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OwnerGerhard Miksch, Erwin Flaschel +3inventors
InventorsGerhard Miksch, Erwin Flaschel, Roland Breves, Karl-Heinz Maurer, sophia Kleist
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The invention relates to a method for producing recombinant proteins by gram-negative bacteria. According to the inventive method, the products are released into the surrounding medium, thereby allowing for high expression and production rates. To this end, the gene of the recombinant protein to be produced is placed under the control of a promoter derived from a gram-positive organism, preferably from a promoter derived from the genus Bacillus that in nature does not control said gene, and a system becomes active that partially opens the outer membrane of the bacteria produced. The preferred bacteria are E. coli or Klebsiella, promoters that are not necessarily inducible from outside, especially constitutive promoters such as the β-glucanase promoter of Bacillus amyloliquefaciens (bgl promoter) and the colicin system. The protein is thereby released into the surrounding medium from where it can be easily purified. The inventive method allows for making the fermentative production of protein more efficient. The inventive system is for example suitable for producing α-amylases or bacterial phytases.
A method for producing a recombinant protein by Gram-negative bacteria, which protein is at least partially secreted into the medium surrounding said bacteria with the aid of a system which partially opens the outer membrane of these bacteria, characterized in that the recombinant protein to be produced is expressed under the control of a promoter from a Gram-positive organism, preferably from an organism of the genus Bacillus, which promoter does not naturally regulate the corresponding gene or a gene highly homologous to this gene.
A secretion cassette which possesses the genetic elements responsible for the membrane-opening properties of the membrane-opening system, in particular the E. coli colicin system and/or a stationary-phase promoter, very particularly the gene for the Kil protein and/or an E. coli stationary-phase promoter including, in particular, the fic promoter.
A derivative of the microorganism deposited with the application number DSM 14225.
A method for fermentation of Gram-negative bacteria producing a recombinant protein which is at least partially secreted into the medium surrounding said bacteria with the aid of a system which partially opens the outer membrane of these bacteria, characterized in that the recombinant protein is expressed under the control of a promoter from a Gram-positive organism, preferably from an organism of the genus Bacillus, which promoter does not naturally regulate the corresponding gene or a gene highly homologous to this gene.
Dependencies are inferred from the claim text.