Promoting Magnusiomyces spicifer AW2 Cell-Bound Lipase Production by Co-culturing with Staphylococcus hominis AUP19 and Its Application in Solvent-Free Biodiesel Synthesis.

Yeast-bacterium interaction has recently been investigated to benefit the production of cell-bound lipases (CBLs). Staphylococcus hominis AUP19 supported the growth of Magnusiomyces spicifer AW2 in a palm oil mill effluent (POME) medium to produce CBLs through a bioremediation approach, including oil and grease (O&G) and chemical oxygen demand (COD) removals. This research used the yeast-bacterium co-culture to optimize CBLs and cell biomass (CBM) productions through bioremediation using the statistical Plackett-Burman design and response surface methodology-central composite design. The CBLs were finally applied in biodiesel synthesis. The CBM of 13.8 g/L with CBLs activity at 3391 U/L was achieved after incubation at room temperature (RT, 30 ± 2 °C) for 140 h in 50% POME medium, pH 7.0, containing 1.23% (w/v) ammonium sulfate. Bacterium promoted yeast growth to achieve bioremediation with 87.9% O&G removal and 84.5% COD removal. Time course study showed that the CBLs activity was highest at 24 h cultivation (4103 U/L) and retained 80% and 60% of activities at 4 °C and RT after 5 weeks of storage. The CBLs application successfully yielded 77.3% biodiesel from oleic acid (esterification) and 86.4% biodiesel from palm oil (transesterification) within 72 h in solvent-free systems. This study highlights that yeast-bacterium co-culture and POME should receive more attention for potential low-cost CBLs production through bioremediation, i.e., O&G and COD removals, while the CBLs as biocatalysts are promising for significant contribution to an effective strategy for economic green biodiesel production.

The Occurrence of Triple Catalytic Characteristics of Yeast Lipases and Their Application Prospects in Biodiesel Production from Non-Edible Jatropha curcas Oil in a Solvent-Free System.

Extracellular and cell-bound lipase-producing yeasts were isolated from the palm oil mill wastes and investigated for their potential uses as biocatalysts in biodiesel production. Twenty-six yeast strains were qualitatively screened as lipase producers. From those yeast strains, only six were selected and screened further for quantitative lipase production.The phylogenetic affiliations of the yeast strains were confirmed by investigating the D1/D2 domains of 26S rDNA and ITS1-5.8S-ITS2 molecular regions of the six yeast strains selected as potent lipase producers. The three yeast strains A4C, 18B, and 10F showed a close association with Magnusiomyces capitatus. Two yeast strains (17B and AgB) had a close relationship with Saprochaete clavata, whereas the strain AW2 was identified as Magnusiomyces spicifer. Three main catalytic activities of the yeast lipases were evaluated and Magnusiomyces capitatus A4C, among the selected lipase-producing yeasts, had the highest extracellular lipolytic enzyme activity (969 U/L) with the cell-bound lipolytic enzyme activity of 11.3 U/gdm. The maximum cell-bound lipolytic activity (12.4 U/gdm) was observed in the cell-bound lipase fraction produced by Magnusiomyces spicifer AW2 with an extracellular lipolytic enzyme activity of 886 U/L. Based on the specific hydrolytic enzymatic activities, the cell-bound lipases (CBLs) from the three yeast strains M. capitatus A4C, M. spicifer AW2, and Saprochaete clavata 17B were further investigated for biodiesel production. Among them, the CBL from M. spicifer AW2 synthesized the most FAME (fatty acid methyl esters) at 81.2% within 12 h indicating that it has potential for application in enzymatic biodiesel production.