Biotechnological application of bacterial alkaline thermostable enzymes in bio-detergent industry

This is an investigation concerned on the production of alkaline thermostable microbial enzymes for application in biodetergent technology. Bacillus licheniformis- B42 and Geobacillus stearothermophilus -B78 were selected and identified among one hundred and fifty-three thermophilic bacterial isolates with respect to their ability to produce alpha-amylases, cellulases, proteases and lipases grown on some agro-industrial wastes at 55 degree C and at pH 9 for application in biodetergent technology. Productivity of four alkaline thermostable enzymes by both selected strains using slaughter house wastes [SHW] as best substrate for proteases and lipases and potato peel [PP] as best substrate for alpha-amylases and cellulases. The enzymatic level more affected by incubation temperature, pH, SHW and PP concentrations, inoculum size, incubation period, carbon, nitrogen, metal inducer and vitamins sources, under shaking conditions. Four alkaline thermostable enzymes were produced under all optimal nutritional and environmental conditions and purification by column chromatography on Sephadex G200 and G100, respectively were performed. Purification of four produced alkaline thermostable enzymes steps resulted in raising the purification fold to 17.04,15.24, 411.9 and 27.33 times in comparable with crude enzymes for alpha-amylase. cellulase, protease and lipase, respectively. The wash performing analysis of the four enzymes revealed that, it could effectively remove a variety of stains such as blood, apple, chocolate, mango, strawberry, salad and pomegranate by treatment at 55 degree C for 15 min when alkaliphilic-thermostable crude/purified enzymes were added separately or in combination with or without detergent [Rabso] as an Egyptian local detergent product. The crude enzymes of these two bacterial strains proved to be potentially candidates for the application in the detergent technology

Relation between plasma and milk-clotting enzymes with proteolytic and fibrinolytic activities of some fungi

Extracellular plasma and milk clotting as well as proteolytic and fibrinolytic activities of 9 fungal species were investigated under different culturing conditions. There is an inverse relationship between the clotting activity and pH and dry mycelial weight. The clotting activity increased markedly at the acidic conditions. The clotting activity varied with the type of growth media and with the fungal species. No regular relationship could be detected between the proteolytic, fibrinolytic and clotting activities in different fungi grown on different media. On the other hand, no relation was observed between the clotting activity of either milk or plasma indicating that they are due to different enzymes. The milk clotting, proteolytic ratio C/P showed a great variation among fungi and growth media. The same was held true in the plasma- clotting-fibrinolytic C/F activity ratio. Aspergillus sydowi was the most promising clotting fungus either for milk or plasma and characterized by low proteolytic and fibrinolytic activities when grown on blood medium. For milk dotting activity only, Mucor pusillus grown on Stefanini medium recorded potent C/P ratio. For plasma clotting only, the following species recorded high C/F ratios, Aspergillus flavus, A. fumigatus and Cladosporinum flavum on Stefanini medium. A. niger on casein medium and A. niger, Seopulariopsis brumbtii and Cladosporinum flavum on blood medium

Plasma, blood and milk-clotting enzymes of some fungi

Isolation of fungal species from blood and whey by centrifugation technique was earned out. Aspergillus niger was the most frequent species in both sources. A. sydowi, A. tubingensis, Candida albicans and Cladosporium flavum dominated the blood cultures while Candida albicans, C. tropicalis, Cladosporium flavum, Aspergillus sydowi and Paecilomyces divaricated dominated whey cultures. The isolated fungi were mesophilic, thermophilic or thermotolerant. Plasma, blood and milk-clotting assays revealed that extracellular enzymes in the culture filtrates were more active in clotting the three substrates than intracellular enzymes in cell-free extract The whole blood clotted faster than its plasma. The results also showed that extracellular and intracellular enzymes were different and their activity depended upon the age of culture and kind of fungal species. Among nine assayed species, Aspergillus sydowi was the most promising in clotting whole blood, plasma and milk either extracellularly or intracellularly

