POTENTIAL CRUDE ENZYME CELLULOSE Bacillus sp. FROM TEMPEH GEMBUS FERMENTATION FOR PRODUCTION OF MICRO-MEMBRANE CELLULOSE/CHITOSAN

• Main Author: Aji Saputra, Tiar
• Format: Theses
• Language: Indonesia
• Subjects: Ilmu hayati ; Biologi
• Online Access: https://digilib.itb.ac.id/gdl/view/35227
• Institution: Institut Teknologi Bandung

Soybean residue is a byproduct from tofu industry. Soybean residue has the potential nutrients such as fiber and starch. This potential can be used as a substrate for fungi to produce cellulose and chitin. Microfungi and soybean fibers can be used to produe Cryctaline Micro Cellulose (CMC), Fiber Micro Cellulose (FMC) and other cellulose polymers. Previous research through the addition of inoculum Rhizopus microsporus var. oligosporus in the production of tempeh can increase the fiber content of soybean residue in Tempeh. With that potential, this research analyzes and optimizes the production of micro-cellulose membranes using crude extracts of cellulose enzymes produced by indigenous of bacteria on production process. Micro-cellulose particles formed are then made into membranes by gelatinization using cross-linking agent chitosan. Production of cellulase enzyme was achieved using submerged fermentation system with Carboxymethyl Cellulose (CMC) substrate. Protein concentration was measured and crude cellulase enzyme was characterized and its activity was tested. Crude enzyme with various level of enzyme activity was used to produce microcellulose. In this research, randomized block design (RBD) was applied with three factors such as enzyme activity level (2.5 U/g, 5 U/g, and 12.5 U/g substrate), incubation time (48 hours and 96 hours) and substrate pre-treatment using H2O2 (2 hours and 12 hours, at 80oC). The experiment was carried out at 45oC and agitation of 150 rpm, with citrate buffer pH 4.8. Cellulose particles produced were measured by Particle Size Analyzer (PSA) with H2O solvent and visualized by Franhauer model. Cellulose particles were then tested for transmitant and absorb group function using FTIR (Fourier-transform Infrared Spectroscopy). In the final stage, cellulose membrane gelatinase was carried out by adding chitosan. The membrane formed is characterized by absorption, water content (KA) and visual observation using SEM (Scanning Electron Microscopy). Cellulase enzyme produced by Bacillus sp. harvested at 60 hours of production with total protein concentration of 0.16-0.19 mg/ml and enzyme activity 0.26-0.28 U/mL and 0.096-0.10 FPU/mL. Enzyme has a specific activity at 45oC and pH 4.8. The enzyme activity produced is equivalent to 2.4-2.5 U/ g using CMCase method. The treatment results in the cellulose particle production stage showed significant difference (with a 95% confidence level) between three treatment factors with a significance value <0.05. The best particle size produced was 0.678 ?m at activity level of 12.5 U/g, 96 hours of incubation, and 12 hours of H2O2 pre-treatment. However, the treatment of incubation time compared to the level of enzyme activity was not significantly different from the significance value of 0.384. Particle testing using FTIR shows increase of transmission of hydrogen bonds (O-H) in the range 3427.51 to 3468.01 and transmission of glycosidic bonds with a spectrum of 1035.77 to 1066.06. SEM visualization of the membrane show pore formation at the addition of 10% cellulose and produced a membrane with the lowest water content of 27.43%. Based on the results in this research, the crude cellulase produced by Bacillus sp. can be used for the production of micro-cellulose using treatment with 12.5 U / g of enzyme activity, incubation time of 96 hours, and H2O2 pre-treatment for 12 hours. Membrane production can be produced with the composition such as 1%(w/v) chitosan, 10%(v/v) glycerol, 10%(w/v) microcellulose particles and dH2O solvent as a pre-rehydration membrane formulation.