An efficient transformation method for Bacillus subtilis DB104.
Summary of "An efficient transformation method for Bacillus subtilis DB104."
Bacillus subtilis strains are used for extracellular expression of enzymes (i.e., proteases, lipases, and cellulases) which are often engineered by directed evolution for industrial applications. B. subtilis DB104 represents an attractive directed evolution host since it has a low proteolytic activity and efficient secretion. B. subtilis DB104 is hampered like many other Bacillus strains by insufficient transformation efficiencies (≤10(3) transformants/μg DNA). After investigating five physical and chemical transformation protocols, a novel natural competent transformation protocol was established for B. subtilis DB104 by optimizing growth conditions and histidine concentration during competence development, implementing an additional incubation step in the competence development phase and a recovery step during the transformation procedure. In addition, the influence of the amount and size of the transformed plasmid DNA on transformation efficiency was investigated. The natural competence protocol is "easy" in handling and allows for the first time to generate large libraries (1.5 × 10(5) transformants/μg plasmid DNA) in B. subtilis DB104 without requiring microgram amounts of DNA.
Lehrstuhl für Biotechnologie, RWTH Aachen University, Worringerweg 1, 52074, Aachen, Germany.
This article was published in the following journal.
Name: Applied microbiology and biotechnology
- PubMed Source: http://www.ncbi.nlm.nih.gov/pubmed/22395911
- DOI: http://dx.doi.org/10.1007/s00253-012-3987-2
Medical and Biotech [MESH] Definitions
A family of SERINE ENDOPEPTIDASES isolated from Bacillus subtilis. EC 3.4.21.-
A species of gram-positive bacteria that is a common soil and water saprophyte.
A cyclic polypeptide antibiotic isolated from culture filtrates of Bacillus subtilis that acts as an antifungal agent.
A serine endopeptidase isolated from Bacillus subtilis. It hydrolyzes proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. It also hydrolyzes peptide amides. (From Enzyme Nomenclature, 1992) EC 22.214.171.124.
A complex of cyclic peptide antibiotics produced by the Tracy-I strain of Bacillus subtilis. The commercial preparation is a mixture of at least nine bacitracins with bacitracin A as the major constituent. It is used topically to treat open infections such as infected eczema and infected dermal ulcers. (From Goodman and Gilman, The Pharmacological Basis of Therapeutics, 8th ed, p1140)
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