A proteomic analysis of ferulic acid metabolism in Amycolatopsis sp. ATCC 39116

Applied Microbiology and Biotechnology
Florian MeyerA Steinbüchel

Abstract

The pseudonocardiate Amycolatopsis sp. ATCC 39116 is used for the biotechnical production of natural vanillin from ferulic acid. Our laboratory has performed genetic modifications of this strain previously, but there are still many gaps in our knowledge regarding its vanillin tolerance and the general metabolism. We performed cultivations with this bacterium and compared the proteomes of stationary phase cells before ferulic acid feeding with those during ferulic acid feeding. Thereby, we identified 143 differently expressed proteins. Deletion mutants were constructed and characterized to analyze the function of nine corresponding genes. Using these mutants, we identified an active ferulic acid β-oxidation pathway and the enzymes which constitute this pathway. A combined deletion mutant in which the β-oxidation as well as non-β-oxidation pathways of ferulic acid degradation were deleted was unable to grow on ferulic acid as the sole source of carbon and energy. This mutant differs from the single deletion mutants and was unable to grow on ferulic acid. Furthermore, we showed that the non-β-oxidation pathway is involved in caffeic acid degradation; however, its deletion is complemented even in the double deletion mutant. This sh...Continue Reading

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Citations

May 14, 2020·Applied Biochemistry and Biotechnology·Victor Contreras-JácquezAli Asaff-Torres

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Methods Mentioned

BETA
PCR
electrophoresis
protein folding

Software Mentioned

Phusion Tm
InterPro
Gen5
MASCOT
Chromeleon
Delta2D
AMIDS
JVirGel

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