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Article

Enhanced Thermostability and Enzymatic Activity of cel6A Variants from Thermobifida fusca by Empirical Domain Engineering

1
Department of Biotechnology, Mirpur University of Science and Technology (MUST), Mirpur (AJK) 10250, Pakistan
2
Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore 54590, Pakistan
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Department of Human Genetics and Molecular Biology, University of Health Sciences, Lahore 54590, Pakistan
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Department of Pharmaceutical and Pharmacological Sciences, Rega Institute for Medical Research, Medicinal Chemistry, University of Leuven, B-3000 Leuven, Belgium
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School of Biological Sciences, University of the Punjab, Lahore 54590, Pakistan
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Department of Biotechnology, Forman Christian College University, Lahore 54590, Pakistan
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Shaheed Zulfiqar Ali Bhutto Medical University, Islamabad 44000, Pakistan
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Department of Biology, College of Sciences, King Khalid University, Abha 61413, Saudi Arabia
*
Author to whom correspondence should be addressed.
Biology 2020, 9(8), 214; https://doi.org/10.3390/biology9080214
Received: 26 June 2020 / Revised: 29 July 2020 / Accepted: 2 August 2020 / Published: 7 August 2020
(This article belongs to the Section Biotechnology)
Cellulases are a set of lignocellulolytic enzymes, capable of producing eco-friendly low-cost renewable bioethanol. However, low stability and hydrolytic activity limit their wide-scale applicability at the industrial scale. In this work, we report the domain engineering of endoglucanase (cel6A) of Thermobifida fusca to improve their catalytic activity and thermal stability. Later, enzymatic activity and thermostability of the most efficient variant named as cel6A.CBC was analyzed by molecular dynamics simulations. This variant demonstrated profound activity against soluble and insoluble cellulosic substrates like filter paper, alkali-treated bagasse, regenerated amorphous cellulose (RAC), and bacterial microcrystalline cellulose. The variant cel6A.CBC showed the highest catalysis of carboxymethyl cellulose (CMC) and other related insoluble substrates at a pH of 6.0 and a temperature of 60 °C. Furthermore, a sound rationale was observed between experimental findings and molecular modeling of cel6A.CBC which revealed thermostability of cel6A.CBC at 26.85, 60.85, and 74.85 °C as well as structural flexibility at 126.85 °C. Therefore, a thermostable derivative of cel6A engineered in the present work has enhanced biological performance and can be a useful construct for the mass production of bioethanol from plant biomass. View Full-Text
Keywords: endoglucanase (cel6A); domain engineering; Thermobifida fusca; molecular dynamics simulations; thermostable enzymes endoglucanase (cel6A); domain engineering; Thermobifida fusca; molecular dynamics simulations; thermostable enzymes
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MDPI and ACS Style

Ali, I.; Rehman, H.M.; Mirza, M.U.; Akhtar, M.W.; Asghar, R.; Tariq, M.; Ahmed, R.; Tanveer, F.; Khalid, H.; Alghamdi, H.A.; Froeyen, M. Enhanced Thermostability and Enzymatic Activity of cel6A Variants from Thermobifida fusca by Empirical Domain Engineering. Biology 2020, 9, 214. https://doi.org/10.3390/biology9080214

AMA Style

Ali I, Rehman HM, Mirza MU, Akhtar MW, Asghar R, Tariq M, Ahmed R, Tanveer F, Khalid H, Alghamdi HA, Froeyen M. Enhanced Thermostability and Enzymatic Activity of cel6A Variants from Thermobifida fusca by Empirical Domain Engineering. Biology. 2020; 9(8):214. https://doi.org/10.3390/biology9080214

Chicago/Turabian Style

Ali, Imran, Hafiz M. Rehman, Muhammad U. Mirza, Muhammad W. Akhtar, Rehana Asghar, Muhammad Tariq, Rashid Ahmed, Fatima Tanveer, Hina Khalid, Huda A. Alghamdi, and Matheus Froeyen. 2020. "Enhanced Thermostability and Enzymatic Activity of cel6A Variants from Thermobifida fusca by Empirical Domain Engineering" Biology 9, no. 8: 214. https://doi.org/10.3390/biology9080214

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