Jayachandran Dharanidaran, Chityala Sushma, Prabhu Ashish A, Dasu Veeranki Venkata
Biochemical Engineering Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology (IIT) Guwahati, Guwahati, 781039, Assam, India.
Biochemical Engineering Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology (IIT) Guwahati, Guwahati, 781039, Assam, India.
Protein Expr Purif. 2019 May;157:1-8. doi: 10.1016/j.pep.2019.01.002. Epub 2019 Jan 4.
Reverse micellar extraction (RME), a liquid-liquid based separation is a versatile tool for protein purification. A statistical approach was employed for the purification of recombinant glutaminase free anti-cancerous enzyme viz., l-asparaginase II to evaluate the effects of RME in current study. The cationic system (CTAB/iso-octane/hexanol/butanol) was used in RME to optimize both forward and backward protein extraction efficiency. By adapting Taguchi's orthogonal array (OA), maximum forward extraction efficiency (FEE) of 86.98% with 84.82% enzyme activity recovery and 1.04 times purification fold achieved with the optimized parameters. Under the optimal levels, the back extraction efficiency (BEE) was observed to be 96.97% with 93.07% enzyme activity recovery and 1.38 times purification fold. Further, mass transfer kinetic studies of RME indicated the mass transfer coefficients of forward and backward extraction to be 0.049 min and 0.036 min respectively.
反胶束萃取（RME）是一种基于液-液的分离方法，是蛋白质纯化的一种通用工具。本研究采用统计方法纯化重组无谷氨酰胺酶的抗癌酶，即L-天冬酰胺酶II，以评估RME的效果。在RME中使用阳离子体系（CTAB/异辛烷/己醇/丁醇）来优化正向和反向蛋白质萃取效率。通过采用田口正交阵列（OA），在优化参数下实现了86.98%的最大正向萃取效率（FEE），酶活性回收率为84.82%，纯化倍数为1.04倍。在最佳水平下，反向萃取效率（BEE）为96.97%，酶活性回收率为93.07%，纯化倍数为1.38倍。此外，RME的传质动力学研究表明，正向和反向萃取的传质系数分别为0.049 min和0.036 min。
