Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12
Microbial proteases constitute one of the most important groups of industrially relevant enzymes. Proline iminopeptidases (PIPs) that specifically release amino-terminal proline from peptides are of major interest for applications in food biotechnology. Proline iminopeptidase has been extensively ch...
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ftunivmalaysia:oai:generic.eprints.org:101345 2023-11-12T04:08:25+01:00 Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12 Kamaruddin, Shazilah Ahmad Redzuan, Rohaiza Minor, Nurulermila Wan Seman, Wan Mohd. Khairulikhsan Md. Tab, Mahzan Jaafar, Nardiah Rizwana Ahmad Rodzli, Nazahiyah Jonet, Mohd. Anuar Bharudin, Izwan Yusof, Nur Athirah Quay, Doris Huai Xia 2022 application/pdf http://eprints.utm.my/101345/ http://eprints.utm.my/101345/1/NardiahRizwanaJaafar2022_BiochemicalCharacterisationandStructureDeterminationofaNovel.pdf https://doi.org/10.3390/catal12070722 en eng MDPI http://eprints.utm.my/101345/1/NardiahRizwanaJaafar2022_BiochemicalCharacterisationandStructureDeterminationofaNovel.pdf Kamaruddin, Shazilah and Ahmad Redzuan, Rohaiza and Minor, Nurulermila and Wan Seman, Wan Mohd. Khairulikhsan and Md. Tab, Mahzan and Jaafar, Nardiah Rizwana and Ahmad Rodzli, Nazahiyah and Jonet, Mohd. Anuar and Bharudin, Izwan and Yusof, Nur Athirah and Quay, Doris Huai Xia (2022) Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12. Catalysts, 12 (7). pp. 1-14. ISSN 2073-4344 TP Chemical technology Article PeerReviewed 2022 ftunivmalaysia https://doi.org/10.3390/catal12070722 2023-10-24T18:13:35Z Microbial proteases constitute one of the most important groups of industrially relevant enzymes. Proline iminopeptidases (PIPs) that specifically release amino-terminal proline from peptides are of major interest for applications in food biotechnology. Proline iminopeptidase has been extensively characterised in bacteria and filamentous fungi. However, no similar reports exist for yeasts. In this study, a protease gene from Glaciozyma antarctica designated as GaPIP was cloned and overexpressed in Escherichia coli. Sequence analyses of the gene revealed a 960 bp open reading frame encoding a 319 amino acid protein (35,406 Da). The purified recombinant GaPIP showed a specific activity of 3561 Umg-1 towards L-proline-p-nitroanilide, confirming its identity as a proline iminopeptidase. GaPIP is a cold-active enzyme with an optimum activity of 30? C at pH 7.0. The enzyme is stable between pH 7.0 and 8.0 and able to retain its activity at 10–30? C. Although GaPIP is a serine protease, only 25% inhibition by the serine protease inhibitor, phenylmethanesulfonylfluoride (PMSF) was recorded. This enzyme is strongly inhibited by the presence of EDTA, suggesting that it is a metalloenzyme. The dimeric structure of GaPIP was determined at a resolution of 2.4 Å. To date, GaPIP is the first characterised PIP from yeasts and the structure of GaPIP is the first structure for PIP from eukaryotes. Article in Journal/Newspaper Antarc* Antarctica Universiti Teknologi Malaysia: Institutional Repository Catalysts 12 7 722 |
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Universiti Teknologi Malaysia: Institutional Repository |
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TP Chemical technology |
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TP Chemical technology Kamaruddin, Shazilah Ahmad Redzuan, Rohaiza Minor, Nurulermila Wan Seman, Wan Mohd. Khairulikhsan Md. Tab, Mahzan Jaafar, Nardiah Rizwana Ahmad Rodzli, Nazahiyah Jonet, Mohd. Anuar Bharudin, Izwan Yusof, Nur Athirah Quay, Doris Huai Xia Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12 |
topic_facet |
TP Chemical technology |
description |
Microbial proteases constitute one of the most important groups of industrially relevant enzymes. Proline iminopeptidases (PIPs) that specifically release amino-terminal proline from peptides are of major interest for applications in food biotechnology. Proline iminopeptidase has been extensively characterised in bacteria and filamentous fungi. However, no similar reports exist for yeasts. In this study, a protease gene from Glaciozyma antarctica designated as GaPIP was cloned and overexpressed in Escherichia coli. Sequence analyses of the gene revealed a 960 bp open reading frame encoding a 319 amino acid protein (35,406 Da). The purified recombinant GaPIP showed a specific activity of 3561 Umg-1 towards L-proline-p-nitroanilide, confirming its identity as a proline iminopeptidase. GaPIP is a cold-active enzyme with an optimum activity of 30? C at pH 7.0. The enzyme is stable between pH 7.0 and 8.0 and able to retain its activity at 10–30? C. Although GaPIP is a serine protease, only 25% inhibition by the serine protease inhibitor, phenylmethanesulfonylfluoride (PMSF) was recorded. This enzyme is strongly inhibited by the presence of EDTA, suggesting that it is a metalloenzyme. The dimeric structure of GaPIP was determined at a resolution of 2.4 Å. To date, GaPIP is the first characterised PIP from yeasts and the structure of GaPIP is the first structure for PIP from eukaryotes. |
format |
Article in Journal/Newspaper |
author |
Kamaruddin, Shazilah Ahmad Redzuan, Rohaiza Minor, Nurulermila Wan Seman, Wan Mohd. Khairulikhsan Md. Tab, Mahzan Jaafar, Nardiah Rizwana Ahmad Rodzli, Nazahiyah Jonet, Mohd. Anuar Bharudin, Izwan Yusof, Nur Athirah Quay, Doris Huai Xia |
author_facet |
Kamaruddin, Shazilah Ahmad Redzuan, Rohaiza Minor, Nurulermila Wan Seman, Wan Mohd. Khairulikhsan Md. Tab, Mahzan Jaafar, Nardiah Rizwana Ahmad Rodzli, Nazahiyah Jonet, Mohd. Anuar Bharudin, Izwan Yusof, Nur Athirah Quay, Doris Huai Xia |
author_sort |
Kamaruddin, Shazilah |
title |
Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12 |
title_short |
Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12 |
title_full |
Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12 |
title_fullStr |
Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12 |
title_full_unstemmed |
Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12 |
title_sort |
biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, glaciozyma antarctica pi12 |
publisher |
MDPI |
publishDate |
2022 |
url |
http://eprints.utm.my/101345/ http://eprints.utm.my/101345/1/NardiahRizwanaJaafar2022_BiochemicalCharacterisationandStructureDeterminationofaNovel.pdf https://doi.org/10.3390/catal12070722 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
http://eprints.utm.my/101345/1/NardiahRizwanaJaafar2022_BiochemicalCharacterisationandStructureDeterminationofaNovel.pdf Kamaruddin, Shazilah and Ahmad Redzuan, Rohaiza and Minor, Nurulermila and Wan Seman, Wan Mohd. Khairulikhsan and Md. Tab, Mahzan and Jaafar, Nardiah Rizwana and Ahmad Rodzli, Nazahiyah and Jonet, Mohd. Anuar and Bharudin, Izwan and Yusof, Nur Athirah and Quay, Doris Huai Xia (2022) Biochemical characterisation and structure determination of a novel cold-active proline iminopeptidase from the psychrophilic yeast, Glaciozyma antarctica PI12. Catalysts, 12 (7). pp. 1-14. ISSN 2073-4344 |
op_doi |
https://doi.org/10.3390/catal12070722 |
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Catalysts |
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12 |
container_issue |
7 |
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722 |
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1782328714497884160 |