Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin

Fleige C, Meyer F, Steinbüchel A (2016)
Applied and Environmental Microbiology 82(11): 3410-3419.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Autor*in
Fleige, Christian; Meyer, FlorianUniBi ; Steinbüchel, Alexander
Abstract / Bemerkung
The Gram-positive bacterium Amycolatopsis sp. 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. In order to improve the productivity of the fermentation process, the strain's metabolism was engineered for higher final concentrations and molar yields. Degradation of vanillin could be decreased by more than 90% through deletion of the vdh gene, which codes for the central vanillin catabolism enzyme, vanillin dehydrogenase. This mutation resulted in improvement of the final concentration of vanillin by more than 2.2 g/liter, with a molar yield of 80.9%. Further improvement was 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 was shown to be induced by ferulic acid, which explains the unwanted adaptation phase in the fermentation process before vanillin was efficiently produced by the wild-type cells. Through the constitutive and enhanced expression of the two genes, the adaptation phase was eliminated and a final vanillin concentration of 19.3 g/liter, with a molar yield of 94.9%, was obtained. Moreover, an even higher final vanillin concentration of 22.3 g/liter was achieved, at the expense of a lower molar yield, by using an improved feeding strategy. This is the highest reported vanillin concentration reached in microbial fermentation processes without extraction of the product. Furthermore, the vanillin was produced almost without by-products, with a molar yield that nearly approached the theoretical maximum.
Erscheinungsjahr
2016
Zeitschriftentitel
Applied and Environmental Microbiology
Band
82
Ausgabe
11
Seite(n)
3410-3419
ISSN
0099-2240
eISSN
1098-5336
Page URI
https://pub.uni-bielefeld.de/record/2950265

Zitieren

Fleige C, Meyer F, Steinbüchel A. Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin. Applied and Environmental Microbiology. 2016;82(11):3410-3419.
Fleige, C., Meyer, F., & Steinbüchel, A. (2016). Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin. Applied and Environmental Microbiology, 82(11), 3410-3419. https://doi.org/10.1128/AEM.00802-16
Fleige, C., Meyer, F., and Steinbüchel, A. (2016). Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin. Applied and Environmental Microbiology 82, 3410-3419.
Fleige, C., Meyer, F., & Steinbüchel, A., 2016. Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin. Applied and Environmental Microbiology, 82(11), p 3410-3419.
C. Fleige, F. Meyer, and A. Steinbüchel, “Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin”, Applied and Environmental Microbiology, vol. 82, 2016, pp. 3410-3419.
Fleige, C., Meyer, F., Steinbüchel, A.: Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin. Applied and Environmental Microbiology. 82, 3410-3419 (2016).
Fleige, Christian, Meyer, Florian, and Steinbüchel, Alexander. “Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin”. Applied and Environmental Microbiology 82.11 (2016): 3410-3419.

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