由於近年來原油價格的飆漲，儲存量的持續遞減和溫室效應的日益嚴重，利用生質燃料來替代石化燃料已成為不可避免的趨勢。因此，新能源的開發無不受到世界各國的重視。生化柴油是一種可再生的生物資源，因其對環境的污染性低、生物可降解性以及柴油引擎可無須任何修改即可直接使用而受到世人的矚目。目前，生化柴油的生產方法主要可分為化學製程（鹼、酸觸媒）和酵素製程。
本研究利用酵素水解－酸酯化複合製程來生產生化柴油。第一階段，Candida rugosa脂肪分解酶利用二元固定化法固定在幾丁聚醣擔體上應用於黃豆油水解。第二階段則是將酵素水解所得的脂肪酸進行酸觸媒酯化反應生產生化柴油。二元固定化酵素在40℃、pH8以及油脂/水/固定化酵素組成為10/5/3（w/w/w）的環境下，水解五小時，轉化率可達90 %。酸酯化反應則在50℃，2.5%硫酸和甲醇/脂肪酸的莫耳比為15/1的較適化條件下，反應半小時和12小時，生化柴油轉化率分別為83 %和99%。由油脂的性質分析實驗來看，利用黃豆油做為原料，經由酵素水解－酸酯化複合製程所得到的生化柴油，在所測試的性質中皆可符合美國ASTM所訂立的規範(ASTM D6751-02)。由實驗結果可知，酵素水解－酸酯化複合製程是一個適合生產生化柴油的方法。

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