Carbohydrate Spectrum of Extremophilic Yeasts Yarrowia lipolytica under pH Stress

Sekova, V. Yu.; Дергачева, Д. И.; Терешина, В. М.; Исакова, Е. П.; Deryabina, Yulia I.

doi:10.1134/s0026261718020133

Cited by 9 publications

(6 citation statements)

References 38 publications

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“…Although Y. lipolytica is one of the best erythritol producers, it also presents some drawbacks such as the synthesis of byproducts including mannitol and d -arabitol. This negatively impacts the yield from glucose and complexifies the purification steps [ 9 , 13 , 19 – 22 ]. Therefore, we thought of constructing a strain unable to produce these byproducts.…”

Section: Resultsmentioning

confidence: 99%

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

Wang

Chi

Zou

et al. 2020

Biotechnol Biofuels

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Background Functional sugar alcohols have been widely used in the food, medicine, and pharmaceutical industries for their unique properties. Among these, erythritol is a zero calories sweetener produced by the yeast Yarrowia lipolytica. However, in wild-type strains, erythritol is produced with low productivity and yield and only under high osmotic pressure together with other undesired polyols, such as mannitol or d-arabitol. The yeast is also able to catabolize erythritol in non-stressing conditions. Results Herein, Y. lipolytica has been metabolically engineered to increase erythritol production titer, yield, and productivity from glucose. This consisted of the disruption of anabolic pathways for mannitol and d-arabitol together with the erythritol catabolic pathway. Genes ZWF1 and GND encoding, respectively, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were also constitutively expressed in regenerating the NADPH2 consumed during erythritol synthesis. Finally, the gene RSP5 gene from Saccharomyces cerevisiae encoding ubiquitin ligase was overexpressed to improve cell thermoresistance. The resulting strain HCY118 is impaired in mannitol or d-arabitol production and erythritol consumption. It can grow well up to 35 °C and retain an efficient erythritol production capacity at 33 °C. The yield, production, and productivity reached 0.63 g/g, 190 g/L, and 1.97 g/L·h in 2-L flasks, and increased to 0.65 g/g, 196 g/L, and 2.51 g/L·h in 30-m3 fermentor, respectively, which has economical practical importance. Conclusion The strategy developed herein yielded an engineered Y. lipolytica strain with enhanced thermoresistance and NADPH supply, resulting in a higher ability to produce erythritol, but not mannitol or d-arabitol from glucose. This is of interest for process development since it will reduce the cost of bioreactor cooling and erythritol purification.

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Section: Resultsmentioning

confidence: 99%

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

Wang

Chi

Zou

et al. 2020

Biotechnol Biofuels

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“…Probably, both hypotheses are correct, and mannitol can be localized in a cell at the specific site of high HO • production or in the close vicinity of the key HO • target molecules [9]. Based on our previous studies [40], we could suppose that constitutively high concentrations of mannitol in the Y. lipolytica yeast facilitated its resistance to extreme conditions by effective scavenging of ROS generated at extreme pH. No significant difference in the mannitol concentration in the culture grown at different pH (Figure 2) suggests that the role of mannitol as a factor of pH adaptation should be excluded.…”

Section: Cytosol Carbohydrate Profilementioning

confidence: 95%

“…The accumulation of D-arabitol and trehalose was shown for the Debaryomyces and Geotrichum fungi [46], the Kluyveromyces lactis yeast [47], and the Candida albicans yeast [10] to the exposure of thermal, oxidative, and osmotic stress factors. In the eukaryotic cells, D-arabitol is known as a by-product of the pentose phosphate pathway in some processes, such as a supply of the pool of NADPH-reducing equivalents [40]. It is similar to mannitol, which can be oxidized to fructose, and is also capable of reducing NADP to NADPH [16].…”

Section: Cytosol Carbohydrate Profilementioning

confidence: 99%

Soluble Sugar and Lipid Readjustments in the Yarrowia lipolytica Yeast at Various Temperatures and pH

et al. 2019

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Microorganisms cope with a wide range of environmental challenges using different mechanisms. Their ability to prosper at extreme ambient pH and high temperatures has been well reported, but the adaptation mechanism often remains unrevealed. In this study, we addressed the dynamics of lipid and sugar profiles upon different cultivation conditions. The results showed that the cells grown at various pH and optimal temperature contained mannitol as the major cytosol sugar alcohol. The elevated temperature of 38 °C led to a two- to three-fold increase in total cytosol sugars with concurrent substitution of mannitol for trehalose. Lipid composition in the cells at optimal temperature changed insignificantly at any pH tested. The increase in the temperature caused some drop in the storage and membrane lipid levels, remarkable changes in their composition, and the degree of unsaturated fatty acids. It was shown that the fatty acid composition of some membrane phospholipids varied considerably at changing pH and temperature values. The data showed a pivotal role and flexibility of the sugar and lipid composition of Y. lipolytica W29 in adaptation to unfavorable environmental conditions.

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“…Mannitol production can alleviate the redox imbalance of yeast cells by either recycling NADP + when NADPH is in excess or providing the NADPH required during lipid production [25,26]. In addition, mannitol production has also been related to stress-protective mechanisms to reactive oxygen species (ROS) [27] and/ or to thermal stress [21]. In this work, depending on the cultivation media, maximum mannitol concentrations varied between 10 and 28 g/L.…”

Section: Pulse-feeding Cultivationmentioning

confidence: 95%

Efficient use of discarded vegetal residues as cost-effective feedstocks for microbial oil production

Gallego-García

Moreno

González

et al. 2023

Biotechnol Biofuels

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Background Horticultural intensive type systems dedicated in producing greenhouse vegetables are one of the primary industries generating organic waste. Towards the implementation of a zero-waste strategy, this work aims to use discarded vegetables (tomato, pepper and watermelon) as feedstock for producing microbial oil using the oleaginous yeast Cryptococcus curvatus. Results The soluble fraction, resulting after crushing and centrifuging these residues, showed C/N ratios of about 15, with a total carbohydrate content (mainly glucose, fructose and sucrose) ranging from 30 g/L to 65 g/L. Using these liquid fractions as substrate under a pulse-feeding strategy with a concentrated glucose solution resulted in an intracellular total lipid accumulation of about 30% (w/w) of the total dry cell weight (DCW). To increase this intracellular lipid content, the initial C/N content was increased from 15 to 30 and 50. Under these conditions, the process performance of the pulse-feeding strategy increased by 20–36%, resulting in a total intracellular lipid concentration of 35–40% DCW (w/w). Conclusion These results demonstrate the potential of discarded vegetables as a substrate for producing bio-based products such as microbial oil when proper cultivation strategies are available.

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Carbohydrate Spectrum of Extremophilic Yeasts Yarrowia lipolytica under pH Stress

Cited by 9 publications

References 38 publications

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity

Soluble Sugar and Lipid Readjustments in the Yarrowia lipolytica Yeast at Various Temperatures and pH

Efficient use of discarded vegetal residues as cost-effective feedstocks for microbial oil production

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