Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China.
ACS Synth Biol. 2021 Mar 19;10(3):495-504. doi: 10.1021/acssynbio.0c00446. Epub 2021 Feb 12.
Ethyl acetate can be synthesized from acetyl-CoA and ethanol a reaction by alcohol acetyltransferases (AATase) in yeast. In order to increase the yield of acetyl-CoA, different terminators were used to optimize the expressions of acetyl-CoA synthetase (ACS1/2) and aldehyde dehydrogenase (ALD6) to increase the contents of acetyl-CoA in . coding AATase was coexpressed in expression cassettes of ACS1/ACS2 and ALD6 to promote the carbon flux toward ethyl acetate from acetyl-CoA. Further to improve ethyl acetate production, four heterologous AATase including HuvEAT1 (), KamEAT1 (), VAAT (wild strawberry), and AeAT9 (kiwifruit) were introduced. Subsequently mitochondrial transport and utilization of pyruvate and acetyl-CoA were impeded to increase the ethyl acetate accumulation in cytoplasm. Under the optimal fermentation conditions, the engineered strain of PGAeΔPOR2 produced 1.69 g/L ethyl acetate, which was the highest value reported to date by metabolic engineering methods.
乙酸乙酯可以通过酵母中的醇乙酰基转移酶 (AATase) 从乙酰辅酶 A 和乙醇合成。为了增加乙酰辅酶 A 的产量，使用了不同的终止子来优化乙酰辅酶 A 合成酶 (ACS1/2) 和醛脱氢酶 (ALD6) 的表达，以增加中乙酰辅酶 A 的含量。编码 AATase 的基因与 ACS1/ACS2 和 ALD6 的表达盒共表达，以促进从乙酰辅酶 A 向乙酸乙酯的碳通量。为了进一步提高乙酸乙酯的产量，引入了四种异源 AATase，包括 HuvEAT1()、KamEAT1()、VAAT（野草莓）和 AeAT9（猕猴桃）。随后，通过抑制丙酮酸和乙酰辅酶 A 的线粒体转运和利用，增加了细胞质中乙酸乙酯的积累。在最佳发酵条件下，工程菌 PGAeΔPOR2 生产了 1.69 g/L 的乙酸乙酯，这是迄今为止通过代谢工程方法报道的最高值。
