Kanmani P, Karthik S, Aravind J, Kumaresan K
Department of Biotechnology, Kumaraguru College of Technology, Coimbatore 641049, India.
ISRN Biotechnol. 2012 Oct 22;2013:528708. doi: 10.5402/2013/528708. eCollection 2013.
The optimization of extracellular lipase production by Fusarium isolani strain SKWF7 isolated from dairy wastewater was carried out in this study. Initially, the physicochemical factors significantly influencing enzyme production were studied by varying one-factor-at-a-time (OFAT). A mesophilic temperature of 40°C, alkaline pH of 8, and incubation period of 72 hours were found to be the optimal conditions for lipase production. Among the media components, the disaccharide sucrose acted as the best carbon source; palm oil as the best inducing lipid substrate; casein and (NH4)2SO4 as the best organic and inorganic nitrogen sources; Ca(2+) ion as the best trace element. In the next phase of work, statistical optimization of medium components was performed by employing the Box-Behnken design of Response Surface Methodology (RSM). The optimum concentrations of three significant factors, namely, palm oil, (NH4)2SO4, and CaCO3 were determined by this method to be 5% (v/v), 5.5 g/L, and 0.1 g/L, respectively. RSM-guided design of experiments resulted in a maximum lipase production of 73.3 U/ml, which is a 1.7-fold increase in comparison with that obtained in the unoptimized medium. These results point towards the success of the model in developing a process for the production of lipase, an enzyme of enormous industrial significance.
本研究对从乳品废水中分离出的茄病镰刀菌菌株SKWF7胞外脂肪酶的生产进行了优化。最初,通过单因素变化法(OFAT)研究了对酶生产有显著影响的理化因素。发现40°C的嗜温温度、pH值为8的碱性条件以及72小时的培养时间是脂肪酶生产的最佳条件。在培养基成分中,二糖蔗糖是最佳碳源;棕榈油是最佳诱导脂质底物;酪蛋白和硫酸铵是最佳有机和无机氮源;钙离子是最佳微量元素。在工作的下一阶段,采用响应面法(RSM)的Box-Behnken设计对培养基成分进行了统计优化。通过该方法确定的三个显著因素,即棕榈油、硫酸铵和碳酸钙的最佳浓度分别为5%(v/v)、5.5 g/L和0.1 g/L。RSM指导的实验设计使脂肪酶的最大产量达到73.3 U/ml,与未优化培养基相比提高了1.7倍。这些结果表明该模型在开发脂肪酶生产工艺方面取得了成功,脂肪酶是一种具有巨大工业意义的酶。