Cell-free synthesis has been adopted in the bioconversion process due to its known advantages, such as fast production rate, high product content, and no substrate/product inhibition effect. In this study, the cell-free supernatant of Pseudomonas aeruginosa was used to improve the production of 7,10‐dihydroxy‐8(E)‐octadecenoic acid (DOD) from oleic acid. DOD production using cell-free supernatant demonstrated reduction in bioconversion duration and higher product concentration than conventional method using whole cell culture. The maximum DOD concentration (6.41 g/L) was obtained after 36 h of biotransformation using 1 % v/v oleic acid as a substrate with a productivity of 0.178 g/L/h and a yield of 74.8 %. DOD concentration, productivity, and yield using cell-free supernatant were 2.12, 7.12, and 2.22 times higher, respectively, than using the conventional whole cell culture method. Of the carbon and nitrogen sources used in pre-culture, galactose and sodium glutamate along with diammonium phosphate were found to be the most effective for DOD production. An incubation temperature of 27 °C and pH 8.0 were found to be most favorable for DOD production. In addition, sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis demonstrated the presence of enzymes related to DOD production in the cell-free supernatant, which was substantiated by performing DOD production experiment using the supernatant enzymes extracted from protein gel bands with oleic acid as a substrate. To the best of our knowledge, this is the first report on DOD production using a cell-free supernatant and verifying the existence of the relevant enzymes in the cell-free supernatant. Compared to whole cell process, cell-free DOD production holds several advantages, including higher DOD productivity which could be beneficial for large-scale production.

无细胞合成由于其众所周知的优点，如生产速度快、产物含量高、无底物/产物抑制作用，已在生物转化过程中采用。在本研究中，铜绿假单胞菌的无细胞上清液用于提高 7,10-二羟基-8( E)-十八碳烯酸 (DOD) 来自油酸。与使用全细胞培养的传统方法相比，使用无细胞上清液生产 DOD 表明生物转化持续时间缩短，产品浓度更高。使用 1% v/v 油酸作为底物进行生物转化 36 小时后获得最大 DOD 浓度 (6.41 g/L)，生产率为 0.178 g/L/h，产率为 74.8%。使用无细胞上清液的 DOD 浓度、生产力和产量分别比使用传统全细胞培养方法高 2.12、7.12 和 2.22 倍。在预培养中使用的碳源和氮源中，发现半乳糖和谷氨酸钠以及磷酸二铵对 DOD 生产最有效。发现 27 °C 的孵育温度和 pH 值 8.0 最有利于 DOD 生产。此外，十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析表明无细胞上清液中存在与 DOD 产生相关的酶，通过使用从蛋白质凝胶条带中提取的以油酸为底物的上清液酶进行 DOD 产生实验证实了这一点。据我们所知，这是使用无细胞上清液生产 DOD 并验证无细胞上清液中相关酶的存在的第一份报告。与全细胞工艺相比，无细胞 DOD 生产具有几个优势，包括更高的 DOD 生产率，这可能有利于大规模生产。以油酸为底物，从蛋白凝胶条带中提取的上清酶进行DOD生产实验证实了这一点。据我们所知，这是使用无细胞上清液生产 DOD 并验证无细胞上清液中相关酶的存在的第一份报告。与全细胞工艺相比，无细胞 DOD 生产具有几个优势，包括更高的 DOD 生产率，这可能有利于大规模生产。以油酸为底物，从蛋白凝胶条带中提取的上清酶进行DOD生产实验证实了这一点。据我们所知，这是使用无细胞上清液生产 DOD 并验证无细胞上清液中相关酶的存在的第一份报告。与全细胞工艺相比，无细胞 DOD 生产具有几个优势，包括更高的 DOD 生产率，这可能有利于大规模生产。