| Summary: | Recently discovered endogenous mammalian lipids fatty acid esters of hydroxy fatty acids (FAHFAs), proved to have anti-inflammatory and anti-diabetic effects . Due to their extremely low abundancies in vivo , forging a feasible scenario for FAHFA synthesis is critical for their use in uncovering biological mechanism or clinical trials. Here, we showcase a fully enzymatic approach, a novel in vitro bi-enzymatic cascade system, enabling an effective conversion of nature-abundant fatty acid into FAHFAs. Two hydratases from L. acidophilus were used for converting unsaturated fatty acids to various stereospecific hydroxy fatty acids, followed by esterification with another fatty acid catalyzed by C. antarctica lipase A (CALA). Various FAHFAs were synthesized in a preparative scale using this bi-enzymatic approach in a one-pot two-step operation mode. In all, we demonstrated that hydratase-CALA system promises a sustainable solution to the synthesis of structure-diverse stereospecific FAHFAs.
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