Staff View: A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9

A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9

Carboxylesterases are attractive biocatalysts for various industrial applications, especially hyperthermophilic carboxylesterases, due to their high tolerance toward extreme environments. Such ability confers many advantages, including cost-effectiveness and an increased manufacturing rate. In the c...

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Main Authors:	Mohd Johan, Ummie Umaiera, Raja Abd. Rahman, Raja Noor Zaliha, Ahmad Kamarudin, Nor Hafizah, Latip, Wahhida, Mohamad Ali, Mohd Shukuri
Format:	Article
Published:	Elsevier 2022
Online Access:	http://psasir.upm.edu.my/id/eprint/100244/ https://www.sciencedirect.com/science/article/pii/S0141813022022668
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spelling	my.upm.eprints.1002442024-03-18T05:07:27Z http://psasir.upm.edu.my/id/eprint/100244/ A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9 Mohd Johan, Ummie Umaiera Raja Abd. Rahman, Raja Noor Zaliha Ahmad Kamarudin, Nor Hafizah Latip, Wahhida Mohamad Ali, Mohd Shukuri Carboxylesterases are attractive biocatalysts for various industrial applications, especially hyperthermophilic carboxylesterases, due to their high tolerance toward extreme environments. Such ability confers many advantages, including cost-effectiveness and an increased manufacturing rate. In the current work, we first described the characterization of EstD9, a new carboxylesterase from thermophilic Anoxybacillus geothermalis D9. Sequence analysis of EstD9 revealed a significant identity (80 %) with thermophilic Est30 and a catalytic triad, composed of Ser93-His22-Asp193. As the protein sequence contained a conserved pentapeptide (GLSLG), EstD9 could be proposed as a new member of family XIII. The putative carboxylesterase was recombinantly expressed in E. coli BL21 (DE3) with a molecular mass of 28 kDa and successfully purified via affinity chromatography with recovery of 88.36 %. Using p-nitrophenyl butyrate, EstD9 presented excellent stability at high temperature range (70 °C–100 °C) and a broad pH tolerance (pH 6–9), with optimal activity at 80 °C and pH 7. Notably, EstD9 activity was stimulated in the presence of 1-propanol and DMSO with 107.8 % and 108.9 % relative activities, respectively. The purified EstD9 maintained 60 % residual activity after 30 min exposure to various surfactants and metal ions. Additionally, the inhibition studies demonstrated strong deactivation by phenylmethylsulfonyl fluoride, dithiothreitol, and β-mercaptoethanol. The estimated Tm value was 72.12 °C. Unlike typical carboxylesterases, in silico 3D model of EstD9 disclosed a topological α/β hydrolase fold with a small α-helix cap. The enzymatic properties of EstD9 suggest this enzyme to be a highly suitable catalyst for industrial bioprocesses under harsh conditions. Elsevier 2022-12-01 Article PeerReviewed Mohd Johan, Ummie Umaiera and Raja Abd. Rahman, Raja Noor Zaliha and Ahmad Kamarudin, Nor Hafizah and Latip, Wahhida and Mohamad Ali, Mohd Shukuri (2022) A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9. International Journal of Biological Macromolecules, 222 (pt. B). pp. 2486-2497. ISSN 0141-8130 https://www.sciencedirect.com/science/article/pii/S0141813022022668 10.1016/j.ijbiomac.2022.10.033
institution	Universiti Putra Malaysia
building	UPM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Putra Malaysia
content_source	UPM Institutional Repository
url_provider	http://psasir.upm.edu.my/
description	Carboxylesterases are attractive biocatalysts for various industrial applications, especially hyperthermophilic carboxylesterases, due to their high tolerance toward extreme environments. Such ability confers many advantages, including cost-effectiveness and an increased manufacturing rate. In the current work, we first described the characterization of EstD9, a new carboxylesterase from thermophilic Anoxybacillus geothermalis D9. Sequence analysis of EstD9 revealed a significant identity (80 %) with thermophilic Est30 and a catalytic triad, composed of Ser93-His22-Asp193. As the protein sequence contained a conserved pentapeptide (GLSLG), EstD9 could be proposed as a new member of family XIII. The putative carboxylesterase was recombinantly expressed in E. coli BL21 (DE3) with a molecular mass of 28 kDa and successfully purified via affinity chromatography with recovery of 88.36 %. Using p-nitrophenyl butyrate, EstD9 presented excellent stability at high temperature range (70 °C–100 °C) and a broad pH tolerance (pH 6–9), with optimal activity at 80 °C and pH 7. Notably, EstD9 activity was stimulated in the presence of 1-propanol and DMSO with 107.8 % and 108.9 % relative activities, respectively. The purified EstD9 maintained 60 % residual activity after 30 min exposure to various surfactants and metal ions. Additionally, the inhibition studies demonstrated strong deactivation by phenylmethylsulfonyl fluoride, dithiothreitol, and β-mercaptoethanol. The estimated Tm value was 72.12 °C. Unlike typical carboxylesterases, in silico 3D model of EstD9 disclosed a topological α/β hydrolase fold with a small α-helix cap. The enzymatic properties of EstD9 suggest this enzyme to be a highly suitable catalyst for industrial bioprocesses under harsh conditions.
format	Article
author	Mohd Johan, Ummie Umaiera Raja Abd. Rahman, Raja Noor Zaliha Ahmad Kamarudin, Nor Hafizah Latip, Wahhida Mohamad Ali, Mohd Shukuri
spellingShingle	Mohd Johan, Ummie Umaiera Raja Abd. Rahman, Raja Noor Zaliha Ahmad Kamarudin, Nor Hafizah Latip, Wahhida Mohamad Ali, Mohd Shukuri A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9
author_facet	Mohd Johan, Ummie Umaiera Raja Abd. Rahman, Raja Noor Zaliha Ahmad Kamarudin, Nor Hafizah Latip, Wahhida Mohamad Ali, Mohd Shukuri
author_sort	Mohd Johan, Ummie Umaiera
title	A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9
title_short	A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9
title_full	A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9
title_fullStr	A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9
title_full_unstemmed	A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9
title_sort	new hyper-thermostable carboxylesterase from anoxybacillus geothermalis d9
publisher	Elsevier
publishDate	2022
url	http://psasir.upm.edu.my/id/eprint/100244/ https://www.sciencedirect.com/science/article/pii/S0141813022022668
_version_	1794564289665695744
score	13.211869

A new hyper-thermostable carboxylesterase from Anoxybacillus geothermalis D9

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