Abstract: Alkane-based biodiesel is considered the next generation of biodiesel owing to its potential environmental benefits and the fact that it exhibits much higher specific caloric values than traditional biodiesel. However, the formidable obstacle impeding the commercialization of this cutting-edge fuel alternative lies in the cost associated with its production. In this study, an engineered strain Escherichia coli (E. coli) showcasing harmonized coexpression of a lipase (from Thermomyces lanuginosus lipase, TLL) and a fatty acid photodecarboxylase (from Chlorella variabilis, CvFAP) was first constructed to transform triglycerides into alkanes. The potential of E. coli BL21 (DE3)/pRSFDuet-1-TLL-CvFAP for alkane synthesis was evaluated with tripalmitin as a model substrate under various process conditions. Following a comprehensive examination of the reaction parameters, the scope of the biotransformation was expanded to ‘real’ substrates (vegetable oils). The results showed that bioderived oils can be transformed into alkanes with high yields (0.80-10.20 mmol·L^-1) under mild conditions (35 ℃, pH 8.0, and 36 h) and blue light illumination. The selected processes were performed on an increased lab scale (up to 100 ml) with up to 24.77 mmol·L^-1 tripalmitin, leading to a yield of 18.89 mmol·L^-1 pentadecane. With the employment of a method for efficiently producing alkanes under mild conditions and a simple procedure to isolate alkanes from the reaction system, the utilization of sustainable biomass as a fundamental feedstock emerges as the primary solution to lower the cost of alkane-based biodiesel. Thus, this study proposes a readily implementable and highly effective approach for alkane-based biodiesel production.

Key words: Alkane-based biodiesel, Renewable biomass, Co-overexpression, Lipase, Photodecarboxylase

摘要： Alkane-based biodiesel is considered the next generation of biodiesel owing to its potential environmental benefits and the fact that it exhibits much higher specific caloric values than traditional biodiesel. However, the formidable obstacle impeding the commercialization of this cutting-edge fuel alternative lies in the cost associated with its production. In this study, an engineered strain Escherichia coli (E. coli) showcasing harmonized coexpression of a lipase (from Thermomyces lanuginosus lipase, TLL) and a fatty acid photodecarboxylase (from Chlorella variabilis, CvFAP) was first constructed to transform triglycerides into alkanes. The potential of E. coli BL21 (DE3)/pRSFDuet-1-TLL-CvFAP for alkane synthesis was evaluated with tripalmitin as a model substrate under various process conditions. Following a comprehensive examination of the reaction parameters, the scope of the biotransformation was expanded to ‘real’ substrates (vegetable oils). The results showed that bioderived oils can be transformed into alkanes with high yields (0.80-10.20 mmol·L^-1) under mild conditions (35 ℃, pH 8.0, and 36 h) and blue light illumination. The selected processes were performed on an increased lab scale (up to 100 ml) with up to 24.77 mmol·L^-1 tripalmitin, leading to a yield of 18.89 mmol·L^-1 pentadecane. With the employment of a method for efficiently producing alkanes under mild conditions and a simple procedure to isolate alkanes from the reaction system, the utilization of sustainable biomass as a fundamental feedstock emerges as the primary solution to lower the cost of alkane-based biodiesel. Thus, this study proposes a readily implementable and highly effective approach for alkane-based biodiesel production.

关键词: Alkane-based biodiesel, Renewable biomass, Co-overexpression, Lipase, Photodecarboxylase