Yin Sheng, Lang Tiandan, Xiao Xiao, Liu Li, Sun Baoguo, Wang Chengtao
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.
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.
FEMS Microbiol Lett. 2015 Mar;362(5). doi: 10.1093/femsle/fnu043. Epub 2014 Dec 8.
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.
甲醇是一种重要的挥发性含硫风味化合物,可通过酿酒酵母中的艾氏途径产生。转氨酶和脱羧酶是催化甲醇生物合成的关键酶。在本研究中,通过表达载体将两个转氨酶基因ARO8和ARO9以及一个脱羧酶基因ARO10分别导入酿酒酵母S288c中。ARO8的过表达导致转氨酶活性高于ARO9。ARO10的过表达显著提高了细胞脱羧酶活性。进一步构建了携带ARO8和ARO10的共表达载体,以产生重组菌株S810。摇瓶实验表明,S810的甲醇产量达到1.27 g L(-1),与野生型菌株和携带空载体的对照菌株相比,分别提高了51.8%和68.8%。S810菌株在生物反应器中培养72 h后,补料分批发酵产生了3.24 g L(-1)的甲醇。这些结果表明,ARO8和ARO10的共表达显著提高了甲醇产量。这是首次通过艾氏途径共表达转氨酶和脱羧酶,报道基因工程酵母菌株产生超过3.0 g L(-1)甲醇的情况。