원문정보
초록
영어
A novel GDSLesterase from Sinorhizobium meliloti 1021 (EstSM), which has conserved amino acid sequences of Gly8, Asn9, Ser10, and Leu11 in N-terminal region, is characterized. Molecular weight of EstSM is 23kDa, and its pI is 4.89, respectively. EstSM has ~40% sequence identities with Mycobacterium smegmatis arylesterase(PDB ID: 2Q0Q) with putative catalytic sites of Ser10, Asp187, and His190. Using pQE30 vector, EstSM was expressed in Eschrichia coli
BL21 codon plus, and purified by His-tag affinity column. Activities of EstSM are investigated using various substrates such as p-nitrophenyl acetate and (R,S)-methyl-β-hydroxyisobutyrate. Biochemical and structural characterization of EstSM will provide the specificity regulation and activation mechanism of GDSL family in more detail. In addition, crystal studies for three dimensional structures are in progress.