Aims: The objective of the study was to produce and optimize protease and pectinase from Bacillus subtilis isolated from market waste. Place and Duration of Study: Department of Microbiology (laboratory unit), Michael Okpara University of Agriculture Umudike, Abia State Nigeria. Methodology: The production and optimization of protease and pectinase from bacteria isolated from solid market waste was investigated. Isolated bacteria from the waste were screened for protease and pectinase production using skim milk agar and pectin agar respectively. Using morphological, biochemical and molecular technique the enzymes producing isolate was confirmed as Bacillus subtilis. Protease and Pectinase were produced by Bacillus subtilis using submerged fermentation in gelatin broth and pectin broth respectively. The enzymes were purified using ammonium sulphate precipitation, dialysis and ion-exchange chromatography. Optimization using different temperatures, pH and nutrient sources was done. Enzyme activity was measured. Results: Purified protease exhibited maximum activity of 8.72U/ml at 40oC while pectinase exhibited maximum activity of 8.98U/ml at 50oC. Glucose as a carbon source and peptone as a nitrogen source gave optimum activity for both enzymes. Both pectinase and protease exhibited optimum activity at pH 9. There was significant difference (P=.05) in enzyme activity at different temperatures, pH and nitrogen sources for both protease and pectinase. There was no significant difference in pectinase activity at P=.05 for the different carbon sources while there was significant difference for protease activity for the different carbon sources at P=.05. Conclusion: Production of microbial enzymes such as protease and pectinase from waste material is an eco-friendly process and cheaper option for large scale use of enzymes in industry.
The bio-scouring of cotton using protease and pectinase produced from Bacillus subtilis was investigated. Protease and pectinase were produced from Bacillus subtilis in a liquid medium using the submerged fermentation technique. Both enzymes were purified, and their scouring potential was tested on raw cotton fabrics. Pectinase was more effective than protease under optimised conditions. The optimum scouring temperature for both enzymes was between 40 °C and 50 °C, with pectinase bio-scoured fabric showing 15.5% weight loss while protease bio-scoured fabric had 14.3% weight loss. The optimum pH for pectinase scouring was pH 9 with 14.8% weight loss in the fabric, while the optimum pH for protease scoured fabric was pH 7 with 12.3% weight loss in fabric. After 120 minutes of bio-scouring, maximum weight loss was recorded for both pectinase and protease treated fabrics. The application of protease and pectinase for cotton fabric scouring revealed that they could be used as bio-scouring agents to treat textile materials.
Aims: The objective of the study was to ascertain the antimicrobial susceptibility pattern and ESBL prevalence of bacteria isolated from snacks. Place and Duration of Study: Department of Microbiology (Laboratory Unit) Michael Okpara University of Agriculture Umudike. Methodology: The snacks were mashed aseptically, serially diluted and inoculated onto nutrient agar and MacConkey agar. Isolates were identified using standard microbiological procedures. Antimicrobial susceptibility of the isolates and ESBL detection was done using disk diffusion method. ESBL production was confirmed using Double Disc Synergy Test (DDST) method following CLSI recommendations. Results: Escherichia coli, Salmonella Typhi, Staphylococcus aureus and Klebsiella pneumoniae were the bacteria isolated with Escherichia coli as the most prevalent isolate with 42% occurrence in the samples screened. There was significant difference in the sensitivity of the bacteria isolates to the different antibiotics used at P=0.05. Salmonella Typhi isolates exhibited highest resistance to an antibiotic with 86% resistance to ciprofloxacin while Klebsiella pneumoniae isolates exhibited the lowest resistance to an antibiotic with 10% resistance to cefotaxime. Among the Gram-negative bacteria, 36% of suspected ESBL producing E. coli isolates were confirmed as ESBL producers indicating the highest occurrence. Conclusion: The study confirmed the presence of bacteria in street vended snacks which exhibited high resistance to antibiotics that could be attributed to the presence of ESBL producers among the isolates.
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