Recombinant cellobiohydrolase of Myceliophthora thermophila: characterization and applicability in cellulose saccharification

Abstract A codon optimized cellobiohydrolase (CBH) encoding synthetic gene of 1188 bp from a thermophilic mold Myceliophthora thermophila (MtCel6A) was cloned and heterologously expressed in Escherichia coli for the first time. In silico analysis suggested that MtCel6A is a GH6 CBH and belongs to CB...

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Autores principales:	Anica Dadwal, Shilpa Sharma, Tulasi Satyanarayana
Formato:	article
Lenguaje:	EN
Publicado:	SpringerOpen 2021
Materias:	Thermostable cellulose Recombinant cellobiohydrolase Alkalistable Solvent tolerance Extremozyme Cellulose saccharification Biotechnology TP248.13-248.65 Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/b765d5e563634a91be867b6147a28a3d
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spelling	oai:doaj.org-article:b765d5e563634a91be867b6147a28a3d2021-11-07T12:22:00ZRecombinant cellobiohydrolase of Myceliophthora thermophila: characterization and applicability in cellulose saccharification10.1186/s13568-021-01311-82191-0855https://doaj.org/article/b765d5e563634a91be867b6147a28a3d2021-11-01T00:00:00Zhttps://doi.org/10.1186/s13568-021-01311-8https://doaj.org/toc/2191-0855Abstract A codon optimized cellobiohydrolase (CBH) encoding synthetic gene of 1188 bp from a thermophilic mold Myceliophthora thermophila (MtCel6A) was cloned and heterologously expressed in Escherichia coli for the first time. In silico analysis suggested that MtCel6A is a GH6 CBH and belongs to CBHII family, which is structurally similar to Cel6A of Humicola insolens. The recombinant MtCel6A is expressed as active inclusion bodies, and the molecular mass of the purified enzyme is ~ 45 kDa. The rMtCel6A is active in a wide range of pH (4–12) and temperatures (40–100 °C) with optima at pH 10.0 and 60 °C. It exhibits T1/2 of 6.0 and 1.0 h at 60 and 90 °C, respectively. The rMtCel6A is an extremozyme with organic solvent, salt and alkali tolerance. The Km, Vmax, kcat and kcat/Km values of the enzyme are 3.2 mg mL−1, 222.2 μmol mg−1 min−1, 2492 s−1 and 778.7 s−1 mg−1 mL−1, respectively. The product analysis of rMtCel6A confirmed that it is an exoenzyme that acts from the non-reducing end of cellulose. The addition of rMtCel6A to the commercial cellulase mix (Cellic CTec2) led to 1.9-fold increase in saccharification of the pre-treated sugarcane bagasse. The rMtCel6A is a potential CBH that finds utility in industrial processes such as in bioethanol, paper pulp and textile industries. Graphical AbstractAnica DadwalShilpa SharmaTulasi SatyanarayanaSpringerOpenarticleThermostable celluloseRecombinant cellobiohydrolaseAlkalistableSolvent toleranceExtremozymeCellulose saccharificationBiotechnologyTP248.13-248.65MicrobiologyQR1-502ENAMB Express, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Thermostable cellulose Recombinant cellobiohydrolase Alkalistable Solvent tolerance Extremozyme Cellulose saccharification Biotechnology TP248.13-248.65 Microbiology QR1-502
spellingShingle	Thermostable cellulose Recombinant cellobiohydrolase Alkalistable Solvent tolerance Extremozyme Cellulose saccharification Biotechnology TP248.13-248.65 Microbiology QR1-502 Anica Dadwal Shilpa Sharma Tulasi Satyanarayana Recombinant cellobiohydrolase of Myceliophthora thermophila: characterization and applicability in cellulose saccharification
description	Abstract A codon optimized cellobiohydrolase (CBH) encoding synthetic gene of 1188 bp from a thermophilic mold Myceliophthora thermophila (MtCel6A) was cloned and heterologously expressed in Escherichia coli for the first time. In silico analysis suggested that MtCel6A is a GH6 CBH and belongs to CBHII family, which is structurally similar to Cel6A of Humicola insolens. The recombinant MtCel6A is expressed as active inclusion bodies, and the molecular mass of the purified enzyme is ~ 45 kDa. The rMtCel6A is active in a wide range of pH (4–12) and temperatures (40–100 °C) with optima at pH 10.0 and 60 °C. It exhibits T1/2 of 6.0 and 1.0 h at 60 and 90 °C, respectively. The rMtCel6A is an extremozyme with organic solvent, salt and alkali tolerance. The Km, Vmax, kcat and kcat/Km values of the enzyme are 3.2 mg mL−1, 222.2 μmol mg−1 min−1, 2492 s−1 and 778.7 s−1 mg−1 mL−1, respectively. The product analysis of rMtCel6A confirmed that it is an exoenzyme that acts from the non-reducing end of cellulose. The addition of rMtCel6A to the commercial cellulase mix (Cellic CTec2) led to 1.9-fold increase in saccharification of the pre-treated sugarcane bagasse. The rMtCel6A is a potential CBH that finds utility in industrial processes such as in bioethanol, paper pulp and textile industries. Graphical Abstract
format	article
author	Anica Dadwal Shilpa Sharma Tulasi Satyanarayana
author_facet	Anica Dadwal Shilpa Sharma Tulasi Satyanarayana
author_sort	Anica Dadwal
title	Recombinant cellobiohydrolase of Myceliophthora thermophila: characterization and applicability in cellulose saccharification
title_short	Recombinant cellobiohydrolase of Myceliophthora thermophila: characterization and applicability in cellulose saccharification
title_full	Recombinant cellobiohydrolase of Myceliophthora thermophila: characterization and applicability in cellulose saccharification
title_fullStr	Recombinant cellobiohydrolase of Myceliophthora thermophila: characterization and applicability in cellulose saccharification
title_full_unstemmed	Recombinant cellobiohydrolase of Myceliophthora thermophila: characterization and applicability in cellulose saccharification
title_sort	recombinant cellobiohydrolase of myceliophthora thermophila: characterization and applicability in cellulose saccharification
publisher	SpringerOpen
publishDate	2021
url	https://doaj.org/article/b765d5e563634a91be867b6147a28a3d
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Recombinant cellobiohydrolase of Myceliophthora thermophila: characterization and applicability in cellulose saccharification

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