Biochemical and Molecular Characterization of Five Bacillus Isolates Displaying Remarkable Carboxymethyl Cellulase Activities

Elhameed, Esraa Abd; Sayed, Alaa R. M.; Radwan, Tharwat; Hassan, Gamal M.

doi:10.1007/s00284-020-02135-8

Cited by 9 publications

(6 citation statements)

References 33 publications

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“…Vijayaraghavan et al (2016) also reported the temperature of 60 °C as optimum for cellulase activity by a bacterium, Bacillus halodurans IND18 isolated from cow dung. The optimum CMCase activity monitored at different temperature levels for various bacillus strains was found to be lower than in this study (Abd Elhameed et al, 2020). The optimum temperature (60 °C) of the enzyme from B. subtilis CD001 was found to be comparably more than other bacterial strains and share similar characteristics when compared to some other strains of bacteria as well.…”

Section: Temperature and Ph Effectcontrasting

confidence: 60%

Biochemical Characterization of Cellulase From Bacillus subtilis Strain and its Effect on Digestibility and Structural Modifications of Lignocellulose Rich Biomass

Malik

Javed

2021

Front. Bioeng. Biotechnol.

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Microbial cellulases have become the mainstream biocatalysts due to their complex nature and widespread industrial applications. The present study reports the partial purification and characterization of cellulase from Bacillus subtilis CD001 and its application in biomass saccharification. Out of four different substrates, carboxymethyl cellulose, when amended as fermentation substrate, induced the highest cellulase production from B. subtilis CD001. The optimum activity of CMCase, FPase, and amylase was 2.4 U/ml, 1.5 U/ml, and 1.45 U/ml, respectively. The enzyme was partially purified by (NH4)2SO4 precipitation and sequenced through LC-MS/MS. The cellulase was found to be approximately 55 kDa by SDS-PAGE and capable of hydrolyzing cellulose, as confirmed by zymogram analysis. The enzyme was assigned an accession number AOR98335.1 and displayed 46% sequence homology with 14 peptide-spectrum matches having 12 unique peptide sequences. Characterization of the enzyme revealed it to be an acidothermophilic cellulase, having an optimum activity at pH 5 and a temperature of 60°C. Kinetic analysis of partially purified enzyme showed the Km and Vmax values of 0.996 mM and 1.647 U/ml, respectively. The enzyme activity was accelerated by ZnSO4, MnSO4, and MgSO4, whereas inhibited significantly by EDTA and moderately by β-mercaptoethanol and urea. Further, characterization of the enzyme saccharified sugarcane bagasse, wheat straw, and filter paper by SEM, ATR-FTIR, and XRD revealed efficient hydrolysis and structural modifications of cellulosic materials, indicating the potential industrial application of the B. subtilis CD001 cellulase. The findings demonstrated the potential suitability of cellulase from B. subtilis CD001 for use in current mainstream biomass conversion into fuels and other industrial processes.

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Section: Temperature and Ph Effectcontrasting

confidence: 60%

Biochemical Characterization of Cellulase From Bacillus subtilis Strain and its Effect on Digestibility and Structural Modifications of Lignocellulose Rich Biomass

Malik

Javed

2021

Front. Bioeng. Biotechnol.

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“…These results are in agreement with those reported for other Bacillus strains with high bioconversion potential. [25][26][27][28][29][30][31][32][33] After enzyme detection on agar plates and by zymography, eight nutritional factors including POW concentration, substrate particle size, NaCl concentration, POW liquor, Glucose concentration, bacterial strain, inoculum size and incubation time were selected in the present investigation as the key factors affecting the cellulase production by selected halotolerant bacteria. The levels of factors were selected by taking into account the operating experimental limits, and the previous studies.…”

Section: Resultsmentioning

confidence: 99%

Statistical optimization of cellulase production by halophilic strains isolated from Tunisian Sebkhas using the renewable marine biomass waste, Posidonia oceanica, as a cellulosic substrate

