OPTIMIZATION OF CARBOXY METHYL CELLULASE PRODUCTION BY CLADOSPORIUM SP. WITH FOOD WASTE IN BAGASSE SOLID STATE FERMENTATION USING RESPONSE SURFACE METHODOLOGY

Abstract

Enzyme production costs should be reduced by using inexpensively available substrates. In this, we determine the effects of various medium components and food processing waste on Cladosporium sp. production of CMCase in SSF. Five factors were tested with a three-level Box-Wilson factorial, central composite design (CCD) to determine how they impacted Cladosporium sp. NCIM 901's ability to produce CMCase. For the model, the R² (coefficient of determination) value was 0.9349. Based on the process findings, the optimal medium for higher enzyme synthesis had wheat bran (4 g/L), cacl₂ (2 g/L), urea (7.5 g/L) and rice bran (8.76 g/L), with the medium's original pH at 5.5. The model's accuracy is confirmed by the experimental result of CMCase, which is 23.42 U/gds and equivalent to the expected yield of 22.31 U/gds. The accuracy of the current experimental design has been verified by experimental validation. The enzyme's properties indicated that its greatest activity occurred at pH 5.5 and that its ideal temperature was about 50 °C. The enzyme is more active toward CMC and less active toward filter paper. CaCl₂ and MgCl₂ increased the enzyme's activity. Enzymatic hydrolysis of waste-derived polysaccharides into simple sugars is necessary for the industrial manufacture of bioethanol. The efficacy of the produced Cladosporium sp. cellulase (CMCase) enzyme to saccharify pearl millet grass results shows that H₂SO₄ treated substrate gave more fermentable sugars compared to the untreated substrate. Results suggest that the indigenous produced-CMCase can be efficiently used for lignocellulosic biomass saccharification (55.40%) compared to the untreated sample (37.10%) in 48 h, which will eventually reduce the cost utilized for purchasing commercial enzymes.