Abstract
A newly isolated amylolytic strain was identified as Bacillus cereus spH1 based on 16S and 16-23S gene sequencing (Accession numbers OP811441.1 and OP819558, respectively), optimization strategies, using one variable at time (OVAT) and Plackett–Burman design, were employed to improve the alpha-amylase (α-amylase) production. Condition inferred revealed that the optimal physical parameters for maximum enzyme production were 30 °C, pH 7.5, and 12 h of incubation, using tryptone, malt extract, orange (Citrus sinensis) peels, crab (Portunus segnis) shells, calcium, and sodium chloride (NaCl) as culture medium. The full factorial design (FFD) model was observed to possess a predicted R2 and adjusted R2 values of 0.9788 and 0.9862, respectively, and it can effectively predict the response variables (p = 0). Following such efforts, α-amylase activity was increased 141.6-folds, ranging from 0.06 to 8.5 U/mL. The ideal temperature and pH for the crude enzyme activity were 65 °C and 7.5, respectively. The enzyme exhibited significant stability, with residual activity over 90% at 55 °C. The maltose was the only product generated during the starch hydrolysis. Moreover, the Bacillus cereus spH1 strain and its α-amylase were used in the treatment of effluents from the pasta industry. Germination index percentages of 143% and 139% were achieved when using the treated effluent with α-amylase and the strain, respectively. This work proposes the valorization of agro-industrial residues to improve enzyme production and to develop a green and sustainable approach that holds great promise for environmental and economic challenges.
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Data Availability
All the relevant data have been provided in the manuscript. Links to the submitted sequences: Bacillus cereus strain H1 16S ribosomal RNA gene, partial sequence: https://www.ncbi.nlm.nih.gov/nucleotide/OP811441.1?report=genbank&log$=nuclalign&blast_rank=1&RID=TZEBW479016; Bacillus cereus 16S-23S ribosomal RNA intergenic spacer and 23S ribosomal RNA gene, partial sequence: https://www.ncbi.nlm.nih.gov/nucleotide/OP819558.1?report=genbank&log$=nuclalign&blast_rank=1&RID=TZEPY6CJ013.
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Acknowledgements
This work is part of a doctoral thesis by Bouthaina Ben Hadj Hmida, whose research was financially supported by the Ministry of Higher Education and Scientific Research (Tunisia) through a grant to the Laboratory of Biochemistry and Enzymatic Engineering of Lipases-Engineering National School of Sfax- University of Sfax-Tunisia.
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BBHH: Data curation, Formal analysis, Investigation, Methodology, Writing—Original Draft. SBM and AF: Investigation, Methodology, Writing — Review & Editing. AS and AH-S: Conceptualization, Writing — Review & Editing, Supervision, Project administration.
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Ben Hadj Hmida, B., Ben Mabrouk, S., Fendri, A. et al. Optimization of newly isolated Bacillus cereus α-amylase production using orange peels and crab shells and application in wastewater treatment. 3 Biotech 14, 119 (2024). https://doi.org/10.1007/s13205-024-03962-3
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DOI: https://doi.org/10.1007/s13205-024-03962-3