Lipases are enzymes that have the potential to hydrolyze triacylglycerol to free fatty acids and glycerol and have various applications. The aim of the present study was to isolate and screen marine bacteria for lipase production, optimize the production, and treat wastewater. A total of 20 marine bacterial isolates were obtained from the Mediterranean Sea and were screened for lipase production. All isolates were found to have lipolytic ability. The differences between the isolates were studied using RAPD-PCR. The most promising lipase producer (isolate 3) that exhibited the highest lipolytic hydrolysis (20 mm) was identified as Bacillus cereus HSS using 16S rDNA analysis and had the accession number MF581790. Optimization of lipase production was carried out using the Plackett–Burman experimental design with cotton seed oil as the inducer under shaking conditions at 10°C. The most significant factors that affected lipase production were FeSO4, KCl, and oil concentrations. By using the optimized culture conditions, the lipase activity increased by 1.8-fold compared with basal conditions. Immobilization by adsorption of cells on sponge and recycling raised lipase activity by 2.8-fold compared with free cells. The repeated reuse of the immobilized B. cereus HSS maintained reasonable lipase activity. A trial for the economic treatment of oily wastewater was carried out. Removal efficiencies of biological oxygen demand, total suspended solids, and oil and grease were 87.63, 90, and 94.7%, respectively, which is promising for future applications.
Seven morphologically distinct marine fungi were isolated from sediment and Seawater samples at different sites along Alexandria seashore. Antagonism effect against Aeromonas hydrophila on purpose and other pathogen was estimated. The most promising isolate giving the highest antibacterial activity (14 mm) against A. hydrophila was morphologically and genetically identified as Aspergillus terreus SHE05 and the corresponding sequence was recorded in the GenBank database with accession no. MW772239. Time course production of the antibacterial agents by A. terreus SHE05 against A. hydrophila was studied showing the highest productivity after 5 days incubation. Multi-factorial design in terms of Placket Burman design was implemented to predict the critical factors influencing the production of the antibacterial agents by A. terreus SHE05 against A. hydrophila. The obtained results showed that malt extract, pH and temperature were the key factors affecting the antimicrobial activity. Consequently, Box-Behnken design was applied to estimate the optimized levels of each independent variable showing that the optimized conditions were malt extract, 3 (g/l); peptone, 0.75 (g/l); salinity, 50%; pH, 4; culture age, 4 days; inoculum size, 0.5 ml; temperature 30°C and incubation time 5 days, which caused an increase in the antimicrobial activity to 25 mm, which denotes an approximately 1.8 fold increase comparing with the pre-optimized conditions. The potentiality of chloroform, hexane, petroleum ether and ethyl acetate for extraction of the active compounds was tested showing that ethyl acetate was the best. The extracted bioactive metabolites using ethyl acetate were tested as antimicrobial, anticancer, antiviral and antioxidant agents. Results showed reasonable activities. GC-MS was used to recognize the active components in the ethyl acetate extract, showing that the major compound was the Dodecanamine, N,N-Dimethyl with RT 11.95, molecular weight 213, area % (55.46) and molecular formula C14H31N.
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