BIOPROSPECTING FOOD INDUSTRY EFFLUENTS FOR ENZYME-PRODUCING BACTERIA: MOLECULAR CHARACTERIZATION AND PRODUCTION ENHANCEMENT

Abstract

Industrial effluents are known reservoirs of diverse microorganisms with valuable biotechnological potential. There has not been much research done in Pakistan on using bacteria from wastewater to produce enzymes. In this work, food industry effluents were collected using the direct sampling (DS) method at the Hayatabad Industrial Estate in Peshawar in order to identify, filter, and optimize bacterial strains that produce industrially significant enzymes. Out of the 23 bacterial isolates that were collected, hydrolytic zone formation demonstrated that 12 (52.17%) had protease activity, 7 (31.13%) had cellulase activity, and 6 (26.08%) had pectinase activity. Both Gram-positive and Gram-negative isolates were found by microscopic and Gram staining, and biochemical analysis showed that Bacillus cereus, Pseudomonas granadensis, and Enterobacter cloacae were strong makers of enzymes. 16S rRNA sequencing was used to establish the molecular identity, and phylogenetic analysis revealed 99% similarity to reference strains. According to optimization experiments, Pseudomonas granadensis displayed peak cellulase activity at 45 °C (0.41 absorbance), Bacillus cereus reached maximum protease activity at 37 °C (0.51 absorbance), and Enterobacter cloacae displayed optimal pectinase activity at 30 °C (0.41 absorbance). At neutral pH 7.0, all isolates showed the highest enzyme yields; production peaked after 48 hours of incubation and then began to decline. Citrus pectin increased pectinase activity (0.39 absorbance), starch modestly supported cellulase synthesis (0.26 absorbance), and glucose was the most efficient carbon source for protease generation (0.52 absorbance) among the substrates examined. Peptone facilitated the highest levels of cellulase (0.39) and protease (0.51) activity among nitrogen sources, but yeast extract enhanced the formation of pectinase (0.42). These results demonstrate the encouraging potential of bacterial isolates from wastewater as long-term sources of industrial enzymes when cultured under ideal circumstances.