Optimization of Conditions for Maximal Production of Recombinant Thermostable Cellulase from Thermotoga naphthophila using E. coli BL21-CodonPlus (DE3) as Expression Host
Optimization of Conditions for Maximal Production of Recombinant Thermostable Cellulase from Thermotoga naphthophila using E. coli BL21-CodonPlus (DE3) as Expression Host
Aisha Khalid1, Muhammad Tayyab1,*, Abu Saeed Hashmi1, Tahir Yaqub2, Ali Raza Awan1, Muhammad Wasim1, Shagufta Saeed1, Sehrish Firyal1 and Abdul Rauf Shakoori3
ABSTRACT
Current study was designed for the development of an economic and environment friendly mechanism for the production of thermostable cellulase. Production of cellulase was focused due to its diverse range of application in industry. In the present study, conditions were optimized for the maximal production of recombinant thermostable cellulase from Thermotoga naphthophila using BL21-CodonPlus (DE3) cells as expression host and pET28a as expression vector. Effect of various concentration of Isopropyl β-D-1-thiogalactopyranoside (IPTG), post induction time, effect of temperature and pH were examined for the maximal production of recombinant cellulase. The effect of supplementation of LB medium with additional carbon and nitrogen sources was also analyzed for maximal production of recombinant protein. Higher level enzyme activity was recorded at 25°C, pH 7.0 when the cells were induced with 0.5 mM IPTG with 22h post induction incubation. Supplementation of LB medium with 1% glucose and yeast extract enhanced the production of recombinant thermostable cellulase. Enzyme showed strong potential for its use in paper and poultry feed industry. Under the optimal conditions we could able to produce 48 U/mL of recombinant cellulase.
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August 2019
Vol. 51, Iss. 4, Pages 1203-1598
