Biodiesel production was investigated by hydroesterification using enzymatic catalysis in both hydrolysis and esterification reactions. Soybean oil resulting from food frying processes (WCO) was used as feedstock. Lipase from Geotrichum candidum was produced by submerged fermentation and used without prior purification as biocatalyst in WCO hydrolysis, which was optimized using a factorial design. A complete hydrolysis of WCO (44.1% mass fraction) was obtained after 80 min of reaction at 40 °C, performed in an emulsifier-free system with no buffer (40 °C and 900 rpm). In a subsequent step, FFA obtained from the hydrolysis reaction were esterified (40 °C, 200 rpm) using ethanol as acyl-acceptor (1:1.5) and Pseudomonas fluorescens lipase (PFL) immobilized on styrene-divinylbenzene (SDB) as biocatalyst (15% m/v). This biocatalyst was recycled for 6 cycles with no significant activity losses. The biodiesel composition was found by gas chromatography, and proton nuclear magnetic resonance (¹H NMR) spectra and Fourier transform infrared spectroscopy (FTIR) were used to verify the quality and purity of biodiesel. Enzymatic biodiesel achieved FAEE content of 94.1 ± 0.5% and no traces of monoacyglycerol (MAG), diacylglycerol (DAG), triacylglycerol (TAG) or free glycerol were observed, indicating that the hydroesterification route produced high-quality biodiesel, despite the fact that WCO was used as feedstock.

通过在水解和酯化反应中使用酶催化进行的加氢酯化研究了生物柴油的生产。食品油炸过程（WCO）产生的豆油用作原料。来自白色假丝酵母的脂肪酶是通过深层发酵生产的，无需事先纯化即可用作WCO水解中的生物催化剂，并通过因子设计对其进行了优化。在40°C反应80分钟后，在无缓冲剂的无乳化剂系统（40°C和900 rpm）中进行WCO的完全水解（质量分数为44.1％）。在随后的步骤中，使用乙醇作为酰基受体（1：1.5）和荧光假单胞菌将水解反应所得的FFA酯化（40°C，200 rpm）脂肪酶（PFL）作为生物催化剂固定在苯乙烯-二乙烯基苯（SDB）上（15％m / v）。将该生物催化剂再循环6个循环，没有明显的活性损失。通过气相色谱法发现了生物柴油的组成，并利用质子核磁共振波谱（¹ H NMR）和傅里叶变换红外光谱（FTIR）来验证生物柴油的质量和纯度。酶促生物柴油的FAEE含量为94.1±0.5％，没有观察到痕量的单酰甘油（MAG），二酰甘油（DAG），三酰甘油（TAG）或游离甘油，这表明加氢酯化途径可生产出高质量的生物柴油，尽管事实是WCO用作原料。