A scalable whole-cell biocatalytic process for food-compatible production of resolvin D5 using the edible mushroom Pleurotus ostreatus.

Abstract

Specialized pro-resolving mediators (SPMs) actively promote the resolution of inflammation, but their scalable and food-compatible production remains challenging because current methods rely on multistep chemical synthesis, mammalian cells, purified enzymes, crude enzyme preparations, or recombinant microorganisms. Here, we developed a scalable, food-compatible submerged-cultured whole-cell biocatalytic process for the production of resolvin D5 (RvD5; 7S,17S-dihydroxydocosahexaenoic acid) from docosahexaenoic acid (DHA) using the edible mushroom Pleurotus ostreatus. Submerged-cultured P. ostreatus whole cells stereoselectively converted DHA to RvD5 via a 17S-hydroxydocosahexaenoic acid intermediate through an endogenous lipoxygenase (LOX)-mediated double-oxygenation pathway, without enzyme purification or heterologous gene expression. Optimization of submerged cultivation conditions (cultivation period and medium composition) increased RvD5 production by 2.1-fold, and subsequent optimization of whole-cell biotransformation parameters (temperature, pH, cell and substrate concentrations, polyvinylpyrrolidone and ethanol supplementation, reaction time, and agitation) resulted in an overall 13.4-fold increase relative to the initial conditions. Under the optimized flask-scale conditions, whole cells produced 2.14mM (0.77g/L) RvD5 from 2.5mM DHA with an 85.6% molar yield within 40min. The process was successfully scaled up to a 3.0-L stirred-tank bioreactor (1.0-L working volume), producing 2.08mM (0.75g/L) RvD5 with an 83.2% molar yield within 60min. The developed cultivation-based edible fungal whole-cell biocatalytic process integrates catalyst preparation, efficient RvD5 biosynthesis, and scalable bioprocessing, providing a practical strategy for the scalable biotechnological production of DHA-derived SPMs.

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