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- Title
Production and characteristics of the recombinant extracellular bifunctional endoglucanase of the polyextremophilic bacterium Bacillus halodurans and its applicability in saccharifying agro-residues.
- Authors
Prabhu, Ranjitha; Parashar, Deepak; Satyanarayana, T.
- Abstract
The recombinant alkalistable and moderately thermostable bifunctional endoglucanase gene ( BhCell- Xyl) of polyextremophilic bacterium Bacillus halodurans TSLV1 has been expressed in Pichia pastoris under constitutive GAP as well as inducible AOX promoters. A higher titre of recombinant BhCell-Xyl was attained after induction (4.8 U mL) as compared to that of the constitutive production (2.1 U mL). The recombinant P. pastoris strains integrated two copies of BhCell- Xyl under AOX and GAP promoters. The pure recombinant BhCell-Xyl is a glycoprotein of 66 kDa, which is optimally active at 60 °C and pH 6.0 and 8.0. Glycosylated BhCell-Xyl exhibits higher thermostability than that of the native enzyme. The analysis of amino acids of BhCell-Xyl revealed that multiple factors are responsible for its thermostability. Kinetics and in silico analysis of the enzyme suggested that BhCell-Xyl has one active site for both endocellulase and endoxylanase activities. The BhCell-Xyl possesses a carbohydrate binding domain and saccharifies lignocellulosic agro-residues to xylo-oligosaccharides and cello-oligosaccharides, suggesting its potential application in generating fermentable sugars from renewable agro-residues for biofuel and fine chemical industries.
- Subjects
BACILLUS halodurans; PICHIA pastoris; GLYCOPROTEINS; AMINO acids; LIGNOCELLULOSE; CARBOHYDRATES; DYNAMICS
- Publication
Bioprocess & Biosystems Engineering, 2017, Vol 40, Issue 5, p651
- ISSN
1615-7591
- Publication type
Article
- DOI
10.1007/s00449-016-1730-6