Bioprocess Optimization for Enhanced Production of Bacterial Cellulase and Hydrolysis of Sugarcane Bagasse

Malik, Waseem Ayoub; Khan, Haris M.; Javed, Saleem

doi:10.1007/s12155-021-10259-3

Cited by 19 publications

(18 citation statements)

References 49 publications

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“…"Bacillus subtilis", A potential cellulase producing bacteria was successfully isolated from cow dung as described earlier (Malik et al, 2021). The nucleotide sequence was deposited to NCBI GeneBank and an accession number MN173007 was obtained.…”

Section: Microorganism Reagents and Preparations Of Lignocellulosic Materialsmentioning

confidence: 99%

“…The nucleotide sequence was deposited to NCBI GeneBank and an accession number MN173007 was obtained. It is also preserved at CSIR-Institute of Microbial Technology, Chandigarh (India), under deposit number MTCC12940 (Malik et al, 2021). The bacteria was routinely cultured on CMC agar medium, sub-culturized at regular intervals, and kept at 4 °C during their storage time.…”

Section: Microorganism Reagents and Preparations Of Lignocellulosic Materialsmentioning

confidence: 99%

“…Bacterial species, such as B. subtilis subsp. subtilis JBS250 (Kumar and Singh, 2021), B. subtilis strain LFS3 (Rawat and Tewari, 2012), B. subtilis Strain CBS31 (Sudip et al, 2020), Bacillus halodurans IND18 (Vijayaraghavan et al, 2016), B. subtilis BC1 (Dehghanikhah et al, 2020), B. subtilis BY-4 (Ma et al, 2015), and B. subtilis CD001 (Malik et al, 2021) have been recently reported for cellulase production. Bacteria reveal a wide variety of physicochemical conditions.…”

Section: Introductionmentioning

confidence: 99%

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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: Microorganism Reagents and Preparations Of Lignocellulosic Materialsmentioning

confidence: 99%

Section: Microorganism Reagents and Preparations Of Lignocellulosic Materialsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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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“…As represented by B. subtilis, a lot of studies regarding cellulase gene sequences, enzymatic activities, optimal condition for cellulolysis were published from Bacillus species. Cellulases from Bacillus are still now reported frequently [10,11].…”

Section: Introductionmentioning

confidence: 99%

Genome sequencing and identification of cellulase genes in Bacillus paralicheniformis strains from the Red Sea

et al. 2021

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Background Cellulolytic microorganisms are considered a key player in the degradation of plant biomass in various environments. These microorganisms can be isolated from various environments, such as soils, the insect gut, the mammalian rumen and oceans. The Red Sea exhibits a unique environment in terms of presenting a high seawater temperature, high salinity, low nutrient levels and high biodiversity. However, there is little information regarding cellulase genes in the Red Sea environment. This study aimed to examine whether the Red Sea can be a resource for the bioprospecting of microbial cellulases by isolating cellulase-producing microorganisms from the Red Sea environment and characterizing cellulase genes. Results Three bacterial strains were successfully isolated from the plankton fraction and the surface of seagrass. The isolated strains were identified as Bacillus paralicheniformis and showed strong cellulase activity. These results suggested that these three isolates secreted active cellulases. By whole genome sequencing, we found 10 cellulase genes from the three isolates. We compared the expression of these cellulase genes under cellulase-inducing and non-inducing conditions and found that most of the cellulase genes were generally upregulated during cellulolysis in the isolates. Our operon structure analysis also showed that cellulase genes form operons with genes involved in various kinds of cellular reactions, such as protein metabolism, which suggests the existence of crosstalk between cellulolysis and other metabolic pathways in the bacterial isolates. These results suggest that multiple cellulases are playing important roles in cellulolysis. Conclusions Our study reports the isolation and characterization of cellulase-producing bacteria from the Red Sea. Our whole-genome sequencing classified our three isolates as Bacillus paralicheniformis, and we revealed the presence of ten cellulase orthologues in each of three isolates’ genomes. Our comparative expression analysis also identified that most of the cellulase genes were upregulated under the inducing conditions in general. Although cellulases have been roughly classified into three enzyme groups of beta-glucosidase, endo-β-1,4-glucanase and exoglucanase, these findings suggest the importance to consider microbial cellulolysis as a more complex reaction with various kinds of cellulase enzymes.

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“…In some processes, lignin is first broken down and removed by special enzymes or chemicals (such as alkali), before further steps are taken to convert the remaining part of the SCB to bio-ethanol (Niju and Swathika 2019). (Maryana et al 2014) Lignin peroxidase, manganese peroxidase and laccase are the major enzymes for de-lignification and can be produced by white-rot fungi (Malik et al 2021). Asgher et al (2013) undertook a comparison of both enzymatic and alkali de-lignification of SCB.…”

Section: Preparation Strategiesmentioning

confidence: 99%

Sugarcane bagasse: a biomass sufficiently applied for improving global energy, environment and economic sustainability

Ajala

Ighalo

Ajala

et al. 2021

Bioresour. Bioprocess.

124

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Sugarcane (Saccharum officinarum) bagasse (SCB) is a biomass of agricultural waste obtained from sugarcane processing that has been found in abundance globally. Due to its abundance in nature, researchers have been harnessing this biomass for numerous applications such as in energy and environmental sustainability. However, before it could be optimally utilised, it has to be pre-treated using available methods. Different pre-treatment methods were reviewed for SCB, both alkaline and alkali–acid process reveal efficient and successful approaches for obtaining higher glucose production from hydrolysis. Procedures for hydrolysis were evaluated, and results indicate that pre-treated SCB was susceptible to acid and enzymatic hydrolysis as > 80% glucose yield was obtained in both cases. The SCB could achieve a bio-ethanol (a biofuel) yield of > 0.2 g/g at optimal conditions and xylitol (a bio-product) yield at > 0.4 g/g in most cases. Thermochemical processing of SCB also gave excellent biofuel yields. The plethora of products obtained in this regard have been catalogued and elucidated extensively. As found in this study, the SCB could be used in diverse applications such as adsorbent, ion exchange resin, briquettes, ceramics, concrete, cement and polymer composites. Consequently, the SCB is a biomass with great potential to meet global energy demand and encourage environmental sustainability.

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Bioprocess Optimization for Enhanced Production of Bacterial Cellulase and Hydrolysis of Sugarcane Bagasse

Cited by 19 publications

References 49 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

Genome sequencing and identification of cellulase genes in Bacillus paralicheniformis strains from the Red Sea

Sugarcane bagasse: a biomass sufficiently applied for improving global energy, environment and economic sustainability

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