Abstract
Our previous research demonstrated phenylethyl alcohol was the key aroma in fermented chili peppers, identifying Pichia fermentans as the primary strain responsible for its production. However, the regulatory mechanisms underlying its production remained unclear. To address this, adaptive laboratory evolution against NaCl was utilized in this study to enhance the production of phenethyl and reveal the mechanism of metabolic regulatory of phenethyl alcohol in Pichia fermentans. Under high NaCl stress, the evolved strains increased phenylethyl alcohol production capacity by up to 64.01%. The improved NaCl tolerance of the strains contributes to the higher phenylethyl alcohol yield, primarily because phenylethyl alcohol, like NaCl, causes high osmotic pressure on the strain. The strain responds to NaCl stress by altering cell membrane components and related transport proteins through relevant gene mutations. Additionally, it enhances phenylethyl alcohol production efficiency by upregulating key enzyme-encoding genes (ARO8 and ADHs) in the Ehrlich pathway.
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