Gene cloning, sequencing, and expression of an esterase from Acinetobacter lwoffii I6C-1.

Abstract:

:The esterase-encoding gene, estA, was cloned from Acinetobacter lwoffii I6C-1 genomic DNA into Escherichia coli BL21(DE3) with plasmid vector pET-22b (pEM1). pEM1 has a 4.4-kb EcoRI insert that contained the complete estA gene. A 2.4-kb AvaI- SphI DNA fragment was subcloned (pEM3) and sequenced. estA gene encodes a protein of 366 amino acids (40,687 Da) with a pI of 9.17. The EstA signal peptide was 31 amino acids long, and the mature esterase sequence is 335 amino acids long (37.5 kDa). The conserved catalytic serine residue of EstA is in position 210. The EstA sequence was similar to that of the carboxylesterase from Acinetobacter calcoaceticus (75% identity, 85% similarity), Archaeoglobus fulgidus (37% identity, 59% similarity), and Mycobacterium tuberculosis (35% identity, 51% similarity). These enzymes contained the conserved motif G-X(1)-S-X(2)-G carrying the active-site serine of hydrolytic enzyme. The EstA activity in A. lwoffii I6C-1 remains constant throughout the stationary phase, and the activity in E. coil BL21 (DE3) with pEM1 was similar to A. lwoffii I6C-1.

journal_name

Curr Microbiol

journal_title

Current microbiology

authors

Kim HE,Lee IS,Kim JH,Hahn KW,Park UJ,Han HS,Park KR

doi

10.1007/s00284-002-3886-3

keywords:

subject

Has Abstract

pub_date

2003-04-01 00:00:00

pages

291-5

issue

4

eissn

0343-8651

issn

1432-0991

journal_volume

46

pub_type

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