Production, Optimization and Partial Characterization of Thermostable and Alkaline Amylase from Bacillus licheniformis KSU-6

Abstract

Total of 56 bacterial isolates were isolated from five palm tree cultivated soil samples and screened their amylase production. In the preliminary screening, twenty four isolates exhibited amylase producing ability. Among these 24 isolates, an isolate KSU-6 showed maximum amylase enzyme production (37.52 U mL -1 ), and the isolate was identified as Bacillus licheniformis KSU-6. The maximum amylase production was optimized, and the conditions were pH 10.0, temperature 50 o C, incubation period 44 h, inoculum concentration 2% (v/v), starch 0.2% (w/v) as carbon source, peptone 0.5% (w/v) along with malt extract 0.3% (w/v) as nitrogen sources for B. licheniformis KSU-6. The partially purified amylase stabled upto pH 12, temperature 80 o C, 1-2 M salt concentration and 10 mM of different detergents. In this study conclude that the soil bacterium Bacillus licheniformis KSU-6 produced thermostable and alkaline amylase, which could be useful for industrial liquefaction and detergent industries.

Purification, characterization, and statistical optimization of a thermostable α-amylase from desert actinobacterium Streptomyces fragilis DA7-7

“…Narayana and Vijayalakshmi (2008) reported that the optimum temperature for a-amylase production in Streptomyces albidoflavus at 30°C. Another recent study also confirmed the optimum aamylase production by S. cheonanensis VUK-A at 30°C (Naragani et al 2015).The incubation period is one of the major factors that influence a-amylase production (Dhanasekaran et al 2006;Nithya et al 2016). In this study, the optimum incubation period for a-amylase production by S. fragilis DA7-7 was found to be 72 h. In the previous report, Naragani et al (2015) also supported that the average incubation period employed was 72 h for a-amylase production by S. cheonanensis VUK-A.…”

“…Then, the precipitate was disbanded in 0.05 M glycine-NaOH buffer (pH 10). The mixture was dialyzed overnight by glycine-NaOH buffer (Nithya et al 2016).…”

“…The pH stability of purified amylase was studied by preincubating the enzyme (80 lg/mL) in different pH value buffers (4-12), at 37°C for 1 h. Thermal stability of the enzyme was determined from 40 to 100°C for 1 h. Similarly, halostability of the enzyme was tested different concentrations of NaCl (0-4 M) at 37°C for 1 h. The effect of various detergents (10 mM each) was tested at 37°C for 30 min: cetyltrimethylammonium bromide, sodium dodecyl sulfate, Triton X-100, Tween 20, and Tween 80 (Nithya et al 2016). The enzyme activities were determined and compared to the control.…”

Statistical-Based Bioprocess Optimization of Amylase Production from Halophilic Bacillus sp. H7.

“…Similarly, Mishra et al [30] and Nithya et al [31] have been reported that 1.5% of inoculum concentration was optimum for maximum amylase production for Bacillus sp. and Bacillus licheniformis KSU-6 respectively.…”

Statistical Based Bioprocess Design for Improved Production of Amylase from Halophilic Bacillus sp. H7 Isolated from Marine Water

“…The production will be affected by high concentration due to the increase in moisture contains as well as biomass production, whereas the low concentration results in the prolonged production period to transform the substrate into the product [32]. Similarly, Mishra et al [33] and Nithya et al [34] have reported that 1.5% of inoculum concentration was optimum for maximum amylase production for Bacillus sp. and Bacillus licheniformis KSU-6, respectively.…”