1,3-dicarbonyl-dioxygenase - a new c-c-bond cleaving enzyme
AssigneeBioCatalytics, Inc.
InventorsGrit Straganz, Anton Glieder, Lothar Brecker
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Abstract
An enzyme that cleaves 1,3-dicarbonyl compounds by incorporating molecular oxygen into the substrate in amounts equimolar to the amount of substrate cleaved. The enzyme from Acinetobacter johnsonii is a multimer of about 67 kilodaltons made up of subunits of about 16.6 kilodaltons. Nucleotide and amino acid sequence analysis of a cloned A. johnsonii subunit indicates little homology with other proteins. Native or recombinant enzyme can be used to decontaminate or detoxify acetylacetone or related diketones.
Claims32
A purified enzyme exhibiting 1,3-dicarbonyl-dioxygenase activity.
An isolated polypeptide subunit of an enzyme exhibiting 1,3-dicarbonyl-dioxygenase activity.
An isolated polynucleotide encoding a subunit of an enzyme exhibiting 1,3-dicarbonyl-dioxygenase activity, said polynucleotide comprising a nucleotide sequence selected from the group consisting of:
- (a) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:
- 2;
- (b) a nucleotide sequence encoding an amino acid sequence that has at least 70% identity to SEQ ID NO:
- 2;
- (c) the nucleotide sequence set forth in SEQ ID NO:
- 1;
- (d) a nucleotide sequence having at least 60% identity to SEQ ID NO:
- 1;
- (e) the nucleotide sequence set forth in SEQ ID NO:
- 3;
- (f) the nucleotide sequence set forth in SEQ ID NO:
- 4;
- (g) the nucleotide sequence set forth in SEQ ID NO:
- 8;
- (h) the nucleotide sequence set forth in SEQ ID NO:
- 9;
- (i) the nucleotide sequence set forth in SEQ ID NO:
- 10;
- (j) the nucleotide sequence set forth in SEQ ID NO:
- 11;
- and
- (k) a nucleotide sequence that is complementary to a nucleotide sequence of (a)-(j).
A process for producing a subunit of an enzyme exhibiting 1,3-dicarbonyl-dioxygenase activity, the process comprising the steps of:
- (a) culturing a cell expressing the subunit, the culturing occurring under conditions sufficient for the cell to produce the subunit, and
- (b) isolating the subunit from the cell culture.
A process for producing an enzyme exhibiting 1,3-dicarbonyl-dioxygenase activity, the process comprising the steps of:
- (a) culturing a cell expressing the enzyme, the culturing occurring under conditions sufficient for the cell to produce the enzyme, and
- (b) isolating the enzyme from the cell culture.
A composition comprising an enzyme exhibiting 1,3-dicarbonyl-dioxygenase activity.
A method of cleaving a 1,3-dicarbonyl compound comprising reacting the compound in the presence of O2 with an enzyme exhibiting 1,3-dicarbonyl-dioxygenase activity.
Dependencies are inferred from the claim text.