4-Oxalocrotonate tautomerase,酶由62氨基酸残基/单体。
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陈韩,肯扬GL,科廷F, Harayama年代,Bembenek我,Hajipour G,惠特曼CP
4-Oxalocrotonate tautomerase,酶由62氨基酸残基/单体。
生物化学杂志。1992年9月5日;267(25):17716 - 21所示。
- PubMed ID
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1339435 (在PubMed]
- 文摘
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xylH基因编码4-oxalocrotonate tautomerase (4-OT)位于假单胞菌的subclone putida mt-2 TOL质粒pWW0并插入一个大肠杆菌表达载体。几个metafission通路的基因编码的pWW0在大肠杆菌克隆,但他们的基因产物的过度会见了有限的成功。利用大肠杆菌碱性磷酸酶启动子(phoA)加上ribosome-binding区域的正确定位,我们能够表达功能4-OT收益率至少10毫克纯酶/升的文化。4-OT以前描述和证明是一个极其有效的催化剂(惠特曼,c·P。、Aird b。Gillespie, w·R。Stolowich, n . j . (1991) j。化学。Soc。113年,3154 - 3162)。动能和大肠coli-expressed蛋白质的物理特征表明,它是相同的与从p . putida 4-OT孤立。的功能单元是一个五聚物相同的子单元,每个只有62个氨基酸残基组成。 This is the smallest enzyme subunit reported to date. The amino acid sequence, determined in part from automated Edman degradation and also deduced from the primary sequence of xylH, did not show homology with any of the sequences in the current data bases nor with any of the sequences of enzymes that catalyze similar reactions. We propose that the active site of 4-OT may be established by an overlap of subunits and comprised of amino acid residues belonging to several, if not all, of the subunits.