scholarly journals Characterization of a Novel Family IV Esterase Containing a Predicted CzcO Domain and a Family V Esterase with Broad Substrate Specificity from an Oil-Polluted Mud Flat Metagenomic Library

2021 ◽  
Vol 11 (13) ◽  
pp. 5905
Author(s):  
Jong Eun Park ◽  
Geum Seok Jeong ◽  
Hyun Woo Lee ◽  
Sung Kyum Kim ◽  
Jungho Kim ◽  
...  
Keyword(s):  
Metagenomic Library ◽  
Signal Peptides ◽  
Mud Flat ◽  
Alkaline Conditions ◽  
Novel Esterase ◽  
Butyrylcholinesterase Activity ◽  
Triton X ◽  
Fusion Type ◽  
Family Iv

Two novel esterase genes, est2L and est4L, were identified from a previously constructed metagenomic library derived from an oil-polluted mud flat sample. The encoded Est2L and Est4L were composed of 839 and 267 amino acids, respectively, without signal peptides. Est2L was a unique fusion type of protein composed of two domains: a domain of the CzcO superfamily, associated with a cationic diffusion promoter with CzcD, and a domain of the acetylesterase superfamily, belonging to family IV with conserved motifs, such as HGG, GXSAG, and GXPP. Est2L was the first fused esterase with a CzcO domain. Est4L belonged to family V with GXS, GXSMGG, and PTL motifs. Native Est2L and Est4L were found to be in dimeric and tetrameric forms, respectively. Est2L and Est4L showed the highest activities at 60 °C and 50 °C, respectively, and at a pH of 10.0. Est2L preferred short length substrates, especially p-nitrophenyl (pNP)-acetate, with moderate butyrylcholinesterase activity, whereas Est4L showed the highest activity with pNP-decanoate and had broad specificity. Significant effects were not observed in Est2L from Co2+ and Zn2+, although Est2L contains the domain CzcD. Est2L and Est4L showed high stabilities in 30% methanol and 1% Triton X-100. These enzymes could be used for a variety of applications, such as detergent and mining processing under alkaline conditions.

scholarly journals Molecular Characterization of Novel Family IV and VIII Esterases from a Compost Metagenomic Library

2021 ◽  
Vol 9 (8) ◽  
pp. 1614
Author(s):  
Jong-Eun Park ◽  
Geum-Seok Jeong ◽  
Hyun-Woo Lee ◽  
Hoon Kim
Keyword(s):  
Metagenomic Library ◽  
Industrial Applications ◽  
Native Form ◽  
Metagenome Library ◽  
The Family ◽  
Alkaline Conditions ◽  
Molecular Masses ◽  
Novel Esterase ◽  
Family Iv

Two novel esterase genes, est8L and est13L, were isolated and identified from a compost metagenomic library. The encoded Est8L and Est13L had molecular masses of 33,181 and 44,913 Da consisting of 314 and 411 amino acids, respectively, without signal peptides. Est8L showed the highest identity (32.9%) to a hyper-thermophilic carboxylesterase AFEST from Archaeoglobus fulgidus compared to other esterases reported and was classified to be a novel member of family IV esterases with conserved regions such as HGGG, DY, GXSXG, DPL, and GXIH. Est13L showed the highest identity (98.5%) to the family VIII esterase Est7K from the metagenome library. Est8L and Est13L had the highest activities for p-nitrophenyl butyrate (C4) and p-nitrophenyl caproate (C6), respectively, and Est13L showed a broad substrate specificity for p-nitrophenyl substrates. Est8L and Est13L effectively hydrolyzed glyceryl tributyrate. The optimum temperatures for activities of Est8L and Est13L were identical (40 °C), and the optimum pH values were 9.0 and 10.0, respectively. Est13L showed higher thermostability than Est8L. Sephacryl S-200 HR chromatography showed that the native form of Est8L was a dimer. Interestingly, Est13L was found to be a tetramer, contrary to other family VIII esterases reported. Est8L was inhibited by 30% isopropanol, methanol, and acetonitrile; however, Est13L was activated to 182.9% and 356.1%, respectively, by 30% isopropanol and methanol. Est8L showed enantioselectivity for the S-form, but Est13L showed no enantioselectivity. These results show that intracellular Est8L and/or Est13L are oligomeric in terms of native forms and can be used for pharmaceutical and industrial applications with organic solvents under alkaline conditions.

