%0 Journal Article
%T Molecular cloning and characterization of a N-acetylneuraminate lyase gene from Staphylococcus hominis<br>来源于葡萄球菌的N-乙酰神经氨酸裂合酶的基因克隆及性质
%A Chuanhua Zhou
%A Xi Chen
%A Jinhui Feng
%A Dongguang Xiao
%A Qiaqing Wu
%A Dunming Zhu
%A <br>周传华
%A 陈曦
%A 冯进辉
%A 肖冬光
%A 吴洽庆
%A 朱敦明
%J 生物工程学报
%D 2013
%I 
%X A N-acetylneuraminate lyase gene (shnal) from Staphylococcus hominis was cloned into pET-28a and expressed in Escherichia coli BL21 (DE3) host cells. The recombinant enzyme was purified and characterized. It is a homotetrameric enzyme with the optimum pH at 8.0 for the cleavage direction and the optimum pH and temperature were 7.5 and 45 °C for the synthetic direction. The activity of ShNAL is stable when incubated at 45 °C for 2 h but decreased rapidly over 50 °C. ShNAL showed high stability in a wide range pH from 5.0 to 10.0 with the residual activity being >70% when the enzyme was incubated in different buffers at 4 °C for 24 h. Its Km towards N-acetylneuraminic acid, pyruvate and ManNAc were (4.0±0.2) mmol/L, (35.1±3.2) mmol/L and (131.7±12.1) mmol/L, respectively. The kcat/Km value of Neu5Ac, ManNAc, and Pyr for ShNAL were 1.9 L/(mmol·s), 0.08 L/(mmol·s) and 0.08 L/(mmol·s), respectively.
%K Staphylococcus hominis
%K N-acetylneuraminate lyase
%K expression
%K purification
%K biocatalysis<br>葡萄球菌，N-乙酰神经氨酸裂合酶，表达，纯化，生物催化
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=F414F585DA9A500469956DB97CDE63A1&yid=FF7AA908D58E97FA&vid=771469D9D58C34FF&iid=E158A972A605785F&sid=03436AC72A659ACA&eid=F176272DD933A0BB&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=0