Expression of a novel hyaluronidase from Streptococcus zooepidemicus in Escherichia coli and its application for the preparation of HA oligosaccharides

Xueping Guo; Fei Liu; Xiqiang Zhu

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Expression of a novel hyaluronidase from Streptococcus zooepidemicus in Escherichia coli and its application for the preparation of HA oligosaccharides

机译：一种新型的兽疫链球菌透明质酸酶在大肠杆菌中的表达及其在制备HA寡糖中的应用

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Hyaluronan (HA) oligosaccharides which can stimulate angiogenesis and suppress the growth of tumors have attracted more and more attention. In order to prepare pure and well-defined oligosaccharides from high-molecular-weight HA in a rapid and simple manner, an enzymatic degradation method was developed, which included degradation with a novel recombinant hyaluronan lyase (HA lyase, hyaluronidase, or HAase) and gel permeation chromatography. The HAase protein was expressed in Escherichia coli with the expression vector pBV220. The HAase was purified and refolded, and specific activity of the enzyme solution was 3800 U/mg. HA was degraded with HAase at the optimized conditions, yielding 46% and 31% of HA disaccharides and HA tetrasaccharides, respectively. These HA oligosaccharides were conveniently separated by consecutive column chromatography on Bio-gel P6 and were identified by HPLC-MS.

机译：能够刺激血管生成并抑制肿瘤生长的透明质酸（HA）寡糖已引起越来越多的关注。为了以快速，简便的方式从高分子量HA制备纯净和定义明确的寡糖，开发了一种酶促降解方法，包括用新型重组透明质酸裂解酶（HA裂解酶，透明质酸酶或HAase）降解，以及凝胶渗透色谱。 HAase蛋白用表达载体pBV220在大肠杆菌中表达。纯化并重新折叠HAase，酶溶液的比活为3800 U / mg。 HA在最适条件下用HAase降解，分别产生46％和31％的HA二糖和HA四糖。这些HA低聚糖可通过Bio-gel P6上的连续柱色谱法方便地分离，并通过HPLC-MS鉴定。

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《Carbohydrate Polymers: Scientific and Technological Aspects of Industrially Important Polysaccharides》 |2009年第2期|共7页
作者
Xueping Guo; Fei Liu; Xiqiang Zhu;
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正文语种 eng
中图分类天然化合物;
关键词
Hyaluronan oligosaccharides; Enzymatic hydrolysis; Hyaluronan lyase; Streptococcus zooepidemicus; Cloning and expression;

机译：透明质酸寡糖;酶水解;透明质酸裂合酶;兽疫链球菌;克隆与表达;

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1. Expression of a novel hyaluronidase from Streptococcus zooepidemicus in Escherichia coli and its application for the preparation of HA oligosaccharides [J] . Xueping Guo, Fei Liu, Xiqiang Zhu Carbohydrate Polymers: Scientific and Technological Aspects of Industrially Important Polysaccharides . 2009,第2期

机译：一种新型的兽疫链球菌透明质酸酶在大肠杆菌中的表达及其在制备HA寡糖中的应用
2. Exclusion of bioactive contaminations in Streptococcus pyogenes erythrogenic toxin A preparations by recombinant expression in Escherichia coli. [J] . U Fagin, U Hahn, J Gr?tzinger, Infection and immunity . 1997,第11期

机译：通过在大肠杆菌中的重组表达排除化脓性链球菌促红细胞生成素A制剂中的生物活性污染物。
3. Human N-Acetylglucosaminyltransferase I. Expression in Escherichia coli as a Soluble Enzyme, and Application as an Immobilized Enzyme for the Chemoenzymatic Synthesis of N-Linked Oligosaccharides [J] . Kazuhito Fujiyama, Yoshihiro Ido, Ryo Misaki, Journal of Bioscience and Bioengineering . 2001,第6期

机译：人N-乙酰氨基葡萄糖氨基转移酶I.在大肠杆菌中的表达作为一种可溶性酶，以及作为固定化酶用于化学合成N-连接寡糖的固定化酶
4. Optimization of Recombinant Bacteriophage Lysin of Streptococcus Expression in Escherichia coli [C] . Wang Pu, Wang Xiaofeng, Chen Weiqing 2010 4th International Conference on Bioinformatics and Biomedical Engineering . 2010

机译：重组链球菌噬菌体溶菌素在大肠杆菌中表达的优化
5. Effect of prebiotic oligosaccharides on enteropathogenic Escherichia coli adherence. [D] . Shoaf, Kari. 2007

机译：益生元寡糖对肠致病性大肠杆菌粘附的影响。
6. Sequence analysis and expression in Escherichia coli of the hyaluronidase gene of Streptococcus pyogenes bacteriophage H4489A. [O] . W L Hynes, J J Ferretti 1989

机译：化脓性链球菌噬菌体H4489A的透明质酸酶基因的序列分析和在大肠杆菌中的表达。
7. Exclusion of bioactive contaminations in Streptococcus pyogenes erythrogenic toxin A preparations by recombinant expression in Escherichia coli [O] . U Fagin, U Hahn, J Grötzinger, 1997

机译：在大肠杆菌中的重组表达中，在细胞内毒素中排除生物活性污染物的红细胞毒素A制剂

Expression of a novel hyaluronidase from Streptococcus zooepidemicus in Escherichia coli and its application for the preparation of HA oligosaccharides

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