Bioconversion of α-pinene by a novel cold-adapted fungus Chrysosporium pannorum

The psychrotrophic fungus Chrysosporium pannorum A-1 is reported for the first time as a novel biocatalyst for O-2-promoted oxidation of alpha-pinene. GC-MS analysis indicated that the main products of the reaction were compounds of a high commercial value, verbenol (1) and verbenone (2). Exponentially growing cells (days 2-3) were about twice as active as cells in the late stationary phase in terms of the total concentration of products. The highest yields of 1 and 2 were obtained using three-day and two-day-old mycelia and a medium containing 1.5 and 1 % (v/v) of the substrate, respectively. The optimal time for the bioconversion of alpha-pinene varied from 1 to 3 days, and depended on the kind of product desired. Most of 1 was produced at a relatively high concentration of 360 mg/L after the first six hours of alpha-pinene bioconversion [with an average yield of 69 mg/(g (dry cell) L aqueous phase)]. The oxidative activity of C. pannorum was identified across a wide temperature range of 5-25 A degrees C, 10 A degrees C being the optimum for the production of 1 and 20 A degrees C for the production of 2. Sequential addition of the substrate during 3 days of the biotransformation resulted in a significant increase in 1 and 2 up to 722 and 176 mg/L, respectively, and a 2-fold enhancement of product yield as compared to bioconversion with a single supply of alpha-pinene. The concentration of total conversion products in the culture medium reached 1.33 g/L [which corresponded product yield of 225 mg/(g (dry cell) L)]. This represents probably the most promising result reported to date for oxidative biotransformation of alpha-pinene by a wild-type microorganism.