NUCLEOTIDE-SEQUENCE OF A NOVEL ARYLESTERASE GENE FROM VIBRIO-MIMICUS AND CHARACTERIZATION OF THE ENZYME EXPRESSED IN ESCHERICHIA-COLI

作者：SHAW JF^*; CHANG RC; CHUANG KH; YEN YT; WANG YJ; WANG FG

来源：Biochemical Journal, 1994, 298: 675-680.

A gene coding for an arylesterase of Vibrio mimicus was cloned. Sequence determination reveals that the esterase gene has an open reading frame of 600 nucleotides which encodes a protein of M(r) 22 300. The deduced amino acid sequence contains a pentapeptide GDSLS (residues 27-31), which was also found in the phospholipid-cholesterol acyltransferase from Aeromonas hydrophila. Substitution of Ser-29 by alanine or cysteine in the cloned gene abolished the esterase activity in the tributyrin plate assay. On the other hand, the activity was not lost when Ser-31 was changed to alanine. The cloned gene was expressed in Escherichia coli, and the protein purified by a four-step procedure. The purified protein migrated on SDS/PAGE as a single band with an apparent M(r) of 22 100. This enzyme favoured the hydrolysis of several arylesters and was classified as an arylesterase (EC 3.1.1.2). N-Terminal analysis showed that Ser-20 was the first amino acid of the mature secreted protein, suggesting that the N-terminal 19 hydrophobic amino acids served as a signal peptide.

• 出版日期1994-3-15
• 单位中国科学院植物研究所