Enzyme for synthesizing hydroxyl acetaldehyde and/or 1,3-dihydroxyacetone by catalyzing formaldehyde, and applications thereof
AssigneeTianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences
InventorsHuifeng Jiang, Xiaoyun Lu, Yuwan Liu, Yiqun Yang, Lina Lu, Sheng Yang, Junran Gou
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Abstract
An enzyme synthesizes hydroxyl acetaldehyde and/or 1,3-dihydroxyacetone by catalyzing formaldehyde. Site-directed mutation of benzoylformate decarboxylase (BFD) creates a mutant of the enzyme, which can polymerize the formaldehyde, A phosphoketalose (F/XPK) generates acetyl phosphoric acid from the hydroxyl acetaldehyde or 1,3-dihydroxyacetone (DHA). Combination with phosphotransacetylase (Pta) provides a route from the formaldehyde to acetyl coenzyme A in three steps.
Claims15
A method for preparing hydroxyl acetaldehyde and/or 1,3-dihydroxyacetone, comprising:
- catalyzing formaldehyde to be condensed to hydroxyl acetaldehyde and/or 1,3-dihydroxyacetone with a benzoylformate decarboxylase (BFD) mutant protein using formaldehyde as a substrate,
- wherein the BFD mutant protein comprises the amino acid sequence of SEQ ID NO:
- 1 with the exception of the mutations W86R and N87T, and optionally one or more mutations selected from the group consisting of L109G, L110E, H281V, Q282F, T377M, T380C, T380Y, and A460M, wherein the amino acid number corresponds to the amino acid sequence of SEQ ID NO:
- 1.
A method for producing acetyl phosphoric acid using formaldehyde, comprising:
- (i) catalyzing formaldehyde to be condensed to hydroxyl acetaldehyde and/or 1,3-dihydroxyacetone with a BFD mutant protein, using formaldehyde as a substrate;
- (ii) using the hydroxyl acetaldehyde and/or 1, 3-dihydroxyacetone obtained in (i) as a substrate to prepare acetyl phosphoric acid,
- wherein the BFD mutant protein comprises the amino acid sequence of SEQ ID NO:
- 1 with the exception of the mutations W86R and N87T, and optionally one or more mutations selected from the group consisting of L109G, L110E, H281V, Q282F, T377M, T380C, T380Y, and A460M, wherein the amino acid number corresponds to the amino acid sequence of SEQ ID NO:1.
A method for producing acetyl coenzyme A using formaldehyde, the method comprising:
- (i) catalyzing formaldehyde to be condensed to hydroxyl acetaldehyde and/or 1,3-dihydroxyacetone with a BFD mutant protein, using formaldehyde as a substrate;
- (ii) using hydroxyl acetaldehyde and/or 1,3-dihydroxyacetone obtained in (i) as a substrate to prepare acetyl phosphoric acid;
- and (iii) using the acetyl phosphoric acid as a substrate to prepare acetyl coenzyme A,
- wherein the BFD mutant protein comprises the amino acid sequence of SEQ ID NO:
- 1 with the exception of the mutations W86R and N87T, and optionally one or more mutations selected from the group consisting of L109G, L110E, H281V, Q282F, T377M, T380C, T380Y, and A460M, wherein the amino acid number corresponds to the amino acid sequence of SEQ ID NO:1.
Dependencies are inferred from the claim text.