Abdul Ghani Ghumro
Shah Abdul Latif University , Pakistan
Title: The Production of Extracellular Thermostable α-Amylase Enzyme from the Geobacillus stearothermophilus (BGSC 9A20) using soluble starch and optimization at various conditions.
Biography
Biography: Abdul Ghani Ghumro
Abstract
Microbial enzymes are for the most part utilized in modern procedures because of their minimal effort, large productivity, substance stability, environment protection, flexibility and huge availability. Some thermophilic Bacillus species utilized as bacterial workhorses in modern microbial developments for the creation of an assortment of enzymes and additionally fine biochemical for a considerable length of time. The α-Amylase are a standout amongst the most critical business compound about 25% to 33% of the world's enzyme market. Thus this study to isolate thermophiles bacteria from soil samples, samples were collected from Nara desert area (Khairpur Mir’s). Isolated bacterial culture (thermophilic Bacillus species) to inoculate on primary screening media for the detection of α-Amylase enzyme activity, Selected isolate was sent to Macrogen Inc., Seol, Korea, (as pure culture stocks) for commercial identification of the isolate through PCR amplification and sequencing of 16S rRNA gene. The effect of culture conditions on the production of α-Amylase enzyme, optimization parameters were optimized like temperature (45, 50, 55, 60 and 65°C), pH (6.0, 6.5, 7.0, 7.5, 8.0, 8.5 and 9.0 pH), agitation speed (100, 160, 200 and 250 rpm), age of inoculum (12, 24 and 36 hrs), size of inoculum (5, 10, 15 and 20%), under controlled condition. Therefore, it was observed that STB A1 isolate related with the reference strain (Geobacillus stearothermophilus (BGSC 9A20)), that produce a thermostable α-Amylase enzyme. Results were recorded by using the DNS assay and the protein estimation for detection of α-Amylase production and activity. So, the optimum temperature noted is 60°C (1639.78 U/ml), pH 7.5 (1734.44 U/ml), agitation speed 160 rpm (1745.18 U/ml), 24 hours age of inoculum (1750.11 U/ml) and 5% size of inoculum (1850.13 U/mi). Optimum enzyme activity in DNS assay condition, temperature stability at 60°C (1938.65 U/ml/min), reaction time is 35 min (2025.41 U/ml/min), dextrose substrate (2201.04 U/ml/min) and substrate concentration 1% (2326.11 U/ml/min), optimum conditions for the α-Amylase enzyme production and activity.