Abstract
The bacterial strain producing thermostable, alklophilic alpha-amylase was identified as Bacillus amyloliquefaciens KCP2 using 16S rDNA gene sequencing data (NCBI Accession No: KF112071). Medium components were optimized through the statistical approach for the synthesis of alpha-amylase by the organism under solid-state fermentation using wheat bran as the substrate. The medium components influencing the enzyme production were identified using a two-level fractional factorial Plackett–Burman design. Among the various variables screened, starch, ammonium sulphate and calcium chloride were found to be most significant medium components. The optimum levels of these significant parameters were determined employing the response surface Central Composite design which significantly increased the enzyme production with the supplementation of starch 0.01 g, ammonium sulphate 0.2 g and 5 mM calcium chloride in the production medium. Temperature and pH stability of the alpha-amylase suggested its wide application in the food and pharmaceutical industries.
Highlights
Amylases find potential application in number of industrial processes such as bread making, brewing, starch processing, pharmacy, textile industries
The growth and the enzyme production by the organisms are strongly influenced by medium composition, and optimization of the medium components may lead to improved enzyme productivity (Djekrif et al 2006)
A 800-bp size 16S rDNA sequence of the isolate was obtained through PCR amplification and sequencing
Summary
Amylases find potential application in number of industrial processes such as bread making, brewing, starch processing, pharmacy, textile industries. Alpha-amylase is an extracellular enzyme that randomly cleaves the 1,4 a-Dglucosidic linkages between adjacent glucose units in the linear amylose chain. It is secreted as primary metabolite of microorganisms and its production is a growth-related process (Kammoun et al 2008). To meet the growing demands of amylase for industrial application, it is necessary to produce the highly efficient enzymes at large scale with reduced production cost (Haq et al 2003). Use of agro-industrial residues as the substrate for the fermentation has growing interests as they are inexpensive energy-rich resources and eliminates large-scale accumulation of the biomass (Pandey et al 2000; Ramachandran et al 2007). The growth and the enzyme production by the organisms are strongly influenced by medium composition, and optimization of the medium components may lead to improved enzyme productivity (Djekrif et al 2006)
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