Whey utilization by saccharomyces cerevisiae for production of biomass protein

Fayoum cheese whey from buffalo milk at different concentrations was used for growth and protein production by Saccharomyces cerevisiae. Analysis of whey sample indicated that it contained 88.7 percent water 11.3 percent total solids, 2.6 percent ash. The total reducing sugars, total nitrogen and total soluble salts constituted 18.3 percent, 16.2 percent and 3.9 percent respectively. The protein content represented 31.6 percent of the whey sample. Media containing whey, at all concentrations, recorded higher growth with better fermentation yield than control medium. Among the different concentration tested, 5 percent whey containing medium was the most favorable for production of maximum biomass protein. The suitable fermentation period was 4 days. Analysis of the harvested yeast cells on the tested media indicated also that the yeast grown on 5 percent whey containing medium has the highest percentage of protein [55.6 percent of the dry biomass] and promising amount of total sterols. The protein conversion coefficient revealed that 50.5 percent of the utilized sugar by S. cerevisiae was converted into protein

Ureolytic activity and growth criteria of some soil fungi affected by various concentrations of urea

It was found that there is no regular relation between the density of fungi isolated from soil amended with urea and their ureolytic activity. For instance, Aspergillus fumigatus constituted the highest density and ureolytic activity but Fusarium nivale was of low density and high ureolytic activity. Oppositely, high density but showed weak ureolytic activity. The effect of various urea concentrations added to culture medium as sole N-source showed that the ED50 was 8.5 percent urea in Aspergillus fumigatus, 6.5 percent in A niger, A. Flavus and Paecilomyces silvaticaA.5 percent in Pencillium notatum and 2.5 percent in Humicola grisea and Sepedonium chrysospermum. No detectable ED50 was observed in F. nivale .The concentration 10.5 percent was lethal for the eight tested fungi. Regarding the growth parameters, the spore germination, linear growth rate and dry biomass in all tested fungi decreased significantly with the increase in urea concentrations except in F .nivale where an increase in these growth criteria was recorded up to 4.5 percent urea. The pH value of the culture medium at the end of the incubation period shifted to alkaline side in A. fumigatus, A. niger and A .Flavus while no noticeable change was recorded in the media of the other fungi

Medium composition influencing single cell protein and cholesterol production by saccharomyces cerevisiae var carlsbergensis

Sugarcane molasses was found to be good carbon source for growth and production of single cell protein [SCP] by Saccharomyces cerevisiae var. carlsbergensis. Appreciable SCP yield was attained in the presence of molasses at 1 percent sugar concentration, whereas 5 percent was coupled with a good growth and minimal cholesterol content. The addition of 0.5 percent urea as a nitrogen source allowed better fermentation yield and suppressed cholesterol production. Further improvement of biomass and SCP production was achieved on adding 0.3 percent KH[2] PO[4] and 0.15 percent Mg SO[4]. 7H[2]O to the molasses medium. The comparative analysis of yeast cells grown on the synthetic [Phaff] medium with that grown on optimized molasses medium revealed a percentage increase in the later case of 415 percent SCP content, 527 percent peptides, 1018 percent amino acids, 201 percent ammonia, 394 percent total soluble nitrogen, 394 percent total nitrogen and 522 percent in dry weight gain. The cholesterol content decreased by 40 percent on molasses medium

Effect of benomyl on lignin degradation by some fungi

Fusarium oxysporum f. sp. vasinfectum, F.O. f.sp. Lycopersici and Aspergillus fumigatus could grow and metabolize cotton lignin when added as a sole carbon source to the culture media under control conditions [absence of fungicide]. The highest growth rate was achieved by A. fumigatus followed by F.O. f.sp. vasinfectum and F.O. f.sp. Lycopersici respectively. Conversely, the ligninolytic activity indicated that F.O. f.sp. Vasinfectum [cotton pathogen] solubilized larger quantities of phenols, as an intermediate product from degradation of cotton Iignin, followed by F.O. f.sp. lycopersici whereas A. fumigatus possessed the least ligninolytic activity. Addition of low levels of benomyl accelerated both growth and Iignin degradation by the three species, while the higher levels were inhibitory to them. The mycelial growth ceased at 1000, 800 and 200 ppm benomyl in A. fumigatus, F.O. f.sp. Vasinfectum and F.O. f.sp. Lycopersici, respectively, but Iignin degradation was detected at all fungicide concentrations. P-hydroxyberizoie, vanillic and syringic acids were spotted from the reaction mixture of the three fungi under control conditions. Vanillic acid was recorded at higher quantities and its metabolization by the three fungal species was continued at all benomyl concentrations. Addition of benomyl at low concentrations increased the solubilization of vanillic acid and significantly decreased that of p-hdyroxy benzoic and syringic acids. Higher benomyl concentrations inhibited vanillic acid production and totally arrested the formation of other phenolic acids