360-5. doi: 10.1128/aem.41.2.360-365.1981.
3
Differential anticoagulant activity of heparin fragments prepared using microbial heparinase.使用微生物肝素酶制备的肝素片段的抗凝活性差异
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Expression system for high levels of GAG lyase gene expression and study of the hepA upstream region in Flavobacterium heparinum.用于肝素黄杆菌中高水平GAG裂解酶基因表达的表达系统及甲型肝炎上游区域的研究。
J Bacteriol. 2002 Jun;184(12):3242-52. doi: 10.1128/JB.184.12.3242-3252.2002.
5
Isolation and expression in Escherichia coli of cslA and cslB, genes coding for the chondroitin sulfate-degrading enzymes chondroitinase AC and chondroitinase B, respectively, from Flavobacterium heparinum.分别编码来自肝素黄杆菌的硫酸软骨素降解酶软骨素酶AC和软骨素酶B的cslA和cslB基因在大肠杆菌中的分离与表达。
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Isolation and expression in Escherichia coli of hepB and hepC, genes coding for the glycosaminoglycan-degrading enzymes heparinase II and heparinase III, respectively, from Flavobacterium heparinum.从肝素黄杆菌中分别分离出编码糖胺聚糖降解酶肝素酶II和肝素酶III的乙肝和丙肝基因，并在大肠杆菌中进行表达。
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Purification, characterization and specificity of chondroitin lyases and glycuronidase from Flavobacterium heparinum.来自肝素黄杆菌的软骨素裂解酶和葡萄糖醛酸酶的纯化、特性及特异性
Biochem J. 1995 Dec 1;312 ( Pt 2)(Pt 2):569-77. doi: 10.1042/bj3120569.
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Flavobacterium heparinum 2-O-sulphatase for 2-O-sulphato-delta 4,5-glycuronate-terminated oligosaccharides from heparin.
Eur J Biochem. 1984 Dec 17;145(3):607-15. doi: 10.1111/j.1432-1033.1984.tb08600.x.
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Hydrolytic enzymes of anaerobic bacteria isolated from human infections.从人类感染中分离出的厌氧菌水解酶。
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Angiogenesis inhibition and tumor regression caused by heparin or a heparin fragment in the presence of cortisone.在可的松存在的情况下，肝素或肝素片段引起的血管生成抑制和肿瘤消退。
Science. 1983 Aug 19;221(4612):719-25. doi: 10.1126/science.6192498.
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Sequential degradation of heparin in Flavobacterium heparinum. Purification and properties of five enzymes involved in heparin degradation.肝素黄杆菌中肝素的顺序降解。参与肝素降解的五种酶的纯化及性质
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The enzymatic degradation of heparin and heparitin sulfate. I. The fractionation of a crude heparinase from flavobacteria.
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Flavobacterium heparinum 3-O-sulphatase for N-substituted glucosamine 3-O-sulphate.用于N-取代葡糖胺3-O-硫酸盐的肝素黄杆菌3-O-硫酸酯酶。
Eur J Biochem. 1985 Apr 15;148(2):359-65. doi: 10.1111/j.1432-1033.1985.tb08847.x.