Molecular characterization of gene encoding halo tolerant amylase of Bacillus paralicheniformis isolated from Jazan region

Abstract

Objectives: This study aims to characterize the gene encoding halo tolerant amylase of bacteria isolated from Jazan region. Materials and Methods: Soil samples were collected from several area of Jazan region, KSA. The samples were serially diluted and plateted on starch agar plates. The amylase producing bacteria were detected by iodine test. To determine the halophilic amylase producing bacteria, several colonies were tested for their ability to grow at higher concentrations of NaCl ranging from 7 to 16%. The bacteria was identified by 16S rRNA and the full length amylase gene was fully identified by sequencing using specific primers. Results: One bacterial halophilic isolate was able to grow on starch agar medium up to 14% NaCl. The Gram stain of the isolate indicated that it is Gram-positive, bacilli. The 16S rRNA gene homology study showed that the bacterial isolate was identified as Bacillus paralicheniformis. Two specific primers were designed named S1F, S1R, to amplify the amylase gene (AMY) region using PCR and the PCR product was sequenced. The sequencing results showed that the full-length amy gene of B. paralicheniformis was of 1452 encoding 483 amino acids. The expected M.Wt. of the protein expressed is of 55 KDa. Conclusion: We report the isolation, identification, and characterization of an isolate of halophilic bacterium isolated from Jazan region. Based on molecular identification, this isolate was identified as Bacillus paralicheniformis. This bacterial strain has an α-amylase gene in its genome and is able to produce extracellular α-amylase. Based on the findings of this work we propose that Bacillus paralicheniformis amy gene could be cloned into expression vector for large scale production.

Diversity and activity of amylase-producing bacteria isolated from mangrove soil in Thailand

“…Bacillus paralicheniformis was isolated from a fermented soybean paste (Cheonggukjang), identified, and characterized in 2015 (Dunlap et al 2015). This bacterium was reported to have an -amylase gene in its genome and was able to produce extracellular -amylase (Ghazouani et al 2020). The amylases from B. paralicheniformis were proven to be a highly efficient raw starch digesting enzyme (Božić et al 2020).…”

“…The amylases from the B. paralicheniformis strain DNP0507 could be possible as -amylases because theses enzymes were usually produced by bacteria belonging to the genus Bacillus (Sajedi et al 2005). Moreover, the studies of Bekler et al (2019), Božić et al (2020), andGhazouani et al (2020) have identified the amylases from B. paralicheniformis as -amylases. However, the identification of the B. paralicheniformis strain DNP0507 amylases should be further verified by molecular weight and amino acid sequencing analyses.…”

Biosynthesis of silver nanoparticles by Nocardiopsis sp.‐MW279108 and its antimicrobial activity

“…al. [9]. The PCR product was electrophorized on low melting agarose (1%) and purified using the PCR purification Kit (Thermo).…”