Fingerprint lipolytic enzymes with chromogenic p-nitrophenyl esters of structurally diverse carboxylic acids

Qian, Le; Liu, Jia-Yan; Liu, Jiaying; Yu, Hui‐Lei; Li, Chun-Xiu; Xu, Jian‐He

doi:10.1016/j.molcatb.2011.07.010

Cited by 8 publications

(10 citation statements)

References 24 publications

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“…Kanamycin (50 μg ml Table S1. Cells were cultured as described previously (Qian et al 2011). Vector pET-28a (+) for heterogeneous expression studies (Chen et al 2015) was obtained from Novagen (Shanghai, China).…”

Section: Bacterial Strains and Vectorsmentioning

confidence: 99%

Identification of a new thermostable and alkali-tolerant α-carbonic anhydrase from Lactobacillus delbrueckii as a biocatalyst for CO2 biomineralization

Jiang

Yongjuan

et al. 2015

Bioresour. Bioprocess.

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Background: Carbonic anhydrase (CA, EC 4.2.1.1), an ancient enzyme and the fastest among many enzymes, is a useful biocatalyst for carbon capture use and storage (CCUS). The use of alkaline buffers and high temperatures are favorable for biomineralization. Hence, the stability of CA under such harsh conditions is extremely important for its practical application. Methods and results:Herein, we report a new thermostable and alkaline-tolerant α-CA (designated as LdCA), with only 26 % identity to bovine CA (BCA), which was identified by genome mining from Lactobacillus delbrueckii CGMCC 8137. It was overexpressed in Escherichia coli in a soluble form and purified to electrophoretic homogeneity by HisTrap affinity chromatography. The dimer protein had a subunit molecular weight of 23.8 kDa and showed extremely high stability at pH 6.0-11.0 and 30-60 °C. Its activity was maintained even after incubation at 90 °C for 15 min. The half-lives of the enzyme measured at 30, 40, and 50 °C were 630, 370, and 177 h, respectively. At pH 9.0, 10.0, and 11.0, its half-lives were 105, 65, and 41 min, respectively. LdCA was applied at 50 °C to accelerate the formation of calcium carbonate in a vaterite phase. Conclusions:In summary, a new CA with high thermal and alkaline stability was identified from a general bacterium, demonstrating an effective strategy for discovering new and useful biocatalysts.

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Section: Bacterial Strains and Vectorsmentioning

confidence: 99%

Identification of a new thermostable and alkali-tolerant α-carbonic anhydrase from Lactobacillus delbrueckii as a biocatalyst for CO2 biomineralization

Jiang

Yongjuan

et al. 2015

Bioresour. Bioprocess.

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“…RhEst1 did not display any hydrolysis activity toward benzoyl ester (S 18 ). Compared with other esterases reported previously, RhEst1 has a relatively high activity toward pNP propionate (39).…”

Section: Resultsmentioning

confidence: 55%

“…Eighteen chromogenic pNP esters with different acyl groups (Fig. 5), including aliphatic, cyclic, and aromatic acyls, were chosen from the chromogenic pNP ester library developed recently in our laboratory for rapid fingerprinting of lipolytic/esterolytic enzymes (39).…”

Section: Resultsmentioning

confidence: 99%

Cloning and Characterization of a Novel Esterase from Rhodococcus sp. for Highly Enantioselective Synthesis of a Chiral Cilastatin Precursor

Zhang

Pan

Luan

et al. 2014

Appl Environ Microbiol

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bA novel nonheme chloroperoxidase (RhEst1), with promiscuous esterase activity for enantioselective hydrolysis of ethyl (S)-2,2-dimethylcyclopropanecarboxylate, was identified from a shotgun library of Rhodococcus sp. strain ECU1013. RhEst1 was overexpressed in Escherichia coli BL21(DE3), purified to homogeneity, and functionally characterized. Fingerprinting analysis revealed that RhEst1 prefers para-nitrophenyl (pNP) esters of short-chain acyl groups. pNP esters with a cyclic acyl moiety, especially that with a cyclobutanyl group, were also substrates for RhEst1. The K m values for methyl 2,2-dimethylcyclopropanecarboxylate (DmCpCm) and ethyl 2,2-dimethylcyclopropane carboxylate (DmCpCe) were 0.25 and 0.43 mM, respectively. RhEst1 could serve as an efficient hydrolase for the bioproduction of optically pure (S)-2,2-dimethyl cyclopropane carboxylic acid (DmCpCa), which is an important chiral building block for cilastatin. As much as 0.5 M DmCpCe was enantioselectively hydrolyzed into (S)-DmCpCa, with a molar yield of 47.8% and an enantiomeric excess (ee) of 97.5%, indicating an extremely high enantioselectivity (E ‫؍‬ 240) of this novel and unique biocatalyst for green manufacturing of highly valuable chiral chemicals.

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“…Glycerin tributyrate and Tween-80 were purchased from TCI (Tokyo, Japan). dl-Menthyl benzoate, dl-menthyl acetate, dl-menthyl butyrate, dl-menthyl chloroacetate, and p-nitrophenyl butyrate (pNPB) were synthesized in our laboratory [9,16]. The other chemicals used in this work were of analytical grade from local sources.…”

Section: Methodsmentioning

confidence: 99%

Cloning and Characterization of an Enantioselective l-Menthyl Benzoate Hydrolase from Acinetobacter sp. ECU2040

Yin

Zheng

et al. 2015

Appl Biochem Biotechnol

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A new esterase gene abmbh, encoding a benzoate hydrolase which can enantioselectively hydrolyze l-menthyl benzoate to l-menthol, was recently identified from the genomic library of a soil isolate Acinetobacter sp. ECU2040. The abmbh gene contains a 1080-bp open reading frame encoding a protein of 360 amino acids with a calculated molecular mass of 40.7 kDa. The corresponding enzyme AbMBH was functionally expressed in Escherichia coli BL21 (DE3), purified, and characterized. The AbMBH displayed the maximum activity towards p-nitrophenyl butyrate at 50 °C, and an optimum pH of 8.5. A K M of 2.6 mM and a k cat of 0.26 s(-1) were observed towards dl-menthyl benzoate. The AbMBH exhibited a moderate enantioselectivity (E = 27.5) towards dl-menthyl benzoate. It can also catalyze the enantioselective hydrolysis of a variety of racemic menthyl esters, including dl-menthyl acetate, dl-menthyl chloroacetate, and dl-menthyl butyrate.

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Fingerprint lipolytic enzymes with chromogenic p-nitrophenyl esters of structurally diverse carboxylic acids

Cited by 8 publications

References 24 publications

Identification of a new thermostable and alkali-tolerant α-carbonic anhydrase from Lactobacillus delbrueckii as a biocatalyst for CO2 biomineralization

Identification of a new thermostable and alkali-tolerant α-carbonic anhydrase from Lactobacillus delbrueckii as a biocatalyst for CO2 biomineralization

Cloning and Characterization of a Novel Esterase from Rhodococcus sp. for Highly Enantioselective Synthesis of a Chiral Cilastatin Precursor

Cloning and Characterization of an Enantioselective l-Menthyl Benzoate Hydrolase from Acinetobacter sp. ECU2040

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