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Abstract
Mutagenesis and adaptation of the psychrotrophic fungus Chrysosporium pannorum A-1 to the toxic substrate β-pinene were used to obtain a biocatalyst with increased resistance to this terpene and improved bioconversion properties. Mutants of the parental strain were induced with UV light and N-methyl-N′-nitro-N-nitrosoguanidine. Mutants resistant to β-pinene were isolated using agar plates with a linear gradient of substrate concentrations. Active mutants were selected based on their general metabolic activity (GMA) expressed as oxygen consumption rate. Compared to the parental strain, the most active mutant showed an enhanced biotransformation ability to convert β-pinene to trans-pinocarveol (315 mg per g of dry mycelium), a 4.3-fold greater biocatalytic activity, and a higher resistance to H2O2-induced oxidative stress. Biotransformation using adapted mutants yielded twice as much trans-pinocarveol as the reaction catalyzed by non-adapted mutants. The results indicate that mutagenesis and adaptation of C. pannorum A-1 is an effective method of enhancing β-bioconversion of terpenes.
Highlights
Terpenes are valuable natural substrates commonly used in the production of fine chemicals.Turpentine, obtained from biomass and as a side product of the softwood industry, is rich in monoterpenes, such as α-pinene and β-pinene, which are widely used as raw materials in the synthesis of flavors, fragrances, and pharmaceutical compounds [1,2]
We demonstrated that the psychrotrophic fungus Chrysosporium pannorum
It has been shown that the effectiveness of biotransformation largely relies on the interactions between the biocatalyst and environmental stressors, which may increase the yields of biotransformation products [44]
Summary
Terpenes are valuable natural substrates commonly used in the production of fine chemicals.Turpentine, obtained from biomass and as a side product of the softwood industry, is rich in monoterpenes, such as α-pinene and β-pinene, which are widely used as raw materials in the synthesis of flavors, fragrances, and pharmaceutical compounds [1,2]. Many of them have beneficial effects on health [3,4,5], including anti-inflammatory activity [6] Their high volatility, poor solubility in aqueous media, and natural toxicity to microorganisms still limit the use of these compounds in applied biocatalysis. These limitations can be overcome by slow/gradual addition of terpene into the reaction medium, use of biphasic systems containing biocompatible organic solvents or a co-solvent, and in situ product removal techniques [7,8,9]
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