Abstract

BackgroundHeterotrophic microbial oils are potentially a more sustainable alternative to vegetable or fossil oils for food and fuel applications. However, as almost all work in the area is conducted on the laboratory scale, such studies carry limited industrial relevance and do not give a clear indication of what is required to produce an actual industrial process. Metschnikowia pulcherrima is a non-pathogenic industrially promising oleaginous yeast which exhibits numerous advantages for cost-effective lipid production, including a wide substrate uptake, antimicrobial activity and fermentation inhibitor tolerance. In this study, M. pulcherrima was fermented in stirred tank reactors of up to 350 L with 250-L working volume in both batch and semi-continuous operation to highlight the potential industrial relevance. Due to being food-grade, suitable for handling at scale and to demonstrate the oligosaccharide uptake capacity of M. pulcherrima, enzyme-hydrolysed starch in the form of glucose syrup was selected as fermentation feedstock.ResultsIn batch fermentations on the 2-L scale, a lipid concentration of 14.6 g L−1 and productivity of 0.11 g L−1 h−1 were achieved, which was confirmed at 50 L (15.8 g L−1; 0.10 g L−1 h−1). The maximum lipid production rate was 0.33 g L−1 h−1 (daily average), but the substrate uptake rate decreased with oligosaccharide chain length. To produce 1 kg of dry yeast biomass containing up to 43% (w/w) lipids, 5.2 kg of the glucose syrup was required, with a lipid yield of up to 0.21 g g−1 consumed saccharides. In semi-continuous operation, for the first time, an oleaginous yeast was cultured for over 2 months with a relatively stable lipid production rate (around 0.08 g L−1 h−1) and fatty acid profile (degree of fatty acid saturation around 27.6% w/w), and without contamination. On the 250-L scale, comparable results were observed, culminating in the generation of nearly 10 kg lipids with a lipid productivity of 0.10 g L−1 h−1.ConclusionsThe results establish the importance of M. pulcherrima for industrial biotechnology and its suitability to commercially produce a food-grade oil. Further improvements in the productivity are required to make M. pulcherrima lipid production industrial reality, particularly when longer-chain saccharides are involved.

Highlights

  • Heterotrophic microbial oils are potentially a more sustainable alternative to vegetable or fossil oils for food and fuel applications

  • Hydrolysed starch was shown to be a suitable feedstock for M. pulcherrima, with the lipid yields with respect to the consumed saccharides considerably exceeding those using any other feedstock to date, including glucose and glycerol

  • The results demonstrate that M. pulcherrima is an oleaginous yeast suitable for continued operation, with a reasonably steady lipid production rate, fatty acid composition of the produced oil and no contamination in fermentation over 2 months

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Summary

Introduction

Heterotrophic microbial oils are potentially a more sustainable alternative to vegetable or fossil oils for food and fuel applications. Metschnikowia pulcherrima is a non-pathogenic industrially promising oleaginous yeast which exhibits numerous advantages for cost-effective lipid production, including a wide substrate uptake, antimicrobial activity and fermentation inhibitor tolerance. Due to being food-grade, suitable for handling at scale and to demonstrate the oligosaccharide uptake capacity of M. pulcherrima, enzyme-hydrolysed starch in the form of glucose syrup was selected as fermentation feedstock. It has been shown that Metschnikowia pulcherrima is a very promising oleaginous yeast for commercial lipid production, growing on a variety of low-cost substrates in non-sterile environments [8, 16]. As being foodgrade, hydrolysed starch in the form of glucose syrup (GS) is potentially a suitable feedstock for the production of a food-grade microbial oil, though the economic feasibility would depend heavily on the characteristics and price of the produced oil and by-products [13, 14]. Considering M. pulcherrima’s antagonistic traits facilitating sterility [22, 23] and capability of producing an oil similar in composition to prominent vegetable oils including palm oil [3, 16], the development of a foodgrade oil can be envisioned in addition to biodiesel

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