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Literature summary for 1.2.1.67 extracted from

  • Fleige, C.; Meyer, F.; Steinbuechel, A.
    Metabolic engineering of the actinomycete Amycolatopsis sp. strain ATCC 39116 towards enhanced production of natural vanillin (2016), Appl. Environ. Microbiol., 82, 3410-3419 .
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
synthesis the Gram-positive bacterium Amycolatopsis sp. strain ATCC 39116 is used for the fermentative production of natural vanillin from ferulic acid on an industrial scale. The strain is known for its outstanding tolerance to this toxic product vanillin Amycolatopsis sp. ATCC 39116

Cloned(Commentary)

Cloned (Comment) Organism
gene vdh Amycolatopsis sp. ATCC 39116

Protein Variants

Protein Variants Comment Organism
additional information metabolic engineering of the actinomycete Amycolatopsis sp. strain ATCC 39116 towards enhanced production of natural vanillin. Degradation of vanillin is decreased by more than 90% through deletion of the vdh gene, which codes for the central vanillin catabolism enzyme, vanillin dehydrogenase. This mutation results in improvement of the final concentration of vanillin by more than 2.2 g/liter, with a molar yield of 80.9%. Further improvement is achieved with constitutive expression of the vanillin anabolism genes ech and fcs, coding for the enzymes feruloyl-coenzyme A (CoA) synthetase (fcs) and enoyl-CoA hydratase/aldolase (ech). The transcription of both genes is shown to be induced by ferulic acid, which explains the unwanted adaptation phase in the fermentation process before vanillin is efficiently produced by the wild-type cells. Through the constitutive and enhanced expression of the two genes, the adaptation phase is eliminated and a final vanillin concentration of 19.3 g/liter, with a molar yield of 94.9%, is obtained. Moreover, an even higher final vanillin concentration of 22.3 g/liter is achieved, at the expense of a lower molar yield, by using an improved feeding strategy. The vanillin is produced almost without by-products, with a molar yield that nearly approaches the theoretical maximum. Generation of a vdh deletion mutant with replacement of vdh gene by kanamycin resistance gene, deletion of vdh via homologous recombination Amycolatopsis sp. ATCC 39116

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
vanillin + NAD+ + H2O Amycolatopsis sp. ATCC 39116
-
vanillate + NADH + 2 H+
-
?
vanillin + NAD+ + H2O Amycolatopsis sp. ATCC 39116 75iv2
-
vanillate + NADH + 2 H+
-
?

Organism

Organism UniProt Comment Textmining
Amycolatopsis sp. ATCC 39116 K9UV87
-
-
Amycolatopsis sp. ATCC 39116 75iv2 K9UV87
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
vanillin + NAD+ + H2O
-
Amycolatopsis sp. ATCC 39116 vanillate + NADH + 2 H+
-
?
vanillin + NAD+ + H2O
-
Amycolatopsis sp. ATCC 39116 75iv2 vanillate + NADH + 2 H+
-
?

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Amycolatopsis sp. ATCC 39116

General Information

General Information Comment Organism
malfunction a gene vdh deletion mutant of Amycolatopsis sp. strain ATCC 39116 shows increased levels of accumulated vanillin Amycolatopsis sp. ATCC 39116