Bacterial Whole Cells Synthesis of Whisky Lactones in a Solid-State Fermentation Bioreactor Prototype

Agro-industrial side streams such as oilseed cakes were used as a medium in solid-state fermentation (SSF) for microbial oxidation of anti- and syn-3-methyl-octane-1,4-diols to obtain corresponding trans- and cis-whisky lactones. In preliminary screening transformations, a wide range of whole bacterial cells were tested on the basis of oxidation activity, which is rarely described in the literature, in contrast to the widely studied lipolytic activity on SSF. Among the different oil cakes tested, biotransformations carried out on linseed cake were characterized by the highest conversion and stereoselectivity. Several preparative-scale oxidations performed in a self-constructed SSF bioreactor catalyzed by Rhodococcus erythropolis DSM44534, Rhodococcus erythropolis PCM2150 and Gordonia rubripertincta PCM2144 afforded optically active trans-(+)-(4S,5R), cis-(+)-(4R,5R) and cis-(-)-(4S,5S) isomers of whisky lactones, respectively. Bacteria of the Rhodococcus, Gordonia, Dietzia and Streptomyces genera carried out transformations with complete conversion after three days. Various extraction methods were applied for the isolation of the products, and among them, the combination of steam distillation with simple extraction were the most efficient. Biotransformations were conducted under precise control of conditions in a bioreactor based on a Raspberry Pi Zero W. The proposed low-cost (ca. USD 100) bioreactor is a standalone system that is fully autoclavable and easy to use.