scholarly journals Biochemical characterization of a family IV esterase with R-form enantioselectivity from a compost metagenomic library

2021 ◽  
Vol 64 (1) ◽  
Author(s):  
Jong Eun Park ◽  
Geum Seok Jeong ◽  
Hyun Woo Lee ◽  
Hoon Kim
Keyword(s):  
Biochemical Characterization ◽  
Metagenomic Library ◽  
Hormone Sensitive Lipase ◽  
Size Exclusion ◽  
Potential Candidate ◽  
Multiple Sequence ◽  
Exclusion Chromatography ◽  
Dimeric Form ◽  
Triton X ◽  
Family Iv

AbstractA novel family IV esterase (hormone-sensitive lipase, HSL) gene, est15L, was isolated from a compost metagenomic library. Encoded Est15L comprised 328 amino acids with a molecular weight of 34,770 kDa and was an intracellular esterase without a signal peptide. The multiple sequence alignment (MSA) of Est15L with other family IV esterases showed conserved regions such as HGG, DYR, GXSXG, DPL, and GXIH. Native Est15L was a dimeric form from the results of size exclusion chromatography. It was optimally active at 50 ℃ and pH 9.0, indicating alkaline esterase. However, it showed a low thermostability with half-lives of 30.3 at 30 ℃ and 2.7 min at 40 ℃. It preferred p-nitrophenyl butyrate (C4) with Km and Vmax values of 0.28 mM and 270.8 U/mg, respectively. Est15L was inhibited by organic solvents such as 30% methanol, isopropanol, and acetonitrile with residual activities of 12.5, 0.9, and 0.3%, respectively. It was also inhibited by 1% SDS and 1% PMSF; however, Est15L maintained its activity at 1% Triton X-100 and EDTA. Est15L was inhibited by Cu2+, Zn2+, Mn2+, Co2+, Fe2+, and Na+. In addition, Est15L hydrolyzed glyceryl tributyrate with a residual substrate amount of 43.7% at 60 min but could not hydrolyze the oils (fish and olive) and glyceryl trioleate. Interestingly, Est15L showed significant enantioselectivity toward the R-form with a residual substrate amount of 44.6%, lower than that of the S-form (83.5%). Considering its properties, Est15L can be a potential candidate for chemical reactions, such as the synthesis of pharmaceutical compounds.

Characterization of XtjR8: A novel esterase with phthalate-hydrolyzing activity from a metagenomic library of lotus pond sludge

2020 ◽  
Vol 164 ◽  
pp. 1510-1518
Author(s):  
Jiarong Qiu ◽  
Haiyan Yang ◽  
Zhenzhen Yan ◽  
Yaning Shi ◽  
Dandan Zou ◽  
...  
Keyword(s):  
Metagenomic Library ◽  
Novel Esterase

Characterization of Novel Family IV Esterase and Family I.3 Lipase from an Oil-Polluted Mud Flat Metagenome

2015 ◽  
Vol 57 (9) ◽  
pp. 781-792 ◽  
Author(s):  
Hee Jung Kim ◽  
Yu Seok Jeong ◽  
Won Kyeong Jung ◽  
Sung Kyum Kim ◽  
Hyun Woo Lee ◽  
...  
Keyword(s):  
Mud Flat ◽  
Family Iv

