An α-1,4-glycosidic bonds galactoses pectin, mainly composed of a D-galacturonic acid chain, are important biomaterial widely used in industries. Utilizing this material, a bioprocess, including the biocatalysis pectinase, is often needed. Pectinase production was optimized in 7 days SSF at 37°C, and the pectinase activities were daily measured by the method of Somogy-Nelson. The optimum pectinase production was 0.166 U/ml on the fourth day SSF. Purification using open column ion exchange chromatography DEAE cellulose DE-52 resulted in 1030.9 folds of pectinase purity with a yield of 25.9%. The enzyme was at optimal activity at pH six and attended stable in the pH range of 5.5-8, while optimal activity at a temperature of 50°C and was stable in the range of 30-45°C. The pectinase activity increased by 120% with the addition of 10 mM Mg2+, and 95% retained when 10 mM Ca2+ was added. The presence of 10 mM Na+, K+, and Fe2+ resulted in a slight effect of activity at 85%, 83%, and 78%. However, it was strongly inhibited by 10 mM Al3+ and retained 25%. Based on the results above, the microbial utilization of coffee pulp waste by ISH16 bacteria pectinolytic is one opportunity to produce valuable pectinase with low-cost production, so comprehensive examination in large-scale production is needed too. In this paper, all research detail steps were described.
Cellulase has been widely used in many applications of industries and is commonly produced by a cellulase-producing-fungus using production substrates from agricultural wastes biomass. In the wet processing of the coffee, up to 63% of coffee pulp rich-cellulose has been released. This potential waste allows being used as a substrate for enzyme production. This study aims to produce cellulase using coffee pulp by Solid-State Fermentation from isolates of Aspergillus sp. VT12. A solid-state fermentation medium of coffee pulp was inoculated with Aspergillus sp. VT12 and incubated at 37 °C for 5 days, the culture produce a maximum cellulase activity of 0.52 U/ml against 0.5% carboxymethyl cellulose substrate. For the purification step, the crude cellulase obtained was dialyzed on a cellulose tube membrane 12-14 kDa and then loaded into open column anion exchanger DEAE Cellulose DE-52 chromatography with NaCl gradient 0-0.6M used. In this step, the activity of purified cellulase obtained was 1.00 U/ml with a yield of 51.86%. For all steps of dialysis and cellulase measurement activity, including purification steps, the 20 mM acetate buffer pH 5 was used. This research revealed that the potential coffee pulp waste can be used as a substrate for cellulase production. However, in an economical view, production efficiency still needs to be optimized and the purity of the enzyme as well.
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