Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa

Abstract Endoglucanase producing bacteria were isolated from Egyptian soils and the most active bacterial strain was identified as Bacillus subtilis strain Fatma/1. Plackett–Burman statistical design was carried out to assess the effect of seven process variables on endoglucanase production. Carboxy...

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Autores principales:	Atef M. Ibrahim, Ragaa A. Hamouda, Noura El-Ahmady El-Naggar, Fatma M. Al-Shakankery
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d3b1393514844b3998aa14f98036e880
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spelling	oai:doaj.org-article:d3b1393514844b3998aa14f98036e8802021-12-02T14:49:26ZBioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa10.1038/s41598-021-87901-92045-2322https://doaj.org/article/d3b1393514844b3998aa14f98036e8802021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-87901-9https://doaj.org/toc/2045-2322Abstract Endoglucanase producing bacteria were isolated from Egyptian soils and the most active bacterial strain was identified as Bacillus subtilis strain Fatma/1. Plackett–Burman statistical design was carried out to assess the effect of seven process variables on endoglucanase production. Carboxymethyl cellulose (CMC), yeast extract and peptone were the most significant variables that enhanced the endoglucanase production and thus were selected for further optimization using face-centered central composite design. The highest yield of endoglucanase (32.37 U/mL) was obtained in run no. 9, using 18 g/L CMC, 8 g/L peptone, 7 g/L yeast extract and 0.1 g/L FeSO4.7H2O. The optimized medium showed about eightfold increase in endoglucanase production compared to the unoptimized medium. The produced crude enzyme was further purified by ammonium sulfate precipitation, then DEAE-Sepharose CL6B column. The purified enzyme was shown to have a molecular weight of 37 kDa. The enzyme showed maximum activity at pH 8.0, temperature of 50 °C, incubation time of 60 min. The half-life time (T1/2) was 139.53 min at 50 °C, while being 82.67 min at 60 °C. Endoglucanase at concentration of 12 U/mL effectively removed 84.61% of biofilm matrix of Pseudomonas aeruginosa with marked reduction in carbohydrate content of the biofilm from 63.4 to 7.9 μg.Atef M. IbrahimRagaa A. HamoudaNoura El-Ahmady El-NaggarFatma M. Al-ShakankeryNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-24 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Atef M. Ibrahim Ragaa A. Hamouda Noura El-Ahmady El-Naggar Fatma M. Al-Shakankery Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa
description	Abstract Endoglucanase producing bacteria were isolated from Egyptian soils and the most active bacterial strain was identified as Bacillus subtilis strain Fatma/1. Plackett–Burman statistical design was carried out to assess the effect of seven process variables on endoglucanase production. Carboxymethyl cellulose (CMC), yeast extract and peptone were the most significant variables that enhanced the endoglucanase production and thus were selected for further optimization using face-centered central composite design. The highest yield of endoglucanase (32.37 U/mL) was obtained in run no. 9, using 18 g/L CMC, 8 g/L peptone, 7 g/L yeast extract and 0.1 g/L FeSO4.7H2O. The optimized medium showed about eightfold increase in endoglucanase production compared to the unoptimized medium. The produced crude enzyme was further purified by ammonium sulfate precipitation, then DEAE-Sepharose CL6B column. The purified enzyme was shown to have a molecular weight of 37 kDa. The enzyme showed maximum activity at pH 8.0, temperature of 50 °C, incubation time of 60 min. The half-life time (T1/2) was 139.53 min at 50 °C, while being 82.67 min at 60 °C. Endoglucanase at concentration of 12 U/mL effectively removed 84.61% of biofilm matrix of Pseudomonas aeruginosa with marked reduction in carbohydrate content of the biofilm from 63.4 to 7.9 μg.
format	article
author	Atef M. Ibrahim Ragaa A. Hamouda Noura El-Ahmady El-Naggar Fatma M. Al-Shakankery
author_facet	Atef M. Ibrahim Ragaa A. Hamouda Noura El-Ahmady El-Naggar Fatma M. Al-Shakankery
author_sort	Atef M. Ibrahim
title	Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa
title_short	Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa
title_full	Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa
title_fullStr	Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa
title_full_unstemmed	Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa
title_sort	bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of pseudomonas aeruginosa
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/d3b1393514844b3998aa14f98036e880
work_keys_str_mv	AT atefmibrahim bioprocessdevelopmentforenhancedendoglucanaseproductionbynewlyisolatedbacteriapurificationcharacterizationandinvitroefficacyasantibiofilmofpseudomonasaeruginosa AT ragaaahamouda bioprocessdevelopmentforenhancedendoglucanaseproductionbynewlyisolatedbacteriapurificationcharacterizationandinvitroefficacyasantibiofilmofpseudomonasaeruginosa AT nouraelahmadyelnaggar bioprocessdevelopmentforenhancedendoglucanaseproductionbynewlyisolatedbacteriapurificationcharacterizationandinvitroefficacyasantibiofilmofpseudomonasaeruginosa AT fatmamalshakankery bioprocessdevelopmentforenhancedendoglucanaseproductionbynewlyisolatedbacteriapurificationcharacterizationandinvitroefficacyasantibiofilmofpseudomonasaeruginosa
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Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa

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