Souii¹,

Gorrab²,

Hammami³

et al. 2022

BBIJ

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Bio-based ethanol production from enzymatic lignocellulosic biomass degradation serves an efficient technology to combat the problem of depleting fossil fuels. High cost of the enzyme is the major obstacle in preventing the commercialization of this process. Thus, main objective of the present study was to optimize composition of culture parameters for enhancing cellulase production by two halophilic strains Oceanobacillus picture E50 and Bacillus vallismortis J77 using Posidonia oceanica waste (POW) as a substrate. A Plackett-Burman design was conducted to study and evaluate the weight of eight cultural variables on bacterial cellulase production. The optimal culture conditions for maximum enzyme yield were developed by maintaining the variables of POW concentration, NaCl concentration, POW liquor, bacterial strain and inoculum size at their higher levels, while keeping substrate particle size, glucose concentration and incubation time at their lower levels. The highest cellulase production in terms of CMCase activity was 950 U/L following this optimization process. Thus, the results obtained in this study demonstrated the potential of utilizing POW as a substrate for enhanced cellulase production by halophilic bacteria. B. vallismortis J77 produced the highest cellulase activity and therefore could be of great interest for saccharification of Posidonia oceanica biomass for biofuel and biorefinery applications.

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“…Diferentes períodos de incubação têm sido relatados para obter a melhor atividade celulolítica, como 48 h e 60 h para Bacillus sp. (Goyal V et al, 2014), 144 h para Geobacillus (Potprommanee L et al, 2017) ou mesmo períodos de incubação mais longos, como cinco dias de crescimento para alguns isolados de Bacillus (Abd Elhameed E et al, 2020).…”

Section: Resultsunclassified

“…(pH 7-8, a 37°C) (Mukhtar et al, 2019), Bacillus sp. (pH 7, a 40°C) (Elhameed et al, 2020) e B. thuringiensis (pH 8, a 60°C) (Awasthi MK et al, 2018), corroborando com os resultados obtidos no presente estudo. A celulases alcalinas podem ser adicionadas como aditivos detergentes ajudando a iluminar as cores, amaciar os tecidos e remover a sujeira; tais enzimas também funcionam para melhorar a resistência das formulações de polpa de papel nas indústrias de papel (Gupta et al, 2015).…”

Section: Drentensis Me-2)unclassified

Bioprospecção e caracterização de bactérias produtoras de celulase a partir do solo do Cerrado brasileiro

Alves

Rotta

Ferreira-Machado

et al. 2021

RSD

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Este trabalho descreve a bioprospecção e a caracterização de bactérias produtoras de celulase isoladas de solo obtido em uma região preservada do bioma Cerrado, em Uberaba, Brasil. A atividade celulolítica foi confirmada em 14 das 84 linhagens bacterianas isoladas. De acordo com a quantificação enzimática, cinco isolados foram selecionados como os melhores produtores de celulase e com base no sequenciamento parcial do gene 16S rRNA foram identificadas como Bacillus siamensis (isolado AB-9; 5.000U/mL), Bacillus toyonensis (isolado AB-1; 4.630U/mL), Bacillus methylotrophicus (isolados MB-3; 4.236U/mL e MP-7; 4.282U/mL) e Bacillus drentensis (isolado ME-2; 4.444U/mL). O extrato enzimático obtido de B. siamensis AB-9 foi o mais estável em diferentes valores de pH (2-8), mantendo sua atividade celulolítica, enquanto B. toyonensis AB-1 e B. methylotrophicus MP-7 produziram celulases com atividade máxima em pH 7 e 8. As celulases produzidas por B. siamensis AB-9 e B. toyonensis AB-1 apresentaram alta atividade enzimática em todas as temperaturas analisadas (10-80°C), enquanto as celulases de B. methylotrophicus MB-3 apresentaram atividade máxima na faixa de 20-70°C. Até o momento, esta é a primeira vez que bactérias produtoras de celulase isoladas do bioma Cerrado brasileiro na região do Triângulo Mineiro com potencial aplicação biotecnológica são descritas.

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Biochemical and Molecular Characterization of Five Bacillus Isolates Displaying Remarkable Carboxymethyl Cellulase Activities

Cited by 9 publications

References 33 publications

Biochemical Characterization of Cellulase From Bacillus subtilis Strain and its Effect on Digestibility and Structural Modifications of Lignocellulose Rich Biomass

Biochemical Characterization of Cellulase From Bacillus subtilis Strain and its Effect on Digestibility and Structural Modifications of Lignocellulose Rich Biomass

Statistical optimization of cellulase production by halophilic strains isolated from Tunisian Sebkhas using the renewable marine biomass waste, Posidonia oceanica, as a cellulosic substrate

Bioprospecção e caracterização de bactérias produtoras de celulase a partir do solo do Cerrado brasileiro

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