Poly(L-lactide)-degrading enzyme from laceyella sacchari LP175: Cloning, sequencing, expression, characterization and its hydrolysis of poly(L-lactide) polymer

© 2019, All Right reserved. In this study, the poly-(L-lactide) (PLLA)-degrading enzyme from the thermophilic filamentous bacterium, Laceyella sacchari LP175 was characterized for application in biological recycling process. The gene encoding the PLLA-degrading gene (plla_lp175) was cloned and expre...

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Main Authors: Thanasak Lomthong, Marie Guicherd, Gianluca Cioci, Sophie Duquesne, Alain Marty, Saisamorn Lumyong, Vichien Kitpreechavanich
Format: Journal
Published: 2019
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http://cmuir.cmu.ac.th/jspui/handle/6653943832/66598
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-665982019-09-16T13:00:09Z Poly(L-lactide)-degrading enzyme from laceyella sacchari LP175: Cloning, sequencing, expression, characterization and its hydrolysis of poly(L-lactide) polymer Thanasak Lomthong Marie Guicherd Gianluca Cioci Sophie Duquesne Alain Marty Saisamorn Lumyong Vichien Kitpreechavanich Biochemistry, Genetics and Molecular Biology Chemistry Materials Science Mathematics Physics and Astronomy © 2019, All Right reserved. In this study, the poly-(L-lactide) (PLLA)-degrading enzyme from the thermophilic filamentous bacterium, Laceyella sacchari LP175 was characterized for application in biological recycling process. The gene encoding the PLLA-degrading gene (plla_lp175) was cloned and expressed in Escherichia coli. The native protein comprises 383 amino acids with a molecular mass and pI of 39.45 kDa and 8.26, respectively. The recombinant protein was purified by one-step purification, and was found to have a molecular weight of 28 kDa. Functional expression of PLLA_LP175 in E. coli with a pMAL-c5X vector enhanced the expression of PLLA-degrading enzyme up to 756 U/mg of protein, which is a 2.3-fold increase compared to the native strain. The purified recombinant protein is active in the range of pH 7.0-9.0 and 45-60 °C, with optimum activity at pH 9.0 and 60 °C. The recombinant enzyme could hydrolyse PLLA objects to a different extent depending on PLLA composition, at pH 9.0 and 50 °C within 24 h. Monomeric lactic acid was detected as the product from enzymatic degradation of PLLA objects. A scanning electron micrograph, showing a rough surface with holes of all PLLA objects after treatments with the recombinant enzyme, confirmed the ability of this enzyme to degrade PLLA objects. 2019-09-16T12:47:48Z 2019-09-16T12:47:48Z 2019-05-01 Journal 01252526 2-s2.0-85071146117 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85071146117&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/66598
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Biochemistry, Genetics and Molecular Biology
Chemistry
Materials Science
Mathematics
Physics and Astronomy
spellingShingle Biochemistry, Genetics and Molecular Biology
Chemistry
Materials Science
Mathematics
Physics and Astronomy
Thanasak Lomthong
Marie Guicherd
Gianluca Cioci
Sophie Duquesne
Alain Marty
Saisamorn Lumyong
Vichien Kitpreechavanich
Poly(L-lactide)-degrading enzyme from laceyella sacchari LP175: Cloning, sequencing, expression, characterization and its hydrolysis of poly(L-lactide) polymer
description © 2019, All Right reserved. In this study, the poly-(L-lactide) (PLLA)-degrading enzyme from the thermophilic filamentous bacterium, Laceyella sacchari LP175 was characterized for application in biological recycling process. The gene encoding the PLLA-degrading gene (plla_lp175) was cloned and expressed in Escherichia coli. The native protein comprises 383 amino acids with a molecular mass and pI of 39.45 kDa and 8.26, respectively. The recombinant protein was purified by one-step purification, and was found to have a molecular weight of 28 kDa. Functional expression of PLLA_LP175 in E. coli with a pMAL-c5X vector enhanced the expression of PLLA-degrading enzyme up to 756 U/mg of protein, which is a 2.3-fold increase compared to the native strain. The purified recombinant protein is active in the range of pH 7.0-9.0 and 45-60 °C, with optimum activity at pH 9.0 and 60 °C. The recombinant enzyme could hydrolyse PLLA objects to a different extent depending on PLLA composition, at pH 9.0 and 50 °C within 24 h. Monomeric lactic acid was detected as the product from enzymatic degradation of PLLA objects. A scanning electron micrograph, showing a rough surface with holes of all PLLA objects after treatments with the recombinant enzyme, confirmed the ability of this enzyme to degrade PLLA objects.
format Journal
author Thanasak Lomthong
Marie Guicherd
Gianluca Cioci
Sophie Duquesne
Alain Marty
Saisamorn Lumyong
Vichien Kitpreechavanich
author_facet Thanasak Lomthong
Marie Guicherd
Gianluca Cioci
Sophie Duquesne
Alain Marty
Saisamorn Lumyong
Vichien Kitpreechavanich
author_sort Thanasak Lomthong
title Poly(L-lactide)-degrading enzyme from laceyella sacchari LP175: Cloning, sequencing, expression, characterization and its hydrolysis of poly(L-lactide) polymer
title_short Poly(L-lactide)-degrading enzyme from laceyella sacchari LP175: Cloning, sequencing, expression, characterization and its hydrolysis of poly(L-lactide) polymer
title_full Poly(L-lactide)-degrading enzyme from laceyella sacchari LP175: Cloning, sequencing, expression, characterization and its hydrolysis of poly(L-lactide) polymer
title_fullStr Poly(L-lactide)-degrading enzyme from laceyella sacchari LP175: Cloning, sequencing, expression, characterization and its hydrolysis of poly(L-lactide) polymer
title_full_unstemmed Poly(L-lactide)-degrading enzyme from laceyella sacchari LP175: Cloning, sequencing, expression, characterization and its hydrolysis of poly(L-lactide) polymer
title_sort poly(l-lactide)-degrading enzyme from laceyella sacchari lp175: cloning, sequencing, expression, characterization and its hydrolysis of poly(l-lactide) polymer
publishDate 2019
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85071146117&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/66598
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