Abstract
The biocatalytic conversion of sclareol to ambradiol, a valuable component in the fragrance industry, using whole-cell biotransformation by the dimorphic yeast Hyphozyma roseoniger, was investigated using metabolomics tools. An integrated approach was used to identify and quantify the participating intermediates in this bioconversion using both nuclear magnetic resonance (NMR) spectroscopy and liquid chromatography coupled to mass spectrometry (LC–MS). This study entailed growth stage-dependent analysis of H. roseoniger suspensions grown in batch culture over a 14-day period, beginning with a three-day induction period using 20 mg/200 mL sclareol, followed by a further 1 g/200 mL sclareol dose to enable ambradiol production. The progress of the bioconversion and the resulting dynamic changes to the metabolome were monitored using NMR analysis and semi-targeted LC–MS metabolomics. This outlined the molecular conversions occurring within the matrix and no novel intermediates participating in the sclareol to ambradiol conversion could be identified. This study presents new findings about the transformative capabilities of H. roseoniger as a whole cell biocatalyst, highlighting its potential utility in similar applications.
Highlights
By definition, biocatalysis involves the utilization of soluble or immobilized microbial enzymes, whole cell catalysts, de novo microbial processes such as fermentation, or cell extracts for specific bioconversion or biotransformation of organic precursors, leading to different functionalized pure compounds [1,2,3]
Using semi-targeted metabolomics tools, signature biomarkers at each stage of the optimized reaction outlined by Steenkamp and Taka [17] were targeted for quantitation
Based on its structure, it is uncertain if cis-abienol is part of the conversion sequence from sclareol to ambradiol or if it is indicative of an alternative metabolism event in the cells
Summary
Biocatalysis involves the utilization of soluble or immobilized microbial enzymes, whole cell catalysts, de novo microbial processes such as fermentation, or cell extracts for specific bioconversion or biotransformation of organic precursors, leading to different functionalized pure compounds [1,2,3]. Increased interest in using biocatalytic processes for production of desired chemicals is based on the perceived eco-friendliness of the method [2,4,5] low productivity may detract from its application. There is ongoing research on biocatalytic routes for ambrafuran and other ambergris-related odorants as alternative synthetic routes [8] Examples of these techniques include fermentation processes based on synthetic biology, white biotechnology (that involves the use of living cells and enzymes) to biosynthesize ambrafuran, by means of green chemistry technologies (environmentally benign chemical products and processes). These approaches are recommendable as they require less energy and create less waste [2,9]
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