A first-of-its-kind, fully continuous synthesis of wax esters from biobased precursors (glucose, fatty acids) was developed using metabolically engineered cells and in vitro enzyme catalysis. The cells, overexpressing fatty acyl-CoA reductase and xylose reductase, could be immobilized onto polyesters and packed in a continuous reactor. The immobilized cells were employed in the bioconversion, incorporating in situ extraction using dodecane as the solvent. Such extractive bioconversion was capable of producing fatty alcohols continuously at a productivity of 8.2 mg/(L·h). The immiscible aqueous-dodecane flow stream from the extractive bioconversion was then separated using an in-line membrane-based separator. The dodecane-rich phase was directed into an enzymatic reactor containing Novozyme 435 for the esterification of fatty alchols and fatty acids into the wax esters. A continuous production of wax esters (6.38–23.35 mg/(L·h)) was achieved as a result of the successful streamlining of the cascade biocatalytic process.

中文翻译：

---

结合全细胞和体外生物催化的蜡酯集成连续流生产

利用代谢工程细胞和体外酶催化，开发了一种首创的从生物基前体（葡萄糖、脂肪酸）完全连续合成蜡酯的方法。过表达脂肪酰辅酶A还原酶和木糖还原酶的细胞可以固定在聚酯上并包装在连续反应器中。固定化细胞用于生物转化，并使用十二烷作为溶剂进行原位提取。这种萃取生物转化能够以8.2 mg/(L·h)的生产率连续生产脂肪醇。然后使用在线膜分离器分离来自萃取生物转化的不混溶的水性十二烷流。将富含十二烷的相导入含有 Novozyme 435 的酶反应器中，将脂肪醇和脂肪酸酯化成蜡酯。由于级联生物催化过程的成功简化，实现了蜡酯的连续生产（6.38–23.35 mg/(L·h)）。