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Engineering Bacillus megaterium for production of functional intracellular materials

Microbial Cell Factories. 2017; 
Katrin Grage , Paul McDermott and Bernd H. A. Rehm*
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Codon Optimization Te genes phaA, phaB and phaC from Cupriavidus necator plus the DNA sequences encoding the N- and C-terminally fused Z-domains were synthesized (GenScript) as codon optimized for expression in B. megaterium. Te ZZ/ZZC-ZZ encoding sequences were designed to translate into a protein with identical amino acid sequence compared to the E. coli produced protein [16]. phaC, phaA, phaB, the N-terminal Z domains and the C-terminal Z domains were all synthesized individually by GenScript and provided in plasmid pUC57. Get A Quote

摘要

Background: Over the last 10–15 years, a technology has been developed to engineer bacterial poly(3-hydroxybutyrate) (PHB) inclusions as functionalized beads, for applications such as vaccines, diagnostics and enzyme immobilization. This has been achieved by translational fusion of foreign proteins to the PHB synthase (PhaC). The respective fusion protein mediates self-assembly of PHB inclusions displaying the desired protein function. So far, beads have mainly been produced in recombinant Escherichia coli, which is problematic for some applications as the lipopolysaccharides (LPS) co-purifed with such inclusions are toxic to humans and animals. Results: In this study, we have bioengineered the formation of f... More

关键词

Poly(3-hydroxybutyrate) (PHB), PHA synthase, Bacillus megaterium, Endotoxin, Genetic engineering, Functionalized beads, ZZ-domain, IgG binding, Sporulation, ∆spoIIE