1994
DOI: 10.1016/0378-1119(94)90630-0
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The Bacillus subtilis pnbA gene encoding p-nitrobenzyl esterase: cloning, sequence and high-level expression in Escherichia coli

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Cited by 86 publications
(57 citation statements)
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“…The 1.8 kb product was cut from the gel and the DNA extracted using a Qiaex kit. The ®nal products were ligated with the vector generated by BamHI-XbaI digestion of pNB106R (Zock et al, 1994). This library was used to transform competent E. coli TG1 cells, as described (Moore & Arnold, 1996).…”
Section: Dna Shufflingmentioning
confidence: 99%
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“…The 1.8 kb product was cut from the gel and the DNA extracted using a Qiaex kit. The ®nal products were ligated with the vector generated by BamHI-XbaI digestion of pNB106R (Zock et al, 1994). This library was used to transform competent E. coli TG1 cells, as described (Moore & Arnold, 1996).…”
Section: Dna Shufflingmentioning
confidence: 99%
“…Bacillus subtilis p-nitrobenzyl (pNB) esterase catalyzes the hydrolysis of the para-nitrobenzyl esters of various cephalosporin-type antibiotics, a necessary step in their large-scale synthesis (Zock et al, 1994). Using four generations of sequential random mutagenesis and screening, we evolved a series of pNB esterases up to 30 times more active towards hydrolysis of the pNB ester of loracarbef (LCNpNB) in aqueous dimethylformamide (Moore & Arnold, 1996).…”
Section: Introductionmentioning
confidence: 99%
“…( Table 1) as those shown among the microbial lipolytic enzymes, LipA, LipB, YtpA, and PnbA of B. subtilis, and PldB and Aes of E. coli [8][9][10][11][12][13][14] . This suggests that YkoN has a lipolytic activity, though overall homology of YkoN with those of the lipase/esterase is not significant.…”
Section: Resultsmentioning
confidence: 70%
“…The results suggested that ykoN is not involved in the glucosylated diacylglycerides metabolismbut that it is involved in other function. Deep inspection of the amino acid sequence of YkoN revealed a pentapeptide sequence (Gly-Gly-Ser-Met-Gly) that is compatible with the Gly/Ala-X-Ser-X-Gly motif of lipase/esterase flanked with four hydrophobic residues (Ile-Ile-Ile-Thr) on the N-terminal side of the motif Amino acid sequences aligned are YkoN (BSORF), LipA (BSORF), LipB 10) , YtpA 11) and PnbA/EstB 12) from B. subtilis, and PldB 13) and Aes 14) from E. coli.…”
Section: Resultsmentioning
confidence: 83%
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