Bio Ethanol Production from Rice Straw Saccharification via Avicelase Gene in E. coli Recombinant Strain

Abdel-Salam, M. S.; Hafez, Safa S.; Fadel, Mohamed; Mohamed, Sahar S.; Hegazy, Wafaa K.; Khalil, Bigad E

doi:10.3390/cleantechnol5020023

Cited by 4 publications

(1 citation statement)

References 39 publications

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“…Numerous studies have employed the pGEM-Teasy vector for both cloning and heterologous gene expression. For example, Abdel-Salam et al [ 41 ] and their colleagues successfully cloned and expressed the avicelase Gene from Bacillus subtilis subsp. subtilis 168 in E. coli DH5α using the pGEM-Teasy vector.…”

Section: Discussionmentioning

confidence: 99%

Optimization, gene cloning, expression, and molecular docking insights for enhanced cellulase enzyme production by Bacillus amyloliquefaciens strain elh1

El-Khamisi,

Soliman,

El-Sayed

et al. 2024

Microb Cell Fact

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Background In this study, we isolated a cellulase-producing bacterium, Bacillus amyloliquefaciens strain elh, from rice peel. We employed two optimization methods to enhance the yield of cellulase. Firstly, we utilized a one-variable-at-a-time (OVAT) approach to evaluate the impact of individual physical and chemical parameters. Subsequently, we employed response surface methodology (RSM) to investigate the interactions among these factors. We heterologously expressed the cellulase encoding gene using a cloning vectorin E. coli DH5α. Moreover, we conducted in silico molecular docking analysis to analyze the interaction between cellulase and carboxymethyl cellulose as a substrate. Results The bacterial isolate eh1 exhibited an initial cellulase activity of 0.141 ± 0.077 U/ml when cultured in a specific medium, namely Basic Liquid Media (BLM), with rice peel as a substrate. This strain was identified as Bacillus amyloliquefaciens strain elh1 through 16S rRNA sequencing, assigned the accession number OR920278 in GenBank. The optimal incubation time was found to be 72 h of fermentation. Urea was identified as the most suitable nitrogen source, and dextrose as the optimal sugar, resulting in a production increase to 5.04 ± 0.120 U/ml. The peak activity of cellulase reached 14.04 ± 0.42 U/ml utilizing statistical optimization using Response Surface Methodology (RSM). This process comprised an initial screening utilizing the Plackett–Burman design and further refinement employing the BOX -Behnken Design. The gene responsible for cellulase production, egl, was effectively cloned and expressed in E. coli DH5α. The transformed cells exhibited a cellulase activity of 22.3 ± 0.24 U/ml. The egl gene sequence was deposited in GenBank with the accession number PP194445. In silico molecular docking revealed that the two hydroxyl groups of carboxymethyl cellulose bind to the residues of Glu169 inside the binding pocket of the CMCase. This interaction forms two hydrogen bonds, with an affinity score of −5.71. Conclusions Optimization of cultural conditions significantly enhances the yield of cellulase enzyme when compared to unoptimized culturing conditions. Additionally, heterologous expression of egl gene showed that the recombinant form of the cellulase is active and that a valid expression system can contribute to a better yield of the enzyme.

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Section: Discussionmentioning

confidence: 99%

Optimization, gene cloning, expression, and molecular docking insights for enhanced cellulase enzyme production by Bacillus amyloliquefaciens strain elh1

El-Khamisi,

Soliman,

El-Sayed

et al. 2024

Microb Cell Fact

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Bioethanol Production from Characterized Pre-treated Sugarcane Trash and Jatropha Agrowastes

Elnagdy,

Ragab,

Fadel

et al. 2024

Journal of Biotechnology

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A Study on the Valorization of Rice Straw into Different Value-Added Products and Biofuels

Ali,

Hosny,

Nassar

et al. 2024

International Journal of Chemical Engineering

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This work depicts that rice straw (RS), which is one of the major lignocellulosic wastes all over the world and causing many environmental problems, has considerable amounts of protein, ash, macronutrients, and micronutrients of approximately 11.38%, 16.77%, 2.27 mg/kg, and 771.9 mg/kg, respectively; besides, a C/N ratio of 15.18, a total N, P2O5, and K2O content of 1.85%, and a considerably low concentration of undesirable heavy metals and silica of approximately 77.69 mg/kg and 109 mg/kg are also present, which recommends its applicability as a precursor feedstock for the production of organic fertilizer and animal fodder. The batch solid-state fermentation (SSF) of RS by Trichoderma longibrachiatum DSMZ 16517 produced considerable amount of total reducing sugars (TRS) of approximately 339.2 mg TRS/g RS under the optimum operatic conditions of 20% (w:v) substrate concentration, pH 7, 1% inoculum size, a 9-day incubation period, and 30°C incubation temperature. The readily available and cost-effective agroindustrial waste, sugarcane molasses, proved to enhance the fungal biomass growth and (hemi) cellulolytic enzymes activities. The inoculated RS-SSF batch process with T. longibrachiatum precultured on 10% molasses enhanced the (hemi) cellulolytic enzymatic activities and TRS production rate by approximately 5.82 and 3.8 folds, respectively, relative to that inoculated by T. longibrachiatum precultured in the conventional potato dextrose broth medium. The separate hydrolysis and fermentation processes by different yeast strains Candida tropicalis DSM 70156, C. shehatae ATCC 58779, and Saccharomyces cerevisiae ATCC 64712 revealed an efficient bioethanol yield and productivity that ranged between 0.36 and 0.38 g/g sugars and 0.22 and 0.23 g/L/h, respectively, with concomitant competent fermentation efficiencies that ranged between 48.35% and 51.25%. The proximate analysis of rice straw before and after fungal hydrolysis proved calorific values of approximately 15.8 MJ/kg and 16.05 MJ/kg, respectively, recommending their applicability as primary and secondary solid biofuels. Thus, this study proved the waste prosperity of RS for environmental opulence and sustainability.

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Bio Ethanol Production from Rice Straw Saccharification via Avicelase Gene in E. coli Recombinant Strain

Cited by 4 publications

References 39 publications

Optimization, gene cloning, expression, and molecular docking insights for enhanced cellulase enzyme production by Bacillus amyloliquefaciens strain elh1

Optimization, gene cloning, expression, and molecular docking insights for enhanced cellulase enzyme production by Bacillus amyloliquefaciens strain elh1

Bioethanol Production from Characterized Pre-treated Sugarcane Trash and Jatropha Agrowastes

A Study on the Valorization of Rice Straw into Different Value-Added Products and Biofuels

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