Significant enhancement of methionol production by co-expression of the aminotransferase gene ARO8 and the decarboxylase gene ARO10 in Saccharomyces cerevisiae.
Methionol is an important volatile sulfur flavor compound, which can be produced via the Ehrlich pathway in Saccharomyces cerevisiae. Aminotransferase and decarboxylase are essential enzymes catalyzing methionol biosynthesis. In this work, two aminotransferase genes ARO8 and ARO9 and one decarboxylase gene ARO10 were introduced into S. cerevisiae S288c, respectively, via an expression vector. Over-expression of ARO8 resulted in higher aminotransferase activity than that of ARO9. And the cellular decarboxylase activity was remarkably increased by over-expression of ARO10. A co-expression vector carrying both ARO8 and ARO10 was further constructed to generate the recombinant strain S810. Shaking flask experiments showed that the methionol yield from S810 reached 1.27 g L(-1), which was increased by 51.8 and 68.8% compared to that from the wild-type strain and the control strain harboring the empty vector. The fed-batch fermentation by strain S810 produced 3.24 g L(-1) of methionol after 72 h of cultivation in a bioreactor. These results demonstrated that co-expression of ARO8 and ARO10 significantly boosted the methionol production. It is the first time that more than 3.0 g L(-1) of methionol produced by genetically engineered yeast strain was reported by co-expression of the aminotransferase and decarboxylase via the Ehrlich pathway.
School of Food and Chemical Engineering, Beijing Technology & Business University, Beijing 100048, China Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology & Business University, Beijing 100048, China Beijing Key Laboratory of Flavor Chemistry, Beijing Technology & Business University, Beijing 100048, China.
Sheng
S
Yin
School of Food and Chemical Engineering, Beijing Technology & Business University, Beijing 100048, China Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology & Business University, Beijing 100048, China.
Tiandan
T
Lang
School of Food and Chemical Engineering, Beijing Technology & Business University, Beijing 100048, China Beijing Key Laboratory of Flavor Chemistry, Beijing Technology & Business University, Beijing 100048, China.
Xiao
X
Xiao
School of Food and Chemical Engineering, Beijing Technology & Business University, Beijing 100048, China Beijing Key Laboratory of Flavor Chemistry, Beijing Technology & Business University, Beijing 100048, China.
Li
L
Liu
School of Food and Chemical Engineering, Beijing Technology & Business University, Beijing 100048, China Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology & Business University, Beijing 100048, China Beijing Key Laboratory of Flavor Chemistry, Beijing Technology & Business University, Beijing 100048, China.
Baoguo
B
Sun
School of Food and Chemical Engineering, Beijing Technology & Business University, Beijing 100048, China Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology & Business University, Beijing 100048, China Beijing Key Laboratory of Flavor Chemistry, Beijing Technology & Business University, Beijing 100048, China wctbtbu@hotmail.com.
Chengtao
C
Wang
D002262:Carboxy-Lyases / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D005285:Fermentation; D015966:Gene Expression Regulation, Fungal; D005818:Genetic Engineering; D020005:Propanols / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D013379:Substrate Specificity; D013440:Sulfides / Q000378:metabolism*; D000637:Transaminases / Q000235:genetics* / Q000378:metabolism*
D020005:Propanols; D029701:Saccharomyces cerevisiae Proteins; D013440:Sulfides; D000637:Transaminases; D002262:Carboxy-Lyases; C555618:methionol
Saccharomyces cerevisiae; aminotransferase; co-expression; decarboxylase; methionol