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
Two novel esterase genes, <i>est2L</i> and <i>est4L</i>, 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...
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MDPI AG
2021-06-01
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| Series: | Applied Sciences |
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| author | Jong Eun Park Geum Seok Jeong Hyun Woo Lee Sung Kyum Kim Jungho Kim Hoon Kim |
| author_facet | Jong Eun Park Geum Seok Jeong Hyun Woo Lee Sung Kyum Kim Jungho Kim Hoon Kim |
| author_sort | Jong Eun Park |
| collection | DOAJ |
| description | Two novel esterase genes, <i>est2L</i> and <i>est4L</i>, 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 <i>p</i>-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 Co<sup>2+</sup> and Zn<sup>2+</sup>, 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. |
| first_indexed | 2024-03-10T10:03:19Z |
| format | Article |
| id | doaj.art-23ea1f3a4c3c4490916a74fb2ffeaaea |
| institution | Directory Open Access Journal |
| issn | 2076-3417 |
| language | English |
| last_indexed | 2024-03-10T10:03:19Z |
| publishDate | 2021-06-01 |
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| record_format | Article |
| series | Applied Sciences |
| spelling | doaj.art-23ea1f3a4c3c4490916a74fb2ffeaaea2023-11-22T01:42:50ZengMDPI AGApplied Sciences2076-34172021-06-011113590510.3390/app11135905Characterization 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 LibraryJong Eun Park0Geum Seok Jeong1Hyun Woo Lee2Sung Kyum Kim3Jungho Kim4Hoon Kim5Department of Pharmacy, and Research Institute of Life Pharmaceutical Sciences, Sunchon National University, Suncheon 57922, KoreaDepartment of Pharmacy, and Research Institute of Life Pharmaceutical Sciences, Sunchon National University, Suncheon 57922, KoreaDepartment of Pharmacy, and Research Institute of Life Pharmaceutical Sciences, Sunchon National University, Suncheon 57922, KoreaDepartment of Agricultural Chemistry, Sunchon National University, Suncheon 57922, KoreaDepartment of Agricultural Chemistry, Sunchon National University, Suncheon 57922, KoreaDepartment of Pharmacy, and Research Institute of Life Pharmaceutical Sciences, Sunchon National University, Suncheon 57922, KoreaTwo novel esterase genes, <i>est2L</i> and <i>est4L</i>, 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 <i>p</i>-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 Co<sup>2+</sup> and Zn<sup>2+</sup>, 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.https://www.mdpi.com/2076-3417/11/13/5905metagenomic libraryCczO-containing family IV esterasefamily V esterasesubstrate specificity |
| spellingShingle | Jong Eun Park Geum Seok Jeong Hyun Woo Lee Sung Kyum Kim Jungho Kim Hoon Kim 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 Applied Sciences metagenomic library CczO-containing family IV esterase family V esterase substrate specificity |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | metagenomic library CczO-containing family IV esterase family V esterase substrate specificity |
| url | https://www.mdpi.com/2076-3417/11/13/5905 |
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