Endoglucanase E, produced at high level in Escherichia coli as a lacZ′ fusion protein, is part of the Clostridium thermocellum cellulosome

1990 ◽  
Vol 12 (9) ◽  
pp. 656-662 ◽  
Author(s):  
Geoffrey P. Hazlewood ◽  
Keith Davidson ◽  
Jonathan H. Clarke ◽  
Alastair J. Durrant ◽  
Judith Hall ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Clostridium Thermocellum ◽  
Lacz Fusion ◽  
High Level

scholarly journals Soluble Production, Characterization, and Structural Aesthetics of an Industrially Important Thermostable β-Glucosidase from Clostridium thermocellum in Escherichia coli

2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Syed Shoaib Ahmed ◽  
Mohsina Akhter ◽  
Muhammad Sajjad ◽  
Roquyya Gul ◽  
Sana Khurshid
Keyword(s):  
Escherichia Coli ◽  
Clostridium Thermocellum ◽  
Expression System ◽  
Alpha Helix ◽  
Cellular Protein ◽  
Industrial Applications ◽  
Heat Inactivation ◽  
Potential Candidate ◽  
Cellulolytic Activity ◽  
High Level

This study aims to achieve high-level soluble expression and characterization of a thermostable industrially important enzyme, i.e., beta-glucosidase (BglA; EC: 3.2.1.21), from Clostridium thermocellum (C. thermocellum) by cloning in an Escherichia coli (E. coli) expression system. BglA was expressed as a partially soluble component of total cellular protein (TCP) having a molecular weight of ∼53 kDa with 50% of it as soluble fraction. Purification in two steps, namely, heat inactivation and Ni-chromatography, yielded approximately 30% and 15% of BglA, respectively. The purified (∼98%) BglA enzyme showed promising activity against the salicin substrate having a Km of 19.83 mM and a Vmax of 0.12 μmol/min. The enzyme had an optimal temperature and pH of 50°C and 7.0, respectively, while retaining its catalytic activity up till 60°C and at pH 7. The optimized maximum expression level was attained in M9NG medium with lactose as an inducer. Circular dichroism revealed presence of alpha helix (43.50%) and small percentage of beta sheets (10.60%). Factors like high-end cellulolytic activity, fair thermal stability, stability against low pH, and ease of purification make BglA from C. thermocellum a potential candidate in industrial applications.

High-level expression of tetanus toxin fragment C‒thioredoxin fusion protein in Escherichia coli

2000 ◽  
Vol 31 (2) ◽  
pp. 91 ◽  
Author(s):  
Adriana V. Ribas ◽  
Paulo L. Ho ◽  
Martha M. Tanizaki ◽  
Isaias Raw ◽  
Ana L.T.O. Nascimento
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Tetanus Toxin ◽  
High Level Expression ◽  
High Level

High-level soluble expression of hIGF-1 fusion protein in recombinant Escherichia coli

2010 ◽  
Vol 45 (8) ◽  
pp. 1401-1405 ◽  
Author(s):  
Danping Zhang ◽  
Peilian Wei ◽  
Limei Fan ◽  
Jiazhang Lian ◽  
Lei Huang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Soluble Expression ◽  
Recombinant Escherichia Coli ◽  
High Level
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