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Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA

Xylanases (EC 3.2.1.8) are essential enzymes due to their applications in various industries such as textile, animal feed, paper and pulp, and biofuel industries. Halo-thermophilic Rhodothermaceae bacterium RA was previously isolated from a hot spring in Malaysia. Genomic analysis revealed that this...

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Main Authors: Liew, Kok Jun, Ngooi, Chen Yi, Shamsir, Mohd. Shahir, Sani, Rajesh Kumar, Chong, Chun Shiong, Goh, Kian Mau
Format: Article
Published: Academic Press Inc. 2019
Subjects:
QD Chemistry
Online Access:http://eprints.utm.my/id/eprint/89065/
http://dx.doi.org/10.1016/j.pep.2019.105464
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spelling my.utm.890652021-01-26T08:42:03Z http://eprints.utm.my/id/eprint/89065/ Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA Liew, Kok Jun Ngooi, Chen Yi Shamsir, Mohd. Shahir Sani, Rajesh Kumar Chong, Chun Shiong Goh, Kian Mau QD Chemistry Xylanases (EC 3.2.1.8) are essential enzymes due to their applications in various industries such as textile, animal feed, paper and pulp, and biofuel industries. Halo-thermophilic Rhodothermaceae bacterium RA was previously isolated from a hot spring in Malaysia. Genomic analysis revealed that this bacterium is likely to be a new genus of the family Rhodothermaceae. In this study, a xylanase gene (1140 bp) that encoded 379 amino acids from the bacterium was cloned and expressed in Escherichia coli BL21(DE3). Based on InterProScan, this enzyme XynRA1 contained a GH10 domain and a signal peptide sequence. XynRA1 shared low similarity with the currently known xylanases (the closest is 57.2–65.4% to Gemmatimonadetes spp.). The purified XynRA1 achieved maximum activity at pH 8 and 60 °C. The protein molecular weight was 43.1 kDa XynRA1 exhibited an activity half-life (t1/2) of 1 h at 60 °C and remained stable at 50 °C throughout the experiment. However, it was NaCl intolerant, and various types of salt reduced the activity. This enzyme effectively hydrolyzed xylan (beechwood, oat spelt, and Palmaria palmata) and xylodextrin (xylotriose, xylotetraose, xylopentaose, and xylohexaose) to produce predominantly xylobiose. This xylanase is the first functionally characterized enzyme from the bacterium, and this work broadens the knowledge of GH10 xylanases. Academic Press Inc. 2019-12 Article PeerReviewed Liew, Kok Jun and Ngooi, Chen Yi and Shamsir, Mohd. Shahir and Sani, Rajesh Kumar and Chong, Chun Shiong and Goh, Kian Mau (2019) Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA. Protein Expression and Purification, 164 . p. 105464. ISSN 1046-5928 http://dx.doi.org/10.1016/j.pep.2019.105464 Volume , December , Article number
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
topic QD Chemistry
spellingShingle QD Chemistry
Liew, Kok Jun
Ngooi, Chen Yi
Shamsir, Mohd. Shahir
Sani, Rajesh Kumar
Chong, Chun Shiong
Goh, Kian Mau
Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA
description Xylanases (EC 3.2.1.8) are essential enzymes due to their applications in various industries such as textile, animal feed, paper and pulp, and biofuel industries. Halo-thermophilic Rhodothermaceae bacterium RA was previously isolated from a hot spring in Malaysia. Genomic analysis revealed that this bacterium is likely to be a new genus of the family Rhodothermaceae. In this study, a xylanase gene (1140 bp) that encoded 379 amino acids from the bacterium was cloned and expressed in Escherichia coli BL21(DE3). Based on InterProScan, this enzyme XynRA1 contained a GH10 domain and a signal peptide sequence. XynRA1 shared low similarity with the currently known xylanases (the closest is 57.2–65.4% to Gemmatimonadetes spp.). The purified XynRA1 achieved maximum activity at pH 8 and 60 °C. The protein molecular weight was 43.1 kDa XynRA1 exhibited an activity half-life (t1/2) of 1 h at 60 °C and remained stable at 50 °C throughout the experiment. However, it was NaCl intolerant, and various types of salt reduced the activity. This enzyme effectively hydrolyzed xylan (beechwood, oat spelt, and Palmaria palmata) and xylodextrin (xylotriose, xylotetraose, xylopentaose, and xylohexaose) to produce predominantly xylobiose. This xylanase is the first functionally characterized enzyme from the bacterium, and this work broadens the knowledge of GH10 xylanases.
format Article
author Liew, Kok Jun
Ngooi, Chen Yi
Shamsir, Mohd. Shahir
Sani, Rajesh Kumar
Chong, Chun Shiong
Goh, Kian Mau
author_facet Liew, Kok Jun
Ngooi, Chen Yi
Shamsir, Mohd. Shahir
Sani, Rajesh Kumar
Chong, Chun Shiong
Goh, Kian Mau
author_sort Liew, Kok Jun
title Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA
title_short Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA
title_full Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA
title_fullStr Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA
title_full_unstemmed Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA
title_sort heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from rhodothermaceae bacterium ra
publisher Academic Press Inc.
publishDate 2019
url http://eprints.utm.my/id/eprint/89065/
http://dx.doi.org/10.1016/j.pep.2019.105464
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score 13.211869

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