Microbial lipids have the potential to displace terrestrial oils for fuel, value chemical, and food production, curbing the growth in tropical oil plantations and helping to reduce deforestation. However, commercialization remains elusive partly due to the lack of suitably robust organisms and their low lipid productivity. Extremely high cell densities in oleaginous cultures are needed to increase reaction rates, reduce reactor volume, and facilitate downstream processing. In this investigation, the oleaginous yeast Metschnikowia pulcherrima, a known antimicrobial producer, was cultured using four different processing strategies to achieve high cell densities and gain suitable lipid productivity. In batch mode, the yeast demonstrated lipid contents more than 40% (w/w) under high osmotic pressure. In fed-batch mode, however, high-lipid titers were prevented through inhibition above 70.0 g L −1 yeast biomass. Highly promising were a semi-continuous and continuous mode with cell recycle where cell densities of up to 122.6 g L −1 and maximum lipid production rates of 0.37 g L −1 h −1 (daily average), a nearly two-fold increase from the batch, were achieved. The findings demonstrate the importance of considering multiple fermentation modes to achieve high-density oleaginous yeast cultures generally and indicate the limitations of processing these organisms under the extreme conditions necessary for economic lipid production. K E Y W O R D Scontinuous, fed-batch, high cell density, microbial lipids, oleaginous yeast, semi-continuous

Macroalgae (seaweeds) represent an emerging resource for food and the production of commodity and specialty chemicals. In this study, a single-step microwave process was used to depolymerise a range of macroalgae native to the United Kingdom, producing a growth medium suitable for microbial fermentation. The medium contained a range of mono-and polysaccharides as well as macro-and micronutrients that could be metabolised by the oleaginous yeast Metschnikowia pulcherrima. Among twelve macroalgae species, the brown seaweeds exhibited the highest fermentation potential, especially the kelp Saccharina latissima. Applying a portfolio of ten native M. pulcherrima strains, yeast growth kinetics, as well as production of lipids and 2-phenylethanol were examined, with productivity and growth rate being strain dependent. On the 2 L scale, 6.9 g L −1 yeast biomassa yield of 0.14 g g −1 with respect to the supplied macroalgaecontaining 37.2% (w/w) lipid was achieved through utilisation of the proteins, mono-and polysaccharides from S. latissima, with no additional enzymes. In addition, the yeast degraded a range of fermentation inhibitors released upon microwave processing at high temperatures and long holding times. As macroalgae can be cultured to food grade, this system offers a novel, potentially low-cost route to edible microbial oils as well as a renewable feedstock for oleochemicals.

Lipid-based biofuels, such as biodiesel and hydroprocessed esters, are a central part of the global initiative to reduce the environmental impact of the transport sector. The vast majority of production is currently from first-generation feedstocks, such as rapeseed oil, and waste cooking oils. However, the increased exploitation of soybean oil and palm oil has led to vast deforestation, smog emissions and heavily impacted on biodiversity in tropical regions. One promising alternative, potentially capable of meeting future demand sustainably, are oleaginous yeasts. Despite being known about for 143 years, there has been an increasing effort in the last decade to develop a viable industrial system, with currently around 100 research papers published annually. In the academic literature, approximately 160 native yeasts have been reported to produce over 20% of their dry weight in a glyceride-rich oil. The most intensively studied oleaginous yeast have been Cutaneotrichosporon oleaginosus (20% of publications), Rhodotorula toruloides (19%) and Yarrowia lipolytica (19%). Oleaginous yeasts have been primarily grown on single saccharides (60%), hydrolysates (26%) or glycerol (19%), and mainly on the mL scale (66%). Process development and genetic modification (7%) have been applied to alter yeast performance and the lipids, towards the production of biofuels (77%), food/supplements (24%), oleochemicals (19%) or animal feed (3%). Despite over a century of research and the recent application of advanced genetic engineering techniques, the industrial production of an economically viable commodity oil substitute remains elusive. This is mainly due to the estimated high production cost, however, over the course of the twenty-first century where climate change will drastically change global food supply networks and direct governmental action will likely be levied at more destructive crops, yeast lipids offer a flexible platform for localised, sustainable lipid production. Based on data from the large majority of oleaginous yeast academic publications, this review is a guide through the history of oleaginous yeast research, an assessment of the best growth and lipid production achieved to date, the various strategies employed towards industrial production and importantly, a critical discussion about what needs to be built on this huge body of work to make producing a yeast-derived, more sustainable, glyceride oil a commercial reality.

BACKGROUND: Microbial oils -often termed single cell oils (SCOs) -offer an alternative to terrestrial oil crops across the energy, food and chemical industries. In addition to oils, a range of secondary metabolites can be produced from the heterotrophic organisms as part of a biorefinery system. Techno-economic analysis (TEA) is an important tool for evaluating economic viability, and although TEA is subject to high uncertainties where production is still at the laboratory scale, the tool can play a significant role in directing further research to evaluate suitability of scale-up.RESULTS: SCO production from the oleaginous yeast Metschnikowia pulcherrima using sucrose, wheat straw and distillery waste feedstocks was evaluated at two production scales. At a scale of 100 t year −1 oil production a minimum estimated selling price (MESP) of €14 000 t -1 was determined for sucrose. This reduced to €4-8000 t -1 on scaling to 10 000 t year −1 , with sucrose and wheat straw yielding the lowest MESP. CONCLUSIONS: Feedstock price and lipid yield had the greatest impact on overall economic return, though the valorization of co-products also had a large effect, and further play between feedstock and system productivity strategies could bring the price down to be competitive with terrestrial oils in the future. The novel approach demonstrated here for the first time integrates uncertainty into economic analysis whilst facilitating decision-support at an early technology development stage.(2) Discounted cash flow analysis was used to obtain a minimum estimated selling price (MESP) based on a net present value (NPV) J Chem Technol Biotechnol 2019; 94: 701-711

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