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OwnerJuan Luis Ramos, Arie Ben-Bassat +3inventors
InventorsJuan Luis Ramos, Arie Ben-Bassat, Estrella Duque, Patricia Godoy, Marie Isable Ramos-Gonzalez
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This invention relates to the isolation of a novel tonB operon from Pseudomonas putida. These genes are useful to render the cells more sensitive to antibiotics, toluene, pHBA, aromatic compounds, parabenes, and aromatic amino acids after inactivation with specific mutant allels or more tolerant to these compounds after overexpression with appropriate expression vector. These findings are important in the field of medicine and biotechnology and biocatalysis. In addition a screen to identify pHBA tolerant genes is provided and strains with significant tolerance to pHBA were identified. These strains are important for pHBA production.
An isolated nucleic acid fragment of the tonB operon from Pseudomonas selected from the group consisting of:
An isolated nucleic acid fragment encoding ExbB from Pseudomonas selected from the group consisting of:
An isolated nucleic acid fragment comprising 1) a first nucleotide sequence encoding a polypeptide of amino acids that has at least 90% identity based on Xnr BLAST algorithm, when compared to a polypeptide encoded by the Pseudomonas putida WCS ExbB gene (Ac. Number X70139), or 2) a second nucleotide sequence comprising the complement of the first nucleotide sequence.
An isolated nucleic acid fragment encoding exbD from Pseudomonas selected from the group consisting of:
An isolated nucleic acid fragment comprising 1) a first nucleotide sequence encoding a polypeptide of amino acids that has at least 97.9% identity based on Xnr BLAST algorithm when compared to a polypeptide encoded by the Pseudomonas putida WCS exbD gene (Ac. Number X70139) or 2) a second nucleotide sequence comprising the complement of the first nucleotide sequence.
An isolated nucleic acid fragment encoding TonB from Pseudomonas selected from the group consisting of:
An isolated nucleic acid fragment comprising 1) a first nucleotide sequence encoding a polypeptide of amino acids that has at least 93.8% identity based on Xnr BLAST algorithm, when compared to a polypeptide encoded by the Pseudomonas putida WCS tonB gene (Ac. Number X70139), or 2) a second nucleotide sequence comprising the complement of the first nucleotide sequence.
A transformed host bacterial cell selected from the group consisting of:
An isolated nucleic acid fragment selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:
A method to produce pHBA comprising:
A method to obtain a bacterial strain more sensitive than the parent strain to antibiotics, aromatic carboxylic, parabens, and aromatic amino acids comprising:
A method to obtain a transformed bacterial strain that is more tolerant to pHBA and aromatic amino acids than an un-transformed host bacterial strain, comprising:
Dependencies are inferred from the claim text.