Abstract
Enzymes are proteins that accelerate the rate of various biochemical reactions in living organisms without being used up in the reaction. Enzymes are involved in most biochemical reactions occurring in microorganisms, plants, animals, and human beings. Enzymes are widely used to promote industrial processes and product manufacturing; hence sometimes called industrial enzymes. Hydrolytic enzymes represent one of the most diverse categories of microbial enzymes and are of special interest to researchers. Despite the extensive history of its research, this group of enzymes continues to exhibit excellent potential for use in the medicinal, food, biofuels, and agricultural industries. Therefore, the distinctive qualities of Bacillus enzyme such as stability in a variety of temperatures and pH, high specificity, the biodegradability of a wide range of substrates and genetic manipulation, offer a sufficient basis for the development of innovative biotechnologies. This study explored the production of enzymes by Bacillus paranthracis strain MHSD3 isolated from Pellaea calomelanos. Enzyme production was screened through the agar plate method and different parameters to enhance enzyme production were bio-prospected through submerged fermentation. Bacillus paranthracis strain MHSDMHSD3 tested positive on the following enzymes: amylase, L-asparaginase, cellulase, chitinase, lipase, and pectinase. This study displayed enzyme activities of amylase (31788.59 IU), cellulase (4487.486 IU), chitinase (6.068182 IU), L-asparaginase (12.8912 IU), lipase (16.3873 IU) and pectinase (13.98986 IU). The antioxidant and anti-inflammatory activity of enzymes Bacillus paranthracis MHSD3, a probiotic candidate isolated from Pellaea calomelanos, was investigated using in vitro models such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and protein denaturation. Enzyme activity from probiotic strain B. paranthracis was able to stabilize the DPPH and ABTS to its neutral form. The obtained DPPH radical scavenging activity of the enzymes were as follows: amylase (62.21±.0.057%), cellulase (58.37±0.51%), chitinase (75.26±0.06%), L-asparaginase (60.58±0.05%), lipase (75.78±0.19%), and pectinase (55.86±0.06%). The ABTS scavenging activity of the enzymes were amylase (52.53±0.08%), cellulase (59.16±0.08%), chitinase (64.27±4.34%), L-asparaginase (68.17±0.22%), lipase (71.87±0.30%) and pectinase (51.51±0.82%). The in vitro protein denaturation assay was used to measure the anti-inflammatory activity of enzymes from Bacillus paranthracis strain MHSD3. L-asparaginase had the highest activity (73.18±0.15%), while the other enzymes had the least amount of activity (chitinase, 53.27±56%) and cellulase, 49.77±0.54%). The results suggested that all enzymes from B. paranthracis strain MHSD3 have compelling antioxidant properties with L-asparaginase, chitinase and cellulase effectively protected from denaturation.