Journal
JOURNAL OF BIOTECHNOLOGY
Volume 146, Issue 3, Pages 100-104Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2010.02.006
Keywords
Thermobifida fusca; Carboxylesterase; High level expression; Hydrophobic proteins
Categories
Funding
- Sachsisches Staatsministerium fur Umwelt und Landwirtschaft [13-8811.61/215-1]
Ask authors/readers for more resources
The gram-positive thermophilic actinomycete Thermobifida fusca KW3 secretes a highly hydrophobic carboxylesterase (TfCa) that is able to hydrolyze poly(ethylene terephthalate). TfCa was produced in the Escherichia call strain BL21(DE3) as a fusion protein consisting of a pelB leader sequence to ensure periplasmic localization of the protein and a His(6) tag for use in its purification. To enhance the recombinant enzyme yield, the tfca gene from T. fusca KW3 was successfully optimized for codon usage in E. coli. In addition, the gene expression induction conditions were optimized and the temperature for cell cultivation was lowered to reduce inclusion body formation. The optimized codons and expression conditions yielded 4500-fold higher TfCa activity than the wild-type strain. Using a pH-controlled bioreactor for cultivation, a TfCa protein concentration of 41.6 mg/L was achieved. (C) 2010 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available