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Ancestral sequence reconstruction of ancient lipase from family I.3 bacterial lipolytic enzymes

Family I.3 lipase is distinguished from other families by the amino acid sequence and secretion mechanism. Little is known about the evolutionary process driving these differences. This study attempt to understand how the diverse temperature stabilities of bacterial lipases from family I.3 evolved....

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Main Authors: Mohd Rozi, Mohamad Farihan Afnan, Raja Abd. Rahman, Raja Noor Zaliha, Thean, Adam Chor Leow, Mohamad Ali, Mohd Shukuri
Format: Article
Published: Elsevier 2022
Online Access:http://psasir.upm.edu.my/id/eprint/100230/
https://www.sciencedirect.com/science/article/pii/S1055790321003146
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spelling my.upm.eprints.1002302024-07-11T04:06:35Z http://psasir.upm.edu.my/id/eprint/100230/ Ancestral sequence reconstruction of ancient lipase from family I.3 bacterial lipolytic enzymes Mohd Rozi, Mohamad Farihan Afnan Raja Abd. Rahman, Raja Noor Zaliha Thean, Adam Chor Leow Mohamad Ali, Mohd Shukuri Family I.3 lipase is distinguished from other families by the amino acid sequence and secretion mechanism. Little is known about the evolutionary process driving these differences. This study attempt to understand how the diverse temperature stabilities of bacterial lipases from family I.3 evolved. To achieve that, eighty-three protein sequences sharing a minimum 30% sequence identity with Antarctic Pseudomonas sp. AMS8 lipase were used to infer phylogenetic tree. Using ancestral sequence reconstruction (ASR) technique, the last universal common ancestor (LUCA) sequence of family I.3 was reconstructed. A gene encoding LUCA was synthesized, cloned and expressed as inclusion bodies in E. coli system. Insoluble form of LUCA was refolded using urea dilution method and then purified using affinity chromatography. The purified LUCA exhibited an optimum temperature and pH at 70 ℃ and 10 respectively. Various metal ions increased or retained the activity of LUCA. LUCA also demonstrated tolerance towards various organic solvents in 25% v/v concentration. The finding from this study could support the understanding on temperature and environment during ancient time. In overall, reconstructed ancestral enzymes have improved physicochemical properties that make them suitable for industrial applications and ASR technique can be employed as a general technique for enzyme engineering. Elsevier 2022-03 Article PeerReviewed Mohd Rozi, Mohamad Farihan Afnan and Raja Abd. Rahman, Raja Noor Zaliha and Thean, Adam Chor Leow and Mohamad Ali, Mohd Shukuri (2022) Ancestral sequence reconstruction of ancient lipase from family I.3 bacterial lipolytic enzymes. Molecular Phylogenetics and Evolution, 168. art. no. 107381. pp. 1-10. ISSN 1055-7903; ESSN: 1095-9513 https://www.sciencedirect.com/science/article/pii/S1055790321003146 10.1016/j.ympev.2021.107381
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 Family I.3 lipase is distinguished from other families by the amino acid sequence and secretion mechanism. Little is known about the evolutionary process driving these differences. This study attempt to understand how the diverse temperature stabilities of bacterial lipases from family I.3 evolved. To achieve that, eighty-three protein sequences sharing a minimum 30% sequence identity with Antarctic Pseudomonas sp. AMS8 lipase were used to infer phylogenetic tree. Using ancestral sequence reconstruction (ASR) technique, the last universal common ancestor (LUCA) sequence of family I.3 was reconstructed. A gene encoding LUCA was synthesized, cloned and expressed as inclusion bodies in E. coli system. Insoluble form of LUCA was refolded using urea dilution method and then purified using affinity chromatography. The purified LUCA exhibited an optimum temperature and pH at 70 ℃ and 10 respectively. Various metal ions increased or retained the activity of LUCA. LUCA also demonstrated tolerance towards various organic solvents in 25% v/v concentration. The finding from this study could support the understanding on temperature and environment during ancient time. In overall, reconstructed ancestral enzymes have improved physicochemical properties that make them suitable for industrial applications and ASR technique can be employed as a general technique for enzyme engineering.
format Article
author Mohd Rozi, Mohamad Farihan Afnan
Raja Abd. Rahman, Raja Noor Zaliha
Thean, Adam Chor Leow
Mohamad Ali, Mohd Shukuri
spellingShingle Mohd Rozi, Mohamad Farihan Afnan
Raja Abd. Rahman, Raja Noor Zaliha
Thean, Adam Chor Leow
Mohamad Ali, Mohd Shukuri
Ancestral sequence reconstruction of ancient lipase from family I.3 bacterial lipolytic enzymes
author_facet Mohd Rozi, Mohamad Farihan Afnan
Raja Abd. Rahman, Raja Noor Zaliha
Thean, Adam Chor Leow
Mohamad Ali, Mohd Shukuri
author_sort Mohd Rozi, Mohamad Farihan Afnan
title Ancestral sequence reconstruction of ancient lipase from family I.3 bacterial lipolytic enzymes
title_short Ancestral sequence reconstruction of ancient lipase from family I.3 bacterial lipolytic enzymes
title_full Ancestral sequence reconstruction of ancient lipase from family I.3 bacterial lipolytic enzymes
title_fullStr Ancestral sequence reconstruction of ancient lipase from family I.3 bacterial lipolytic enzymes
title_full_unstemmed Ancestral sequence reconstruction of ancient lipase from family I.3 bacterial lipolytic enzymes
title_sort ancestral sequence reconstruction of ancient lipase from family i.3 bacterial lipolytic enzymes
publisher Elsevier
publishDate 2022
url http://psasir.upm.edu.my/id/eprint/100230/
https://www.sciencedirect.com/science/article/pii/S1055790321003146
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score 13.211869

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