Abstract

This study reports cellulase production by Bacillus paralicheniformis using Plumeria alba leaf powder under submerged fermentation with a focus on systematic bioprocess optimization. Physical parameters were first optimized using a one-factor-at-a-time (OFAT) approach, followed by optimization of yeast extract, MgSO4 and (NH4)2SO4 via a central composite design (CCD) and response surface methodology (RSM). An artificial neural network (ANN) with a 5:3:1 network trained by the Levenberg–Marquardt algorithm further improved prediction of carboxylmethylcellulase (CMCase) and filter paper cellulase (FPase) activities. This study is the first to exploit Plumeria alba leaf powder as an untapped, low-cost lignocellulosic substrate for cellulase production by B. paralicheniformis and uniquely benchmarks RSM against ANN-based modeling to identify superior predictive frameworks for bioprocess optimization. Under optimized conditions (24 h, 4% w/v substrate, 1% v/v inoculum), the maximum FPase and CMCase activities reached 60.53 IU/mL/min and 332.10 IU/mL/min respectively. Partial characterization showed optimum FPase and CMCase activities at 50 °C and 70 °C, respectively, at pH 7.5. Enzymes also showed activation by NaCl and some select solvents while tolerating a broad range of metal ions. The enzymatic hydrolysis of P. alba biomass released 59.42 mg/mL total reducing sugars after 8hr, confirming efficient saccharification from a low-cost feedstock. The ANN model (R2 = 97.59% for CMCase; 85.95% for FPase) outperformed RSM (R2 = 85.95% and 78.25%, respectively), while radial basis function optimization reached 99.99%. These findings highlight B. paralicheniforms cellulase as a promising biocatalyst for biorefinery applications and demonstrate the value of integrating RSM and ANN for process optimization.

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