Scale-up fermentation of Escherichia coli for the production of recombinant endoglucanase from Clostridium thermocellum

• Published in: Scientific reports Vol. 11; no. 1; p. 7145
• Main Authors: Shahzadi, Iram, Al-Ghamdi, Maryam A., Nadeem, Muhammad Shahid, Sajjad, Muhammad, Ali, Asif, Khan, Jalaluddin Azam, Kazmi, Imran
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 30-03-2021; Nature Publishing Group; Nature Portfolio
• Subjects: 631/326; 631/61; Cell Culture Techniques; Cellulase - biosynthesis; Cellulase - genetics; Cellulose; Clostridium thermocellum - genetics; E coli; Endoglucanase; Enzymes; Escherichia coli; Fermentation; Glycerol; Humanities and Social Sciences; Models, Statistical; multidisciplinary; pH effects; Recombinant Proteins - biosynthesis; Science; Science (multidisciplinary)
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-021-86000-z

Endoglucanase (EC 3.2.1.4) catalysing the hydrolysis of β-1.4-glycosidic linkage of cellulose molecules is an enzyme of tremendous industrial importance. The present study describes a response surface methodology based predicted model to deduce a set of fermentation conditions for optimum growth and activity of recombinant endoglucanase in
E. coli
BL21 (DE3). Numerous significant parameters including fermentation media composition, temperature (Celsius), pH and agitation rate (rpm) were analysed systemically by employing central composite design. This effort reports highly efficient recombinant endoglucanase overproduction (6.9 gl
−1
of biomass) with 30% expression by
E. coli
in modified M9NG media incubated at 37 °C and pH 7 agitated at 200 rpm. Addition of 3 mM glucose and 24 mM glycerol in the M9NG media has shown positive effect on the enzyme yield and activity. The CMCase activity experimentally estimated was found to be 1185 U/mg with the optimized parameters. The outcomes of both the responses by the predicted quadratic model were found in consensus with the obtained values. Our results well depicted the favourable conditions to further scale-up the volumetric yield of other relevant recombinant enzymes and proteins.