Cloning, expression and characterization of a new 2-Cl-propionic acid ester hydrolase from B. subtilis

Glieder, Anton; Pressnig, Michaela; Schmidt, Andrea; Stanzer, Thomas; Reisner, Andreas; Schwab, Helmut

doi:10.1016/s1381-1177(02)00173-x

Cited by 7 publications

(6 citation statements)

References 28 publications

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“…Successful overexpression of EstBL was achieved using the pMS470 ⌬ 8 vector. The pMS470 ⌬ 8 vector was first described by Balzer et al [1992] and has been used successfully to express a number of esterase encoding genes from the genera Burkholderia [Reiter et al, 2000;Petersen et al, 2001] and Bacillus [Glieder et al, 2002]. The success of this expression vector has been attributed to a combination of the Shine-Dalgarno sequence of T7 gene 10 and the weaker tac -promoter [Balzer et al, 1992].…”

Section: Discussionmentioning

confidence: 99%

Molecular Characterization of a Novel Family VIII Esterase from <i>Burkholderia multivorans</i> UWC10

Rashamuse¹,

Burton

Stafford

et al. 2007

Microb Physiol

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An esterase producing Burkholderia multivorans UWC10 strain was isolated by culture enrichment. A shotgun library of B. multivorans UWC10 genomic DNA was screened for esterase activity and a recombinant clone conferring an esterolytic phenotype was identified. Full-length sequencing of the DNA insert showed that it consisted of a single open reading frame (ORF1) encoding a predicted protein of 398 amino acids. ORF1 (termed EstBL) had a high protein sequence identity to family VIII esterases. The EstBL primary structure showed two putative serine motifs, G-V-S₁₄₉-D-G and S₇₄-V-T-K. The estBL gene was successfully over-expressed in E. coli and the encoded protein purified by a combination of ammonium sulphate fractionation, hydrophobic interaction, ion exchange and size exclusion chromatographies. Biochemical assays confirmed EstBL esterase activity and revealed a preference for short-chain p-nitrophenyl and β-naphthyl esters (C2-C4) with no activity against β-lactam substrates. Secondary structure predictions indicated that EstBL adopts the α/β fold, which is common to all esterases.

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Section: Discussionmentioning

confidence: 99%

Molecular Characterization of a Novel Family VIII Esterase from <i>Burkholderia multivorans</i> UWC10

Rashamuse¹,

Burton

Stafford

et al. 2007

Microb Physiol

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“…In a similar study Glieder et al (2002), while isolating and expressing a 2-CL-propionic acid ester hydrolase from genomic library of B. subtilis , found three ORFs in the cloned fragment that showed esterase activity. The B. subtilis insert carried two truncated ORFs and one complete ORF that encodes an esterase in the clone.…”

Section: Resultsmentioning

confidence: 84%

“…The B. subtilis insert carried two truncated ORFs and one complete ORF that encodes an esterase in the clone. Glieder et al (2002) cloned the esterase gene (Est4B) from Bacillus subtilis , which was confirmed on tributyrin plates. The protein sequence revealed high homology to the product of the srfD gene, a putative thioesterase gene from the surfactin synthetase gene cluster (Glieder et al 2002).…”

Section: Resultsmentioning

confidence: 98%

“…Glieder et al (2002) cloned the esterase gene (Est4B) from Bacillus subtilis , which was confirmed on tributyrin plates. The protein sequence revealed high homology to the product of the srfD gene, a putative thioesterase gene from the surfactin synthetase gene cluster (Glieder et al 2002). They reported that ORF 2 in the insert encodes a 243-amino acid est4B1 with 99 % sequence identity at the amino acid level to thioesterases.…”

Section: Resultsmentioning

confidence: 98%

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Isolation of a thioesterase gene from the metagenome of a mountain peak, Apharwat, in the northwestern Himalayas

Sudan

Vakhlu

2012

3 Biotech

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“…Acinetobacter [8,9], Bacillus [10,11] and Pseudomonas [12,13] are examples from various microbial strains that possess identifing and cloning of esterases and lipases. The studies have been reported that, metal ions and organic solvent resistance related to microbial esterases and lipases [14].…”

Section: Lipase and Esterase Definitionmentioning

confidence: 99%

Lipases, Definition, and their Application

Moayd¹,

Yan²

2017

IOSRJPBS

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Esterase and lipase have been consisted of α/β hydroxylase super family that characterized primarily by their α/β hydroxylase fold (common fold), which is in the center, pre-dominantly parallel β-sheet and flanked via the connections of α-helical. Nucleophilic elbow is an esterases and lipases that shared the characteristic sequence motifs (GXSXG I. Lipase and Esterase DefinitionEstarase and lipase are cosisted of α/β hydroxylase super famliy that characterized primarily by their α/β hydroxylase fold (common fold), which is in the center, pre-dominantly parallel β-sheet and flanked via the connections of α-helical. Nucleophilic elbow is esterases and lipases that shared the characteristic sequence motifs (GXSXG) [1,2]. This motif is embeded by serine, and the mediation of ester hydrolysis by attacking the nucleophilic of serine active on the substrate carbonyl in the system of a charge relay with other two residues of amino acid (histidine and aspartic acid) [1]. The residues of these amino acids compose a triad of catalyse in the specific-order (serine -aspartic acid -histidine) in the chain of polypeptide [3][4][5][6]. Forthat, the most of lipases and esterases resemble serine-proteases and lipases in hydrolytic mechanims. Esterases (EC 3.1.1.1), as well as lipases (EC 3.1.1.3)are the versatile classes of biocatalysts due to their region-, enatio-selectivity and high specificity. Esterases are those that can catalyze the hydrolysis of less than 10 short-acyl-chain-lengthed triglycerides, whereas, lipases are catalysts that can hydrolyze more than 10 long-acyl-chain-lengthed triglycerides. Studies of the structure displayed that the presene or absence of lid structure caused the diffrentiation between esterases and lipases and responsible for lipase interfacial activation [4][5][6][7]. Life kindoms has a widely distribution from esterase and lipases.In a widly ranging from organisms, esterases and lipases have been identified and cloned severel of them. Acinetobacter [8,9], Bacillus [10,11] and Pseudomonas [12,13] are examples from various microbial strains that possess identifing and cloning of esterases and lipases. The studies have been reported that, metal ions and organic solvent resistance related to microbial esterases and lipases [14]. Thus, they should be catalysis effeceintly in organic synthisis for an enantio-and regionselective reactions, among the groups biocatalysis that have been the most important in biotechnology. The optically pure substances in organic synthesis have been used esterases and lipases successfully. II. Lipase applicationLipases applications can be widely produced from a bacterial and fungal production. In fact, they have stability in organic solvent wide range and many present high thermostabilities, they have optimal range of temperature and wider pH than eukaryotic origin lipases, and have a high diverse range of substrate with high enantio-and regioselectivity making an important microbial lipases and attractive choice in organic synthesis for many applications. Li...

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Cloning, expression and characterization of a new 2-Cl-propionic acid ester hydrolase from B. subtilis

Cited by 7 publications

References 28 publications

Molecular Characterization of a Novel Family VIII Esterase from <i>Burkholderia multivorans</i> UWC10

Molecular Characterization of a Novel Family VIII Esterase from <i>Burkholderia multivorans</i> UWC10

Isolation of a thioesterase gene from the metagenome of a mountain peak, Apharwat, in the northwestern Himalayas

Lipases, Definition, and their Application

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