Biochemical and Molecular Characterization of Five Bacillus Isolates Displaying Remarkable Carboxymethyl Cellulase Activ
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Biochemical and Molecular Characterization of Five Bacillus Isolates Displaying Remarkable Carboxymethyl Cellulase Activities Esraa Abd Elhameed1 · Alaa R. M. Sayed1,4 · Tharwat E. E. Radwan2 · Gamal Hassan3 Received: 23 November 2019 / Accepted: 15 July 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Cellulases have many useful applications in industry and biotechnology. So, identification of new bacterial strains expressing cellulases with better properties is desired. Five soil bacterial strains screened for high carboxymethyl cellulase (CMCase) activities were characterized and identified by 16S rRNA analysis as Bacillus amyloliquefaciens (FAY088), B. velezensis (FAY0103), B. tequilensis (FAY0117), B. subtilis (FAY0136), and B. subtilis (FAY0182). Their CMCase activities were 1.49, 1.26, 1.21, 1.21, and 1.24 U/ml, respectively. The maximum CMCase production was attained by growth at 35 °C, pH 6, and 180 rpm for 5 days. Residual activities of CMCases from FAY088 and FAY0117 were 88% or more after growth at 40 °C, which is same as FAY0182 CMCase at 40 and 45 °C. Additionally, FAY0182 retained 73% residual activity at 50 °C. FAY088 and FAY0182 retained more than 85% at pH 7 and 8. Conversely, residual activities from FAY0103 and FAY0136 declined a lot by increasing growth temperature beyond 40 °C and pH beyond 7. The maximum CMCase stability in all isolates was observed at pH 7, 3-h incubation, and 40 °C except for FAY0103 CMCase showed optimum temperature at 30 °C. More than 70% CMCase stability was retained in case of FAY088 at 50 °C, FAY0117 at 50–70 °C, and FAY0136 at 50–60 °C. FAY088 CMCase seemed to be the lest sensitive to temperature variation as it displayed residual activities 67, 72, 78, 84, 77, 74, and 72% at pH 3, 4, 5, 6, 8, 9, and 10, respectively. Finally, the five CMCase-producing isolates are recommended further enzyme applications in biotechnology and industry.
Introduction Cellulases are used in many applications like food processing, agriculture, animal feed, biofuel, wine, textile, paper, pulp, laundry, and pharmaceutical industries [1–3]. Also, Esraa Abd Elhameed and Alaa R. M. Sayed have contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00284-020-02135-8) contains supplementary material, which is available to authorized users. * Alaa R. M. Sayed [email protected]; [email protected] 1
Chemistry Department, Faculty of Science, Fayoum University, Fayoum 63514, Egypt
2
Botany Department, Faculty of Science, Fayoum University, Fayoum 63514, Egypt
3
Genetics Department, Faculty of Agriculture, Fayoum University, Fayoum 63514, Egypt
4
College of Pharmacy, University of Florida, Orlando, FL 32827, USA
it can be used in waste and pollution management through degradation of agricultural and food wastes containing cellulosic residues by converting these materials into valuable products like glucose. The use of some biological agents like nonpathogenic bacteria
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