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Combined Optimization of Codon Usage and Glycine Supplementation Enhances the Extracellular Production of a β-Cyclodextrin Glycosyltransferase from Bacillus sp. NR5 UPM in Escherichia coli

Two optimization strategies, codon usage modification and glycine supplementation, were adopted to improve the extracellular production of Bacillus sp. NR5 UPM β-cyclodextrin glycosyltransferase (CGT-BS) in recombinant Escherichia coli. Several rare codons were eliminated and replaced with the ones...

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Autores principales: Nik-Pa, Nik Ida Mardiana, Sobri, Mohamad Farhan Mohamad, Abd-Aziz, Suraini, Ibrahim, Mohamad Faizal, Kamal Bahrin, Ezyana, Mohammed Alitheen, Noorjahan Banu, Ramli, Norhayati
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7313058/
https://www.ncbi.nlm.nih.gov/pubmed/32486212
http://dx.doi.org/10.3390/ijms21113919
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author Nik-Pa, Nik Ida Mardiana
Sobri, Mohamad Farhan Mohamad
Abd-Aziz, Suraini
Ibrahim, Mohamad Faizal
Kamal Bahrin, Ezyana
Mohammed Alitheen, Noorjahan Banu
Ramli, Norhayati
author_facet Nik-Pa, Nik Ida Mardiana
Sobri, Mohamad Farhan Mohamad
Abd-Aziz, Suraini
Ibrahim, Mohamad Faizal
Kamal Bahrin, Ezyana
Mohammed Alitheen, Noorjahan Banu
Ramli, Norhayati
author_sort Nik-Pa, Nik Ida Mardiana
collection PubMed
description Two optimization strategies, codon usage modification and glycine supplementation, were adopted to improve the extracellular production of Bacillus sp. NR5 UPM β-cyclodextrin glycosyltransferase (CGT-BS) in recombinant Escherichia coli. Several rare codons were eliminated and replaced with the ones favored by E. coli cells, resulting in an increased codon adaptation index (CAI) from 0.67 to 0.78. The cultivation of the codon modified recombinant E. coli following optimization of glycine supplementation enhanced the secretion of β-CGTase activity up to 2.2-fold at 12 h of cultivation as compared to the control. β-CGTase secreted into the culture medium by the transformant reached 65.524 U/mL at post-induction temperature of 37 °C with addition of 1.2 mM glycine and induced at 2 h of cultivation. A 20.1-fold purity of the recombinant β-CGTase was obtained when purified through a combination of diafiltration and nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography. This combined strategy doubled the extracellular β-CGTase production when compared to the single approach, hence offering the potential of enhancing the expression of extracellular enzymes, particularly β-CGTase by the recombinant E. coli.
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spelling pubmed-73130582020-06-29 Combined Optimization of Codon Usage and Glycine Supplementation Enhances the Extracellular Production of a β-Cyclodextrin Glycosyltransferase from Bacillus sp. NR5 UPM in Escherichia coli Nik-Pa, Nik Ida Mardiana Sobri, Mohamad Farhan Mohamad Abd-Aziz, Suraini Ibrahim, Mohamad Faizal Kamal Bahrin, Ezyana Mohammed Alitheen, Noorjahan Banu Ramli, Norhayati Int J Mol Sci Article Two optimization strategies, codon usage modification and glycine supplementation, were adopted to improve the extracellular production of Bacillus sp. NR5 UPM β-cyclodextrin glycosyltransferase (CGT-BS) in recombinant Escherichia coli. Several rare codons were eliminated and replaced with the ones favored by E. coli cells, resulting in an increased codon adaptation index (CAI) from 0.67 to 0.78. The cultivation of the codon modified recombinant E. coli following optimization of glycine supplementation enhanced the secretion of β-CGTase activity up to 2.2-fold at 12 h of cultivation as compared to the control. β-CGTase secreted into the culture medium by the transformant reached 65.524 U/mL at post-induction temperature of 37 °C with addition of 1.2 mM glycine and induced at 2 h of cultivation. A 20.1-fold purity of the recombinant β-CGTase was obtained when purified through a combination of diafiltration and nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography. This combined strategy doubled the extracellular β-CGTase production when compared to the single approach, hence offering the potential of enhancing the expression of extracellular enzymes, particularly β-CGTase by the recombinant E. coli. MDPI 2020-05-30 /pmc/articles/PMC7313058/ /pubmed/32486212 http://dx.doi.org/10.3390/ijms21113919 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nik-Pa, Nik Ida Mardiana
Sobri, Mohamad Farhan Mohamad
Abd-Aziz, Suraini
Ibrahim, Mohamad Faizal
Kamal Bahrin, Ezyana
Mohammed Alitheen, Noorjahan Banu
Ramli, Norhayati
Combined Optimization of Codon Usage and Glycine Supplementation Enhances the Extracellular Production of a β-Cyclodextrin Glycosyltransferase from Bacillus sp. NR5 UPM in Escherichia coli
title Combined Optimization of Codon Usage and Glycine Supplementation Enhances the Extracellular Production of a β-Cyclodextrin Glycosyltransferase from Bacillus sp. NR5 UPM in Escherichia coli
title_full Combined Optimization of Codon Usage and Glycine Supplementation Enhances the Extracellular Production of a β-Cyclodextrin Glycosyltransferase from Bacillus sp. NR5 UPM in Escherichia coli
title_fullStr Combined Optimization of Codon Usage and Glycine Supplementation Enhances the Extracellular Production of a β-Cyclodextrin Glycosyltransferase from Bacillus sp. NR5 UPM in Escherichia coli
title_full_unstemmed Combined Optimization of Codon Usage and Glycine Supplementation Enhances the Extracellular Production of a β-Cyclodextrin Glycosyltransferase from Bacillus sp. NR5 UPM in Escherichia coli
title_short Combined Optimization of Codon Usage and Glycine Supplementation Enhances the Extracellular Production of a β-Cyclodextrin Glycosyltransferase from Bacillus sp. NR5 UPM in Escherichia coli
title_sort combined optimization of codon usage and glycine supplementation enhances the extracellular production of a β-cyclodextrin glycosyltransferase from bacillus sp. nr5 upm in escherichia coli
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7313058/
https://www.ncbi.nlm.nih.gov/pubmed/32486212
http://dx.doi.org/10.3390/ijms21113919
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