Patel Radhika, Prajapati Vimal, Trivedi Ujjval, Patel Kamlesh
P. G Department of Biosciences, Sardar Patel University, Sardar Patel Maidan, Vadtal Road, Satellite Campus, Bakrol, Gujarat 388315 India.
Aspee Shakilam Biotechnology Institute, Navsari Agricultural University, Athwa Farm, Ghod Dod Road, Surat, Gujarat 395007 India.
3 Biotech. 2020 Dec;10(12):508. doi: 10.1007/s13205-020-02501-0. Epub 2020 Nov 5.
Organic solvent-tolerant lipase-producing microorganisms were isolated from petrol spilled soil. From ten morphologically distinct lipase-producing bacterial isolates, highest amount of lipase-producing isolate UBT1 was identified as sp. using 16S rRNA gene sequencing (NCBI Accession No: MH879815). An increase in lipase production from 42 U/mL to 243 U/mL was obtained when different deoiled seed cakes were supplemented instead of olive oil in the medium. Further optimization of media components by the statistical approach assisted in discerning the main influencing media components and their optimum concentrations. Nine components glucose, castor seedcake, potassium nitrate, gum arabic, calcium chloride, magnesium sulphate, potassium di-hydrogen phosphate, dipotassium hydrogen phosphate, and ferric chloride were selected for Plackett-Burman design. The optimum concentrations of three significant selected components for the lipase production were found to be 0.025 gm% glucose, 0.002 gm% calcium chloride, and 0.2 gm% potassium di-hydrogen phosphate as determined by Response Surface Methodology. Increase in lipase production with 292.29 U/mL was achieved in the media containing optimized components and 2 gm% deoiled castor seed cake. Purification studies with ammonium sulphate precipitation, dialysis, and gel permeation chromatography resulted in 77.54% recovery with 5.77-fold partially purified lipase. The residual activity of lipase in 50 and 75% concentration of n-hexane among other solvents after 24 h was 105.05 and 90.42%, respectively, indicating its solvent tolerance. The present study reports the isolation of organic solvent-tolerant lipase-producing sp. UBT1, optimization of the culture media for lipase production using the deoiled castor seed cake, and its partial purification.
从被石油污染的土壤中分离出了耐有机溶剂的产脂肪酶微生物。在十个形态各异的产脂肪酶细菌分离株中，通过16S rRNA基因测序（NCBI登录号：MH879815），产脂肪酶量最高的分离株UBT1被鉴定为 属。当在培养基中添加不同的脱油籽饼代替橄榄油时，脂肪酶产量从42 U/mL增加到了243 U/mL。通过统计方法对培养基成分进行进一步优化，有助于识别主要影响培养基的成分及其最佳浓度。选择了葡萄糖、蓖麻籽饼、硝酸钾、阿拉伯胶、氯化钙、硫酸镁、磷酸二氢钾、磷酸氢二钾和氯化铁九种成分进行Plackett-Burman设计。通过响应面法确定，所选三种显著促进脂肪酶产生的成分的最佳浓度分别为0.025 gm%葡萄糖、0.002 gm%氯化钙和0.2 gm%磷酸二氢钾。在含有优化成分和2 gm%脱油蓖麻籽饼的培养基中，脂肪酶产量增加到了292.29 U/mL。通过硫酸铵沉淀、透析和凝胶渗透色谱法进行纯化研究，得到了回收率为77.54%、部分纯化了5.77倍的脂肪酶。在24小时后，脂肪酶在50%和75%浓度的正己烷以及其他溶剂中的残留活性分别为105.05%和90.42%，表明其具有耐溶剂性。本研究报道了耐有机溶剂的产脂肪酶 属菌株UBT1的分离、使用脱油蓖麻籽饼优化脂肪酶生产培养基及其部分纯化。
