Polycistronic Expression System for Pichia pastoris Composed of Chitino- and Chitosanolytic Enzymes
Chitin is one of the most abundant biopolymers. Due to its recalcitrant nature and insolubility in accessible solvents, it is often considered waste and not a bioresource. The products of chitin modification such as chitosan and chitooligosaccharides are highly sought, but their preparation is a cha...
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doaj-3f79277ace7942af9ca96495d07b45d32021-08-18T09:48:38ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852021-08-01910.3389/fbioe.2021.710922710922Polycistronic Expression System for Pichia pastoris Composed of Chitino- and Chitosanolytic EnzymesMichal B. Kaczmarek0Michal B. Kaczmarek1Katarzyna Struszczyk-Swita2Meng Xiao3Mirosława Szczęsna-Antczak4Tadeusz Antczak5Magdalena Gierszewska6Alexander Steinbüchel7Maurycy Daroch8School of Environment and Energy, Peking University Shenzhen Graduate School, Shenzhen, ChinaInstitute of Molecular and Industrial Biotechnology, Lodz University of Technology, Lodz, PolandInstitute of Molecular and Industrial Biotechnology, Lodz University of Technology, Lodz, PolandSchool of Environment and Energy, Peking University Shenzhen Graduate School, Shenzhen, ChinaInstitute of Molecular and Industrial Biotechnology, Lodz University of Technology, Lodz, PolandInstitute of Molecular and Industrial Biotechnology, Lodz University of Technology, Lodz, PolandDepartment of Physical Chemistry and Physicochemistry of Polymers, Faculty of Chemistry, Nicolaus Copernicus University in Toruń, Toruń, PolandInternational Center for Research on Innovative Biobased Materials (ICRI-BioM), International Research Agenda, Lodz University of Technology, Lodz, PolandSchool of Environment and Energy, Peking University Shenzhen Graduate School, Shenzhen, ChinaChitin is one of the most abundant biopolymers. Due to its recalcitrant nature and insolubility in accessible solvents, it is often considered waste and not a bioresource. The products of chitin modification such as chitosan and chitooligosaccharides are highly sought, but their preparation is a challenging process, typically performed with thermochemical methods that lack specificities and generate hazardous waste. Enzymatic treatment is a promising alternative to these methods, but the preparation of multiple biocatalysts is costly. In this manuscript, we biochemically characterised chitin deacetylases of Mucor circinelloides IBT-83 and utilised one of them for the construction of the first eukaryotic, polycistronic expression system employing self-processing 2A sequences. The three chitin-processing enzymes; chitin deacetylase of M. circinelloides IBT-83, chitinase from Thermomyces lanuginosus, and chitosanase from Aspergillus fumigatus were expressed under the control of the same promoter in methylotrophic yeast Pichia pastoris and characterised for their synergistic action towards their respective substrates.https://www.frontiersin.org/articles/10.3389/fbioe.2021.710922/fullchitinchitosanenzymatic modificationpolycistronic expressionself-processing 2A sequencechitin deacetylase |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Michal B. Kaczmarek Michal B. Kaczmarek Katarzyna Struszczyk-Swita Meng Xiao Mirosława Szczęsna-Antczak Tadeusz Antczak Magdalena Gierszewska Alexander Steinbüchel Maurycy Daroch |
spellingShingle |
Michal B. Kaczmarek Michal B. Kaczmarek Katarzyna Struszczyk-Swita Meng Xiao Mirosława Szczęsna-Antczak Tadeusz Antczak Magdalena Gierszewska Alexander Steinbüchel Maurycy Daroch Polycistronic Expression System for Pichia pastoris Composed of Chitino- and Chitosanolytic Enzymes Frontiers in Bioengineering and Biotechnology chitin chitosan enzymatic modification polycistronic expression self-processing 2A sequence chitin deacetylase |
author_facet |
Michal B. Kaczmarek Michal B. Kaczmarek Katarzyna Struszczyk-Swita Meng Xiao Mirosława Szczęsna-Antczak Tadeusz Antczak Magdalena Gierszewska Alexander Steinbüchel Maurycy Daroch |
author_sort |
Michal B. Kaczmarek |
title |
Polycistronic Expression System for Pichia pastoris Composed of Chitino- and Chitosanolytic Enzymes |
title_short |
Polycistronic Expression System for Pichia pastoris Composed of Chitino- and Chitosanolytic Enzymes |
title_full |
Polycistronic Expression System for Pichia pastoris Composed of Chitino- and Chitosanolytic Enzymes |
title_fullStr |
Polycistronic Expression System for Pichia pastoris Composed of Chitino- and Chitosanolytic Enzymes |
title_full_unstemmed |
Polycistronic Expression System for Pichia pastoris Composed of Chitino- and Chitosanolytic Enzymes |
title_sort |
polycistronic expression system for pichia pastoris composed of chitino- and chitosanolytic enzymes |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Bioengineering and Biotechnology |
issn |
2296-4185 |
publishDate |
2021-08-01 |
description |
Chitin is one of the most abundant biopolymers. Due to its recalcitrant nature and insolubility in accessible solvents, it is often considered waste and not a bioresource. The products of chitin modification such as chitosan and chitooligosaccharides are highly sought, but their preparation is a challenging process, typically performed with thermochemical methods that lack specificities and generate hazardous waste. Enzymatic treatment is a promising alternative to these methods, but the preparation of multiple biocatalysts is costly. In this manuscript, we biochemically characterised chitin deacetylases of Mucor circinelloides IBT-83 and utilised one of them for the construction of the first eukaryotic, polycistronic expression system employing self-processing 2A sequences. The three chitin-processing enzymes; chitin deacetylase of M. circinelloides IBT-83, chitinase from Thermomyces lanuginosus, and chitosanase from Aspergillus fumigatus were expressed under the control of the same promoter in methylotrophic yeast Pichia pastoris and characterised for their synergistic action towards their respective substrates. |
topic |
chitin chitosan enzymatic modification polycistronic expression self-processing 2A sequence chitin deacetylase |
url |
https://www.frontiersin.org/articles/10.3389/fbioe.2021.710922/full |
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