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Evaluation of Brachypodium distachyon L-Tyrosine Decarboxylase Using L-Tyrosine Over-Producing Saccharomyces cerevisiae.

Noda S, Shirai T, Mochida K, Matsuda F, Oyama S, Okamoto M, Kondo A - PLoS ONE (2015)

Bottom Line: It is suggested that two TDCs encoded in the transcripts Bradi2g51120.1 and Bradi2g51170.1 have L-tyrosine decarboxylation activity.The amount of tyramine produced by the resulting transformant was 6.6-fold higher (approximately 200 mg/L) than the control strain, indicating that B. distachyon TDC effectively converts L-tyrosine to tyramine.Our results suggest that B. distachyon possesses enzymes that are capable of modifying aromatic residues, and that S. cerevisiae is a suitable host for the production of L-tyrosine derivatives.

View Article: PubMed Central - PubMed

Affiliation: Biomass Engineering Program, RIKEN, Yokohama, Kanagawa, Japan.

ABSTRACT
To demonstrate that herbaceous biomass is a versatile gene resource, we focused on the model plant Brachypodium distachyon, and screened the B. distachyon for homologs of tyrosine decarboxylase (TDC), which is involved in the modification of aromatic compounds. A total of 5 candidate genes were identified in cDNA libraries of B. distachyon and were introduced into Saccharomyces cerevisiae to evaluate TDC expression and tyramine production. It is suggested that two TDCs encoded in the transcripts Bradi2g51120.1 and Bradi2g51170.1 have L-tyrosine decarboxylation activity. Bradi2g51170.1 was introduced into the L-tyrosine over-producing strain of S. cerevisiae that was constructed by the introduction of mutant genes that promote deregulated feedback inhibition. The amount of tyramine produced by the resulting transformant was 6.6-fold higher (approximately 200 mg/L) than the control strain, indicating that B. distachyon TDC effectively converts L-tyrosine to tyramine. Our results suggest that B. distachyon possesses enzymes that are capable of modifying aromatic residues, and that S. cerevisiae is a suitable host for the production of L-tyrosine derivatives.

No MeSH data available.


Related in: MedlinePlus

Culture profiles of transformants in SD medium containing 2% glucose as the carbon source.Time-courses of (A) cell growth, (B) glucose consumption, (C) ethanol production, and (D) tyramine production for YPH499/δU/δL/tdc70 (crosses), YPH499/δU/δLARO7fbr/tdc70 (triangles), YPH499/δUARO4fbr/δL/tdc70 (squares), and YPH499/δUARO4fbr/δLARO7fbr/tdc70 (circles). Each data point shows the average of 3 independent experiments, and error bars represent the standard deviation.
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pone.0125488.g003: Culture profiles of transformants in SD medium containing 2% glucose as the carbon source.Time-courses of (A) cell growth, (B) glucose consumption, (C) ethanol production, and (D) tyramine production for YPH499/δU/δL/tdc70 (crosses), YPH499/δU/δLARO7fbr/tdc70 (triangles), YPH499/δUARO4fbr/δL/tdc70 (squares), and YPH499/δUARO4fbr/δLARO7fbr/tdc70 (circles). Each data point shows the average of 3 independent experiments, and error bars represent the standard deviation.

Mentions: To evaluate the ability of B. distachyon TDC to convert L-tyrosine to tyramine, the gene encoding TDC70 was introduced into strains YPH499/δU/δL, YPH499/δU/δLARO7fbr, YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr. Fig 3(A) shows the time courses of cell growth of each transformant. Although the cell growth rates of YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr were higher than those of YPH499/δU/δL and YPH499/δU/δLARO7fbr, the maximal level of cell growth was similar among the four transformants. Fig 3(B) and 3(C) show time courses of the glucose consumption and ethanol production rates, respectively, of each transformant. The rates of glucose consumption and ethanol production of YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr were higher than those of YPH499/δU/δL and YPH499/δU/δLARO7fbr. Fig 3(D) shows the time courses of tyramine production by the recombinant strains. The maximal levels of tyramine production, which started after 12 h cultivation, reached by YPH499/δU/δL, YPH499/δU/δLARO7fbr, YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr were 30.4, 44.7, 113, and 200 mg/L, respectively, after 72 h of cultivation.


