Geoffry Kiptoo, Achur Rajeshwara N
Department of Biochemistry, Kuvempu University, Shankaraghatta, 577451 Shimoga, Karnataka, India.
J Genet Eng Biotechnol. 2018 Dec;16(2):327-334. doi: 10.1016/j.jgeb.2018.04.003. Epub 2018 Apr 27.
Among different sources of lipases, fungal lipases have continued to attract a wide range of applications. Further, halophilic lipases are highly desirable for biodiesel production due to the need to mitigate environmental pollution caused as result of extensive use of fossil fuels. However, currently, the high production cost limits the industrial application of lipases. In order to address this issue, we have attempted to optimize lipase production by NFCCL 4084 and using palm oil mill effluent (POME) based medium. The production was optimized using a combinatory approach of Plackett-Burman (PB) design, one factor at a time (OFAT) design and face centred central composite design (FCCCD). The variables (malt extract, (NH)SO, CaCl, MgSO, olive oil, peptone, KHPO, NaNO, Tween-80, POME and pH) were analyzed using PB design and the variables with positive contrast coefficient were found to be KHPO, NaNO, Tween-80, POME and pH. The significant variables selected were further analyzed for possible optimum range by using OFAT approach and the findings revealed that KHPO, NaNO, and Tween-80 as the most significant medium components, and thus were further optimized by using FCCCD. The optimum medium yielded a lipase with an activity of 7.8 U/ml, a significant 3.2-fold increase compared to un-optimized medium. The present findings revealed that POME is an alternative and suitable substrate for halophilic lipase production at low cost. Also, it is clearly evident that the combinatory approach employed here proved to be very effective in producing high activity halophilic lipases, in general.
在不同来源的脂肪酶中，真菌脂肪酶一直吸引着广泛的应用。此外，由于需要减轻因大量使用化石燃料而造成的环境污染，嗜盐脂肪酶在生物柴油生产中非常受欢迎。然而，目前高生产成本限制了脂肪酶的工业应用。为了解决这个问题，我们尝试通过NFCCL 4084优化脂肪酶的生产，并使用基于棕榈油厂废水（POME）的培养基。使用Plackett-Burman（PB）设计、一次一个因素（OFAT）设计和中心复合设计（FCCCD）的组合方法对生产进行了优化。使用PB设计分析了变量（麦芽提取物、(NH)SO、CaCl、MgSO、橄榄油、蛋白胨、KHPO、NaNO、吐温-80、POME和pH），发现具有正对比系数的变量是KHPO、NaNO、吐温-80、POME和pH。通过OFAT方法进一步分析所选的显著变量的可能最佳范围，结果表明KHPO、NaNO和吐温-80是最显著的培养基成分，因此通过FCCCD进一步优化。最佳培养基产生的脂肪酶活性为7.8 U/ml，与未优化的培养基相比显著提高了3.2倍。目前的研究结果表明，POME是一种低成本生产嗜盐脂肪酶的替代且合适的底物。此外，很明显，这里采用的组合方法总体上被证明在生产高活性嗜盐脂肪酶方面非常有效。
