The Use of Response Surface Methodology as a Statistical Tool for Media Optimization in Lipase Production from the Dairy Effluent Isolate Fusarium solani.

ISRN biotechnology Pub Date : 2012-10-22 eCollection Date: 2013-01-01 DOI:10.5402/2013/528708
P Kanmani, S Karthik, J Aravind, K Kumaresan
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Abstract

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.

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将响应面法作为一种统计工具,用于优化乳制品污水菌株 Fusarium solani 生产脂肪酶的培养基。
本研究对分离自乳品废水的分离镰刀菌株 SKWF7 的胞外脂肪酶生产进行了优化。首先,通过一次性改变一个因素(OFAT)来研究对产酶有显著影响的理化因素。结果发现,40°C 的中嗜酸性温度、8°C 的碱性 pH 值和 72 小时的培养期是产生脂肪酶的最佳条件。在培养基成分中,蔗糖双糖是最佳碳源;棕榈油是最佳诱导脂质底物;酪蛋白和(NH4)2SO4 是最佳有机和无机氮源;Ca(2+)离子是最佳微量元素。在下一阶段的工作中,采用响应面法(RSM)的箱-贝肯设计对培养基成分进行了统计优化。通过这种方法确定了棕榈油、(NH4)2SO4 和 CaCO3 这三个重要因素的最佳浓度分别为 5% (v/v)、5.5 g/L 和 0.1 g/L。通过 RSM 指导下的实验设计,脂肪酶的最大产量为 73.3 U/ml ,比未优化培养基的产量提高了 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,而脂肪酶是一种具有重大工业意义的酶。
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