Limits...
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

Evaluation of L-tyrosine over-producing S. cerevisiae constructed in this study.Each bar chart shows the average of 3 independent experiments, and error bars represent the standard deviation. (A) Evaluation of L-tyrosine productivity in the culture supernatants of YPH499/δU/δL, YPH499/δUARO4fbr/δL, YPH499/δU/δLARO7fbr, and YPH499/δUARO4fbr/δLARO7fbr. (B) Determination of ARO7 and ARO7fbr gene copy numbers in YPH499/δU/δL, YPH499/δUARO4fbr/δL, YPH499/δU/δLARO7fbr, and YPH499/δUARO4fbr/δLARO7fbr (ARO4fbr; Ser to Ala substitution in ARO4 at position 195: Gly to Ser substitution in ARO7 at position 141).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4440718&req=5

pone.0125488.g002: Evaluation of L-tyrosine over-producing S. cerevisiae constructed in this study.Each bar chart shows the average of 3 independent experiments, and error bars represent the standard deviation. (A) Evaluation of L-tyrosine productivity in the culture supernatants of YPH499/δU/δL, YPH499/δUARO4fbr/δL, YPH499/δU/δLARO7fbr, and YPH499/δUARO4fbr/δLARO7fbr. (B) Determination of ARO7 and ARO7fbr gene copy numbers in YPH499/δU/δL, YPH499/δUARO4fbr/δL, YPH499/δU/δLARO7fbr, and YPH499/δUARO4fbr/δLARO7fbr (ARO4fbr; Ser to Ala substitution in ARO4 at position 195: Gly to Ser substitution in ARO7 at position 141).

Mentions: YPH499/δU/δL, YPH499/δU/δLARO7fbr, YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr were cultured in SD medium containing 2% glucose, and the culture supernatants were analyzed by GC-MS to quantify the amount of L-tyrosine produced after 72 h cultivation (Fig 2A). A total of 0.80, 1.99 and 4.84 mg/L tyrosine was found in the culture supernatant of YPH499/δU/δLARO7fbr, YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr, respectively, whereas no tyrosine was detected in the culture supernatant of the control strain YPH499/δU/δL. The copy number of ARO4fbr integrated into the genome of YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr, which originated from YPH499/δUARO4fbr/δL, was estimated by real-time PCR to be 2 in all strains. In contrast, the copy number of ARO7fbr in the genome of YPH499/δUARO4fbr/δLARO7fbr was estimated to be approximately 20, whereas that of YPH499/δU/δLARO7fbr was approximately 9 (Fig 2(B)).


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)

Evaluation of L-tyrosine over-producing S. cerevisiae constructed in this study.Each bar chart shows the average of 3 independent experiments, and error bars represent the standard deviation. (A) Evaluation of L-tyrosine productivity in the culture supernatants of YPH499/δU/δL, YPH499/δUARO4fbr/δL, YPH499/δU/δLARO7fbr, and YPH499/δUARO4fbr/δLARO7fbr. (B) Determination of ARO7 and ARO7fbr gene copy numbers in YPH499/δU/δL, YPH499/δUARO4fbr/δL, YPH499/δU/δLARO7fbr, and YPH499/δUARO4fbr/δLARO7fbr (ARO4fbr; Ser to Ala substitution in ARO4 at position 195: Gly to Ser substitution in ARO7 at position 141).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0125488.g002: Evaluation of L-tyrosine over-producing S. cerevisiae constructed in this study.Each bar chart shows the average of 3 independent experiments, and error bars represent the standard deviation. (A) Evaluation of L-tyrosine productivity in the culture supernatants of YPH499/δU/δL, YPH499/δUARO4fbr/δL, YPH499/δU/δLARO7fbr, and YPH499/δUARO4fbr/δLARO7fbr. (B) Determination of ARO7 and ARO7fbr gene copy numbers in YPH499/δU/δL, YPH499/δUARO4fbr/δL, YPH499/δU/δLARO7fbr, and YPH499/δUARO4fbr/δLARO7fbr (ARO4fbr; Ser to Ala substitution in ARO4 at position 195: Gly to Ser substitution in ARO7 at position 141).
Mentions: YPH499/δU/δL, YPH499/δU/δLARO7fbr, YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr were cultured in SD medium containing 2% glucose, and the culture supernatants were analyzed by GC-MS to quantify the amount of L-tyrosine produced after 72 h cultivation (Fig 2A). A total of 0.80, 1.99 and 4.84 mg/L tyrosine was found in the culture supernatant of YPH499/δU/δLARO7fbr, YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr, respectively, whereas no tyrosine was detected in the culture supernatant of the control strain YPH499/δU/δL. The copy number of ARO4fbr integrated into the genome of YPH499/δUARO4fbr/δL and YPH499/δUARO4fbr/δLARO7fbr, which originated from YPH499/δUARO4fbr/δL, was estimated by real-time PCR to be 2 in all strains. In contrast, the copy number of ARO7fbr in the genome of YPH499/δUARO4fbr/δLARO7fbr was estimated to be approximately 20, whereas that of YPH499/δU/δLARO7fbr was approximately 9 (Fig 2(B)).

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