Purification and Characterization of a Carboxylesterase from<i>Pseudomonas</i>sp. KWI-56

Sugihara, Akio; Shimada, Yuji; Nagao, Toshihiro; Iizumi, Taro; Nakamura, Kōichi; Fukase, Tetsuro; Tominaga, Yoshio

doi:10.1271/bbb.58.752

Cited by 13 publications

(3 citation statements)

References 6 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…DF20 Esterase 7 50 27.9 Özbek et al [102] Lactobacillus brevis NJ13 Esterase 8 50 48.12 Kim et al [103] Alcaligens faecalis Esterase 8 30 0.27 Poornima and Kasthuri [104] Burkholderia fungorum A216 Esterase 6.5 37 0.014 Jiao et al [105] Achromobacter denitrificans strain SP1 Esterase 8 50 89.5 Pradeep et al [106] Janthinobacterium lividum Esterase 7 30 0.00568 Park et al [107] Pseudomonas sp. KWI-56 Esterase 7.5 22 51.6 Sugihara et al [108] …”

Section: Resultsmentioning

confidence: 99%

Identification of lipolytic enzymes isolated from bacteria indigenous to Eucalyptus wood species for application in the pulping industry

Ramnath

Sitholé

Govinden

2017

Biotechnology Reports

103

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

Identification of lipolytic enzymes isolated from bacteria indigenous to Eucalyptus wood species for application in the pulping industry

Ramnath

Sitholé

Govinden

2017

Biotechnology Reports

103

View full text Add to dashboard Cite

show abstract

“…KWI-56 (Shimada et al, 1993;Sugihara et al, 1994). The construction of mutant plasmids coding mutant esterases is shown in Fig.…”

Section: Construction Of Plasmids To Express Mutant Esterasesmentioning

confidence: 99%

Hydrophobilization of an esterase by genetic combination with polyproline at the carboxyl terminal

Kwon

Ito

Ueda

et al. 1998

Biotechnol. Bioeng.

View full text Add to dashboard Cite

Polyproline, which is an amphiphilic polypeptide, was incorporated into the carboxyl terminal of an esterase by the recombinant DNA technique. The hydrophobicity of the esterase increased with increasing chain length of polyproline without inducing significant conformational changes. The mutant esterase catalyzed the hydrolysis of long‐chain carboxylic acid ester more efficiently than the native esterase. It is considered that the alteration of substrate specificity is due to enhanced access of the mutant esterases to hydrophobic substrates. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:582–586, 1998.

show abstract

“…(22,37,39,40). Esterase (EST) from Pseudomonas putida IFO12996 is capable of catalyzing the stereoselective hydrolysis of DL-MATI to produce DAT.…”

mentioning

confidence: 99%

Stereoselective Esterase from Pseudomonas putida IFO12996 Reveals α/β Hydrolase Folds for d -β-Acetylthioisobutyric Acid Synthesis

Elmi

Lee²,

Huang

et al. 2005

J Bacteriol

View full text Add to dashboard Cite

Esterase (EST) from Pseudomonas putida IFO12996 catalyzes the stereoselective hydrolysis of methyl DL-␤-acetylthioisobutyrate (DL-MATI) to produce D-␤-acetylthioisobutyric acid (DAT), serving as a key intermediate for the synthesis of angiotensin-converting enzyme inhibitors. The EST gene was cloned and expressed in Escherichia coli; the recombinant protein is a non-disulfide-linked homotrimer with a monomer molecular weight of 33,000 in both solution and crystalline states, indicating that these ESTs function as trimers. EST hydrolyzed DL-MATI to produce DAT with a degree of conversion of 49.5% and an enantiomeric excess value of 97.2% at an optimum pH of about 8 to 10 and an optimum temperature of about 57 to 67°C. The crystal structure of EST has been determined by X-ray diffraction to a resolution of 1.6 Å, confirming that EST is a member of the ␣/␤ hydrolase fold superfamily of enzymes and includes a catalytic triad of Ser97, Asp227, and His256. The active site is located approximately in the middle of the molecule at the end of a pocket ϳ12 Å deep. EST can hydrolyze the methyl ester group without affecting the acetylthiol ester moiety in DL-MATI. The examination of substrate specificity of EST toward other linear esters revealed that the enzyme showed specific activity toward methyl esters and that it recognized the configuration at C-2.Esterases belong to the hydrolase family and are ubiquitous enzymes identified in various species (24). These enzymes exist as either monomers or oligomers with subunit molecular weights (MW) that range from 25,000 to 60,000 (13), with a conserved pentapeptide (Gly-X 1 -Ser-X 2 -Gly) sequence around the catalytic serine residue (3), despite sharing a low sequence homology. Unlike lipases, esterases preferentially hydrolyze water-soluble esters or short-chain fatty acid triglycerides. The proposed mechanism of catalysis for esterases is similar to the serine protease mechanism, which involves nucleophilic attack by the catalytic serine hydroxyl on the carbonyl carbon of the scissile bond. This hydroxyl group becomes more nucleophilic and the reaction is stabilized through hydrogen bonding to the imidazole group of the catalytic histidine that is stabilized by the carboxyl group of the acidic member of the catalytic triad (9, 12). The importance of esterases for application in pharmaceutical and agrochemical industries is increasing because their properties as stereo-selective catalysts in the synthesis of optically pure molecules are diverse. On the basis of their protein structures, the ␣/␤ hydrolase architecture of lipases possesses an additional movable helical lid that controls access to the active site, providing a molecular explanation for lipase-enhanced activity upon contact with a lipase-water interface (41).D-␤-acetylthioisobutyric acid (DAT), also known as S-␤-acetylthio-2-methylpropionic acid, is a key intermediate for the synthesis of angiotensin-converting enzyme inhibitors, such as captopril and alacepril, which are used to treat hypertension and congestive he...

show abstract

Purification and Characterization of a Carboxylesterase fromPseudomonassp. KWI-56

Cited by 13 publications

References 6 publications

Identification of lipolytic enzymes isolated from bacteria indigenous to Eucalyptus wood species for application in the pulping industry

Identification of lipolytic enzymes isolated from bacteria indigenous to Eucalyptus wood species for application in the pulping industry

Hydrophobilization of an esterase by genetic combination with polyproline at the carboxyl terminal

Stereoselective Esterase from Pseudomonas putida IFO12996 Reveals α/β Hydrolase Folds for d -β-Acetylthioisobutyric Acid Synthesis

Contact Info

Product

Resources

About