Identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading Bacillus sp. strain S3wahi
In this present study, characteristics and structure-function relationship of an organophosphate-degrading enzyme from Bacillus sp. S3wahi were described. S3wahi metallohydrolase, designated as S3wahi-MH (probable metallohydrolase YqjP), featured the conserved αβ/βα metallo-β-lactamase-fold (MBL-fol...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier B.V.
2024
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| Online Access: | http://psasir.upm.edu.my/id/eprint/116018/ http://psasir.upm.edu.my/id/eprint/116018/1/116018.pdf |
| _version_ | 1848866912773079040 |
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| author | Azman, Ameera Aisyah Muhd Noor, Noor Dina Leow, Adam Thean Chor Mohd Noor, Siti Aminah Mohamad Ali, Mohd Shukuri |
| author_facet | Azman, Ameera Aisyah Muhd Noor, Noor Dina Leow, Adam Thean Chor Mohd Noor, Siti Aminah Mohamad Ali, Mohd Shukuri |
| author_sort | Azman, Ameera Aisyah |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | In this present study, characteristics and structure-function relationship of an organophosphate-degrading enzyme from Bacillus sp. S3wahi were described. S3wahi metallohydrolase, designated as S3wahi-MH (probable metallohydrolase YqjP), featured the conserved αβ/βα metallo-β-lactamase-fold (MBL-fold) domain and a zinc bimetal at its catalytic site. The metal binding site of S3wahi-MH also preserves the H-X-H-X-D-H motif, consisting of specific amino acids at Zn1 (Asp69, His70, Asp182, and His230) and Zn2 (His65, His67, and His137). The multifunctionality of S3wahi-MH was demonstrated through a steady-state kinetic study, revealing its highest binding affinity (KM) and catalytic efficiency (kcat/KM) for OP compound, paraoxon, with values of 8.09 × 10−6 M and 4.94 × 105 M−1 s−1, respectively. Using OP compound, paraoxon, as S3wahi-MH native substrate, S3wahi-MH exhibited remarkable stability over a broad temperature range, 20 °C – 60 °C and a broad pH tolerance, pH 6–10. Corresponded to S3wahi-MH thermal stability characterization, the estimated melting temperature (Tm) was found to be 72.12 °C. S3wahi-MH was also characterized with optimum catalytic activity at 30 °C and pH 8. Additionally, the activity of purified S3wahi-MH was greatly enhanced in the presence of 1 mM and 5 mM of manganese (Mn2+), showing relative activities of 1323.68 % and 2073.68 %, respectively. The activity of S3wahi-MH was also enhanced in the presence of DMSO and DMF, showing relative activities of 270.37 % and 307.41 %, respectively. The purified S3wahi-MH retained >60 % residual activity after exposure to non-ionic Tween series surfactants. Nevertheless, the catalytic activity of S3wahi-MH was severely impacted by the treatment of SDS, even at low concentrations. Considering its enzymatic properties and promiscuity, S3wahi-MH emerges as a promising candidate as a bioremediation tool in wide industrial applications, including agriculture industry. |
| first_indexed | 2025-11-15T14:28:09Z |
| format | Article |
| id | upm-116018 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T14:28:09Z |
| publishDate | 2024 |
| publisher | Elsevier B.V. |
| recordtype | eprints |
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| spelling | upm-1160182025-03-28T00:59:18Z http://psasir.upm.edu.my/id/eprint/116018/ Identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading Bacillus sp. strain S3wahi Azman, Ameera Aisyah Muhd Noor, Noor Dina Leow, Adam Thean Chor Mohd Noor, Siti Aminah Mohamad Ali, Mohd Shukuri In this present study, characteristics and structure-function relationship of an organophosphate-degrading enzyme from Bacillus sp. S3wahi were described. S3wahi metallohydrolase, designated as S3wahi-MH (probable metallohydrolase YqjP), featured the conserved αβ/βα metallo-β-lactamase-fold (MBL-fold) domain and a zinc bimetal at its catalytic site. The metal binding site of S3wahi-MH also preserves the H-X-H-X-D-H motif, consisting of specific amino acids at Zn1 (Asp69, His70, Asp182, and His230) and Zn2 (His65, His67, and His137). The multifunctionality of S3wahi-MH was demonstrated through a steady-state kinetic study, revealing its highest binding affinity (KM) and catalytic efficiency (kcat/KM) for OP compound, paraoxon, with values of 8.09 × 10−6 M and 4.94 × 105 M−1 s−1, respectively. Using OP compound, paraoxon, as S3wahi-MH native substrate, S3wahi-MH exhibited remarkable stability over a broad temperature range, 20 °C – 60 °C and a broad pH tolerance, pH 6–10. Corresponded to S3wahi-MH thermal stability characterization, the estimated melting temperature (Tm) was found to be 72.12 °C. S3wahi-MH was also characterized with optimum catalytic activity at 30 °C and pH 8. Additionally, the activity of purified S3wahi-MH was greatly enhanced in the presence of 1 mM and 5 mM of manganese (Mn2+), showing relative activities of 1323.68 % and 2073.68 %, respectively. The activity of S3wahi-MH was also enhanced in the presence of DMSO and DMF, showing relative activities of 270.37 % and 307.41 %, respectively. The purified S3wahi-MH retained >60 % residual activity after exposure to non-ionic Tween series surfactants. Nevertheless, the catalytic activity of S3wahi-MH was severely impacted by the treatment of SDS, even at low concentrations. Considering its enzymatic properties and promiscuity, S3wahi-MH emerges as a promising candidate as a bioremediation tool in wide industrial applications, including agriculture industry. Elsevier B.V. 2024 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/116018/1/116018.pdf Azman, Ameera Aisyah and Muhd Noor, Noor Dina and Leow, Adam Thean Chor and Mohd Noor, Siti Aminah and Mohamad Ali, Mohd Shukuri (2024) Identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading Bacillus sp. strain S3wahi. International Journal of Biological Macromolecules, 271 (pt.1). art. no. 132395. pp. 1-19. ISSN 0141-8130; eISSN: 1879-0003 https://linkinghub.elsevier.com/retrieve/pii/S0141813024032008 10.1016/j.ijbiomac.2024.132395 |
| spellingShingle | Azman, Ameera Aisyah Muhd Noor, Noor Dina Leow, Adam Thean Chor Mohd Noor, Siti Aminah Mohamad Ali, Mohd Shukuri Identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading Bacillus sp. strain S3wahi |
| title | Identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading Bacillus sp. strain S3wahi |
| title_full | Identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading Bacillus sp. strain S3wahi |
| title_fullStr | Identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading Bacillus sp. strain S3wahi |
| title_full_unstemmed | Identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading Bacillus sp. strain S3wahi |
| title_short | Identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading Bacillus sp. strain S3wahi |
| title_sort | identification and characterization of a promiscuous metallohydrolase in metallo-β-lactamase superfamily from a locally isolated organophosphate-degrading bacillus sp. strain s3wahi |
| url | http://psasir.upm.edu.my/id/eprint/116018/ http://psasir.upm.edu.my/id/eprint/116018/ http://psasir.upm.edu.my/id/eprint/116018/ http://psasir.upm.edu.my/id/eprint/116018/1/116018.pdf |