Methanol-Tolerant Lipase from Newly Isolated Saprochaete clavata 17B: Production, Characterization, and Application in Green Biodiesel Synthesis
DOI:
https://doi.org/10.48048/tis.2026.13231Keywords:
Biocatalyst, Biodiesel, Methanol-tolerant lipase, Saprochaete clavata, Statistical optimizationAbstract
The increasing demand for sustainable energy has driven interest in enzymatic biodiesel production, where lipases offer an eco-friendly alternative to chemical catalysts. This study aimed to optimize methanol-tolerant lipase production from yeast for biodiesel synthesis. Among four yeast strains tested, Saprochaete clavata 17B exhibited the highest extracellular lipase (ECL) activity at 457 U/L. Sequential optimization using the Taguchi method followed by response surface methodology–central composite design (RSM-CCD) was performed. The optimization experiments were conducted in two experiments with two different media formulations, using vegetable and alternative oils, under shaking conditions (200 rpm, 30 ± 2 °C, 120 h). The use of vegetable and alternative oils enhanced the ECL production to 5,277 and 3,260 U/L, respectively. Concentrated ECL obtained through ammonium sulfate saturation at 60-80% (w/v) showed a 7.4-fold increase in specific activity and was stable in the presence of 10% and 30% methanol. The lipase displayed optimal activity at pH 8.0 and room temperature, and its catalytic efficiency was evaluated in the transesterification of palm oil and Jatropha curcas seed oil, yielding 98% and 63% fatty acid methyl esters (FAME), respectively, while esterification of oleic acid produced 62% FAME. In conclusion, S. clavata 17B lipase represents a promising methanol-tolerant biocatalyst for industrial biodiesel applications, offering a sustainable alternative to chemical processes.
HIGHLIGHTS
- Lipase production by Saprochaete clavata 17B was significantly improved through Taguchi design and RSM-CCD optimization.
- Lipase activity increased by 7.4-fold after partial purification, with optimal performance at pH 8.0 and 30 ± 2 °C.
- Lipase remained stable after 3 h of incubation in the presence of 10% and 30% methanol, demonstrating methanol-tolerant properties.
- High biodiesel yields were obtained, reaching 98% from palm oil, 63% from Jatropha curcas seed oil, and 62% from oleic acid, demonstrating strong industrial potential.
GRAPHICAL ABSTRACT
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