Screening and characterization of a novel esterase from a metagenomic library

2006 ◽  
Vol 45 (2) ◽  
pp. 315-323 ◽  
Author(s):  
Yun-Jung Kim ◽  
Gi-Sub Choi ◽  
Seung-Bum Kim ◽  
Gee-Sun Yoon ◽  
Yong-Sung Kim ◽  
...  
Keyword(s):  
Metagenomic Library ◽  
Novel Esterase

scholarly journals Identification and characterization of a novel carboxylesterase EstQ7 from a soil metagenomic library

2021 ◽  
Author(s):  
Zhenzhen Yan ◽  
Liping Ding ◽  
Dandan Zou ◽  
Luyao Wang ◽  
Yuzhi Tan ◽  
...  
Keyword(s):  
Metagenomic Library ◽  
Identification And Characterization

Characterization of a novel carboxylesterase with catalytic activity toward di(2-ethylhexyl) phthalate from a soil metagenomic library

2021 ◽  
Vol 785 ◽  
pp. 147260
Author(s):  
Zhenzhen Yan ◽  
Liping Ding ◽  
Dandan Zou ◽  
Jiarong Qiu ◽  
Yuting Shao ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Metagenomic Library ◽  
Ethylhexyl Phthalate

Preparative isolation and characterization of the B875 complex from Rhodobacter sphaeroides 2.4.1

1988 ◽  
Vol 66 (5) ◽  
pp. 442-448 ◽  
Author(s):  
Rafael Picorel ◽  
Gabriel Gingras
Keyword(s):  
Rhodobacter Sphaeroides ◽  
Phosphatidyl Ethanolamine ◽  
Sodium Dodecyl ◽  
Pigment Analysis ◽  
Preparative Isolation ◽  
Ethanolamine Phosphatidyl ◽  
Isolation And Characterization ◽  
Detergent System ◽  
Triton X

We have developed a simple and efficient method, using a mixed detergent system of sodium dodecyl sulfate and Triton X-100, for the preparative isolation of theB875 complex from Rhodobacter sphaeroides 2.4.1. As a bonus, the method allows the preparation of both the B875 and B800-850 complexes from the same batch of chromatophores. The preparations are spectrally pure, as indicated by absorption and circular dichroism spectroscopy. The latter method suggests that the Qy band of the B875 complex is due to weakly interacting bacteriochlorophyll molecules. Protein and pigment analysis shows that the B875 complex contains 2 mol of bacteriochlorophyll and 2 mol of sphaeroidene per mol of apoprotein (12 266 g), whereas the B800-850 complex contains 3 mol of bacteriochlorophyll and 1 mol of sphaeroidene per mol of apoprotein (11 497 g). While these stoichiometries are in accord with currently accepted models, they disagree with their published experimental basis. Phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol, and diphosphatidyl glycerol were found to be present in the B875 complex.

scholarly journals Identification and Characterization of a Halotolerant, Cold-Active Marine Endo-β-1,4-Glucanase by Using Functional Metagenomics of Seaweed-Associated Microbiota

2014 ◽  
Vol 80 (16) ◽  
pp. 4958-4967 ◽  
Author(s):  
Marjolaine Martin ◽  
Sophie Biver ◽  
Sébastien Steels ◽  
Tristan Barbeyron ◽  
Murielle Jam ◽  
...  
Keyword(s):  
Metagenomic Library ◽  
Functional Screening ◽  
Sample Collection ◽  
Prolonged Incubation ◽  
Content Type ◽  
Cold Active ◽  
Esterase Loci ◽  
Collection Period ◽  
Identification And Characterization

ABSTRACTA metagenomic library was constructed from microorganisms associated with the brown algaAscophyllum nodosum. Functional screening of this library revealed 13 novel putative esterase loci and two glycoside hydrolase loci. Sequence and gene cluster analysis showed the wide diversity of the identified enzymes and gave an idea of the microbial populations present during the sample collection period. Lastly, an endo-β-1,4-glucanase having less than 50% identity to sequences of known cellulases was purified and partially characterized, showing activity at low temperature and after prolonged incubation in concentrated salt solutions.

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