Enhanced production of novel halostable recombinant endoglucanase derived from the metagenomic library using fed-batch fermentation. (March 2019)

• Record Type: Journal Article
• Authors: Kumar, Narender; Sudan, Sarabjeet Kour; Garg, Roma; Sahni, Girish
• Is Part Of: Process biochemistry. Volume 78(2019)
• Page Start: 1
• Page End: 7
• Publication Date: 2019-03
• Subjects: DOC dissolved oxygen concentration -- DNS 3, 5-dinitrosalicylic acid -- CMC carboxymethylcellulose; Endoglucanase -- Halostable -- Metagenomic -- Fed-batch fermentation; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2018.12.033
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: Novel endoglucanase was isolated from soil metagenomic library. 40 fold increase in yield was obtained using fed- batch strategies as compared to shake flask. Purified endogluconase was halostable and halotolerant as it retained 100% activity after 30 days incubation in presence of salts. The purified enzyme was also found to be stable in presence of ions, organic solvents, surfactant and detergents. Abstract: A novel halostable endoglucanase Cel5R obtained from soil metagenomic library was cloned and expressed in Escherichia coli BL21 (DE3). In this study, we investigated the effect of dissolved oxygen concentration (DOC) on plasmid stability and effect of complex nitrogen source on the production of protein. With the increase in DOC from 30% to 50%, the production of protein also increased from 500 mg/l to 3 g/l. Besides, the addition of yeast extract and tryptone increased the production of endoglucanase up to 5 g/l. Overall, the production of endoglucanase showed an increase of 40-folds from shake flask to fed-batch fermentation. Biochemical characterization of the purified enzyme revealed that the enzyme retained its 100% catalytic activity after incubation with different concentrations of salts (NaCl, KCl, and LiCl) for 30 days, confirming it to be a halostable enzyme. It is the highest reported production of a recombinant endoglucanase, further increasing its potential for industrial applications.