Abstract

The psychrotrophic fungus Chrysosporium pannorum A-1 is reported for the first time as a novel biocatalyst for O2-promoted oxidation of α-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 α-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 α-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 °C, 10 °C being the optimum for the production of 1 and 20 °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 α-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 α-pinene by a wild-type microorganism.Electronic supplementary materialThe online version of this article (doi:10.1007/s10295-014-1550-0) contains supplementary material, which is available to authorized users.

Highlights

  • Cold-adapted microorganisms have the ability to reproduce and grow at temperatures close to 0 °C

  • The fungus Ch. pannorum A-1 was chosen for the study from among a group of psychrotrophic microorganisms showing good activity in the biotransformation of R-(+)limonene [45]

  • The dynamics of mycelial growth in the liquid medium at 20 °C, as graphically depicted in Fig. 1, reveal the three phases normally found in physiological characteristics and show that the fungus grew at a high rate, reaching the stationary phase already after 60 h of growth

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Summary

Introduction

Cold-adapted (psychrophilic and psychrotrophic) microorganisms have the ability to reproduce and grow at temperatures close to 0 °C. Organisms native to cold environments achieve metabolic rates that are only slightly lower than those reached by organisms living at moderate temperatures. Due to their high enzymatic activities and catalytic efficiencies in the temperature range of 0–20 °C, cold-adapted microorganisms are useful for biotransformation involving volatile compounds, such as flavors and fragrances subject to faster evaporation at higher temperatures making it possible to obtain high amounts of these valuable products. A microbial reaction system based on psychrotrophic Mortierella minutissima, which shows a high catalase activity, has been developed for the oxidation of this monoterpene,

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