Evaluation of Brachypodium distachyon L-Tyrosine Decarboxylase Using L-Tyrosine Over-Producing Saccharomyces cerevisiae.

Noda S, Shirai T, Mochida K, Matsuda F, Oyama S, Okamoto M, Kondo A - PLoS ONE (2015)

Culture profiles of transformants in SD medium containing 2% glucose as the carbon source.Time-courses of (A) cell growth, (B) glucose consumption, (C) ethanol production, and (D) tyramine production for YPH499/δU/δL/tdc70 (crosses), YPH499/δU/δLARO7fbr/tdc70 (triangles), YPH499/δUARO4fbr/δL/tdc70 (squares), and YPH499/δUARO4fbr/δLARO7fbr/tdc70 (circles). Each data point shows the average of 3 independent experiments, and error bars represent the standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440718&req=5

pone.0125488.g003: Culture profiles of transformants in SD medium containing 2% glucose as the carbon source.Time-courses of (A) cell growth, (B) glucose consumption, (C) ethanol production, and (D) tyramine production for YPH499/δU/δL/tdc70 (crosses), YPH499/δU/δLARO7fbr/tdc70 (triangles), YPH499/δUARO4fbr/δL/tdc70 (squares), and YPH499/δUARO4fbr/δLARO7fbr/tdc70 (circles). Each data point shows the average of 3 independent experiments, and error bars represent the standard deviation.
Mentions: To evaluate the ability of B. distachyon TDC to convert L-tyrosine to tyramine, the gene encoding TDC70 was introduced into strains YPH499/δU/δL, YPH499/δU/δLARO7fbr, YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr. Fig 3(A) shows the time courses of cell growth of each transformant. Although the cell growth rates of YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr were higher than those of YPH499/δU/δL and YPH499/δU/δLARO7fbr, the maximal level of cell growth was similar among the four transformants. Fig 3(B) and 3(C) show time courses of the glucose consumption and ethanol production rates, respectively, of each transformant. The rates of glucose consumption and ethanol production of YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr were higher than those of YPH499/δU/δL and YPH499/δU/δLARO7fbr. Fig 3(D) shows the time courses of tyramine production by the recombinant strains. The maximal levels of tyramine production, which started after 12 h cultivation, reached by YPH499/δU/δL, YPH499/δU/δLARO7fbr, YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr were 30.4, 44.7, 113, and 200 mg/L, respectively, after 72 h of cultivation.

Bottom Line: It is suggested that two TDCs encoded in the transcripts Bradi2g51120.1 and Bradi2g51170.1 have L-tyrosine decarboxylation activity.The amount of tyramine produced by the resulting transformant was 6.6-fold higher (approximately 200 mg/L) than the control strain, indicating that B. distachyon TDC effectively converts L-tyrosine to tyramine.Our results suggest that B. distachyon possesses enzymes that are capable of modifying aromatic residues, and that S. cerevisiae is a suitable host for the production of L-tyrosine derivatives.

View Article: PubMed Central - PubMed

Affiliation: Biomass Engineering Program, RIKEN, Yokohama, Kanagawa, Japan.

ABSTRACT
To demonstrate that herbaceous biomass is a versatile gene resource, we focused on the model plant Brachypodium distachyon, and screened the B. distachyon for homologs of tyrosine decarboxylase (TDC), which is involved in the modification of aromatic compounds. A total of 5 candidate genes were identified in cDNA libraries of B. distachyon and were introduced into Saccharomyces cerevisiae to evaluate TDC expression and tyramine production. It is suggested that two TDCs encoded in the transcripts Bradi2g51120.1 and Bradi2g51170.1 have L-tyrosine decarboxylation activity. Bradi2g51170.1 was introduced into the L-tyrosine over-producing strain of S. cerevisiae that was constructed by the introduction of mutant genes that promote deregulated feedback inhibition. The amount of tyramine produced by the resulting transformant was 6.6-fold higher (approximately 200 mg/L) than the control strain, indicating that B. distachyon TDC effectively converts L-tyrosine to tyramine. Our results suggest that B. distachyon possesses enzymes that are capable of modifying aromatic residues, and that S. cerevisiae is a suitable host for the production of L-tyrosine derivatives.

No MeSH data available.


Related in: MedlinePlus