A Broad Spectrum Racemase in Pseudomonas putida KT2440 Plays a Key Role in Amino Acid Catabolism

Frontiers in Microbiology
Atanas D Radkov, Luke A Moe

Abstract

The broad-spectrum amino acid racemase (Alr) of Pseudomonas putida KT2440 preferentially interconverts the l- and d-stereoisomers of Lys and Arg. Despite conservation of broad-spectrum racemases among bacteria, little is known regarding their physiological role. Here we explore potential functional roles for Alr in P. putida KT2440. We demonstrate through cellular fractionation that Alr enzymatic activity is found in the periplasm, consistent with its putative periplasm targeting sequence. Specific activity of Alr is highest during exponential growth, and this activity corresponds with an increased accumulation of d-Lys in the growth medium. An alr gene knockout strain (Δalr) was generated and used to assess potential roles for the alr gene in peptidoglycan structure, producing soluble signaling compounds, and amino acid metabolism. The stationary phase peptidoglycan structure did not differ between wild-type and Δalr strains, indicating that products resulting from Alr activity are not incorporated into peptidoglycan under these conditions. RNA-seq was used to assess differences in the transcriptome between the wild-type and Δalr strains. Genes undergoing differential expression were limited to those involved in amino acid met...Continue Reading

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Citations

Jul 22, 2019·Applied and Environmental Microbiology·Andrew D SteenMarc J Alperin
Apr 9, 2021·Environmental Microbiology·Alena AliashkevichFelipe Cava
Apr 1, 2021·Journal of Biochemistry·Tetsuya Miyamoto, Hiroshi Homma

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Datasets Mentioned

BETA
PRJNA471301

Methods Mentioned

BETA
PCR
RNA-seq

Software Mentioned

CLC Genomics Workbench

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