Abstract
A natural bacterial strain identified as Bacillus amyloliquefaciens MBAA3 using 16S rDNA partial genome sequencing has been studied for optimization of cellulase production. Statistical screening of media components for production of cellulase by B. amyloliquefaciens MBAA3 was carried out by Plackett–Burman design. Plackett–Burman design showed CMC, MgSO4 and pH as significant components influencing the cellulase production from the media components screened by Plackett-Burman fractional factorial design. The optimum concentrations of these significant parameters were determined employing the response surface central composite design, involving three factors and five levels was adopted to acquire the best medium for the production of cellulase enzyme revealed concentration of CMC (1.84 g), MgSO4 (0.275 g), and pH (8.5) in media for highest enzyme production. Response surface counter plots revealed that middle level of MgSO4 and middle level of CMC, higher level of CMC and lower level of pH and higher level of MgSO4 with lower level of pH increase the production of cellulase. After optimization cellulase activity increased by 6.81 fold. Presence of cellulase gene in MBAA3 was conformed by the amplification of genomic DNA of MBAA3. A PCR product of cellulase gene of 1500 bp was successfully amplified. The amplified gene was conformed by sequencing the amplified product and sequence was deposited in the gene bank under the accession number KF929416.Graphical Response surface graph showing interaction effects between concentration of a CMC and MgSO4. b pH and CMC. c MgSO4 and pH
Highlights
Cellulases (1,4-(1,3;1,4)-b-D-glucan 4-glucanohydrolase) refers to a group of enzymes which hydrolyze cellulose
A natural bacterial strain identified as Bacillus amyloliquefaciens MBAA3 using 16S rDNA partial genome sequencing has been studied for optimization of cellulase production
Plackett–Burman design showed CMC, MgSO4 and pH as significant components influencing the cellulase production from the media components screened by Plackett-Burman fractional factorial design
Summary
Cellulases (1,4-(1,3;1,4)-b-D-glucan 4-glucanohydrolase) refers to a group of enzymes which hydrolyze cellulose. Cellulose is the most abundant organic source of food, fuel and chemicals [1]. It is the primary product of photosynthesis in terrestrial environments and the most abundant renewable bioresource produced in the biosphere [2]. Productivity of microbial enzymes and metabolites can be increased by manipulating nutritional requirements, physical parameters and genetic make up of the producing strain. RSM is considered as a standard statistical approach designing experiments, building models, evaluating the effects of many factors and finding the optimal conditions for desirable responses and reducing the number of required experiments [10]. The role of each variable, their interactions and statistical analysis for cellulase production were explained by applying the second-order polynomial model. This study describes the identification of gene encoding for this enzyme and the determination of its nucleotide sequence
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