Isolation and Phylogenetic study of new amylase producing Bacillus strain from Ghainarjeh hot spring in East Azarbaijan

Abstract

Background and objectives: Amylase is one of the important industrial enzymes, which is used in the food industry, detergent production, glucose, textile, and paper making. Bacillus spp. is considered as a major amylase producing microorganisms. The main objective of this study is isolation and phylogenic study of amylase producing thermophile Bacillus spp. from Ghainarjeh hot sprin in East Azarbaijan. Materials and methods: Samples from water of Ghainarjeh hot spring were cultured on recommended mediums. The grown colonies were tested for amylase production and positive colony determined a gram positive Bacillus using morphological and microscopically characteristics. Selected colony was used for total genomic DNA extraction, 16S rRNA fragment amplification and sequencing was done according to standard procedures. The sequence of 16S rRNA was used for phylogenetic investigation using maximum likelihood and maximum parsimony methods. Results: According to the results from morphological and biochemical investigations, the isolate is a gram positive, spore forming Bacillus with ability for use glucose, fructose and sucrose but not galactose and lactose. This isolate was positive for endol, citrate and VP but negative for catalase and oxidase. Conclusion: The results of phylogenetic investigation from both method show that the amylase producing isolated Bacillus is a new member of Bacillus Subtilis spp. in this study a new member of Bacillus subtilis spp. with the ability in amylase production was successfully isolated.

Keywords

• 16s rrna
• α-amylase
• phylogenetic
• ghainarjeh hot spring
• bacillus subtillis