In the present study, Proteus vulgaris lipase (PVL) was engineered using directed evolution to increase methanol tolerance so that it would be more tolerant and efficient for harsh conditions employed in biodiesel synthesis, which is limiting their industrial use. The influence of ultrasound under different experimental conditions on the biodiesel conversion yield using methanolysis of non-edible neem oil was also emphasized. A special attention was also paid to the immobilization of lipase on Polysulfone (PS) beads and comparative studies with industrially used Burkholderia cepacia lipase. The Engineered Proteus vulgaris lipase showed >80% activity after 3 h when incubated in 50% methanol with simultaneous sonication. The lipase retained improved longevity (~70% residual activity) over wild-type PVL over repeated use.

在本研究中，普通变形杆菌Proteus vulgaris脂肪酶（PVL）是通过定向进化设计的，以提高甲醇耐受性，因此对于生物柴油合成中使用的苛刻条件将具有更大的耐受性和效率，这限制了其工业用途。还强调了超声波在不同实验条件下对不可食用印em油甲醇分解的生物柴油转化率的影响。还特别注意了脂肪酶在聚砜（PS）磁珠上的固定以及与工业上使用的伯克霍尔德酒原洋葱脂肪酶的比较研究。改造的普通变形杆菌当在50％甲醇中同时超声处理时，脂肪酶在3小时后显示> 80％的活性。与野生型PVL相比，脂肪酶在重复使用后具有更长的使用寿命（约70％的残留活性）。