The mitochondrial citrate carrier in Yarrowia lipolytica: Its identification, characterization and functional significance for the production of citric acid

Yuzbasheva, Evgeniya Y.; Agrimi, Gennaro; Yuzbashev, Tigran V.; Scarcia, Pasquale; Vinogradova, E. B.; Palmieri, Luigi; Shutov, Artem V.; Kosikhina, Iuliia M.; Palmieri, Ferdinando; Sineoky, S. P.

doi:10.1016/j.ymben.2019.05.002

Cited by 60 publications

(47 citation statements)

References 70 publications

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“…The C4 dicarboxylates fumarate and oxaloacetate were also transported at high rate by both proteins and, at least in principle, could be considered alternative candidate countersubstrates of cis‐aconitate. Interestingly, the substrate specificity of At_MttA overlaps that of the recently studied Y. lipolytica mitochondrial carrier YlYhm2 , involved in the citrate production and lipogenesis, with the only exception of a swap of the transported tricarboxylate: cis‐aconitate in place of citrate. It should be emphasized that even if Um_Mtt1 and At_MttA cluster with the tricarboxylate mitochondrial carriers, they are not able to transport citrate, which would be detrimental to IA biosynthesis.…”

Section: Discussionmentioning

confidence: 88%

“…Overexpression of the recombinant proteins Um_Mtt1 and At_MttA was carried out at 37 °C in the E. coli strains Rosetta(DE3) (Novagen, Darmstadt, Germany) and M15(pREP4) (QIAGEN Inc.), respectively, as previously described . Control cultures with the empty vectors were processed in parallel.…”

Section: Methodsmentioning

confidence: 99%

“…Control cultures with the empty vectors were processed in parallel. Inclusion bodies were purified from cellular content on sucrose density gradient and washed at 4 °C once in TNS 3% buffer (Triton X‐114 3% w/v, 1 m m EDTA, 10 m m PIPES pH 7.4) and twice in TE buffer (10 m m Tris/HCl pH 7.4, 1 m m EDTA pH 7.4) . The identity of the two expressed proteins was verified by mass spectrometry as reported before .…”

Section: Methodsmentioning

confidence: 99%

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Mitochondrial carriers of Ustilago maydis and Aspergillus terreus involved in itaconate production: same physiological role but different biochemical features

et al. 2019

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Itaconic acid (IA) is a naturally occurring dicarboxylic acid with applications in the manufacture of polymers. IA can be produced by fermentation using the fungi Aspergillus terreus or Ustilago maydis as biocatalysts. Indirect evidence has suggested that the mitochondrial carriers U. maydis Um_Mtt1 and A. terreus At_MttA export mitochondrially synthesized cis‐aconitate to the cytosol for IA synthesis using malate as a countersubstrate. Here, by assaying the transport features of recombinant Um_Mtt1 and At_MttA in reconstituted liposomes, we find that both proteins efficiently transport cis‐aconitate, but malate is well transported only by Um_Mtt1 and 2‐oxoglutarate only by At_MttA. Bioinformatic analysis shows that Um_Mtt1 and At_MttA form a distinctive mitochondrial carrier subfamily. Our data show that although fulfilling the same physiological task, Um_Mtt1 and At_MttA have different biochemical features.

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Section: Discussionmentioning

confidence: 88%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Mitochondrial carriers of Ustilago maydis and Aspergillus terreus involved in itaconate production: same physiological role but different biochemical features

et al. 2019

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“…This results in a disrupted Krebs cycle with accumulation of early metabolites in the mitochondria, such as citric acid [13,14]. Citric acid can be finally exported from the mitochondria by exchange of cytosolic oxaloacetate via the specific transporter Yhm2p to accumulate extra-cellularly [15]. Therefore, at high C/N ratio, the excess of carbon is redirected toward CA synthesis during the stationary phase.…”

Section: Citric Acidmentioning

confidence: 99%

“…Pyruvate carboxylase (YALI0C24101g, PYC) catalyzes the carboxylation of pyruvate into oxaloacetate in the cytosol in the presence of CaCO 3 used as a source of carbon dioxide [45]. Expression of pyruvate carboxylase from Meyerozyma guiliermondii in Y. lipolytica SWJ-1b resulted in an increased CA titer of 101 g/L (with a resultant yield from glucose 0.89 g/g) within 240 h. Co-expression of YHM2 (YALI0B10736g) and AMDP encoding, respectively, mitochondrial citrate carrier and adenosine monophosphate deaminase in strain W29 allowed a CA titer and yield from glucose of 97.1 g/L and 0.5 g/g during culture in bioreactor [15]. In those conditions, the CA productivity was 0.93 g/(L•h).…”

Section: Citric Acidmentioning

confidence: 99%

Sugar Alcohols and Organic Acids Synthesis in Yarrowia lipolytica: Where Are We?

2020

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Sugar alcohols and organic acids that derive from the metabolism of certain microorganisms have a panoply of applications in agro-food, chemical and pharmaceutical industries. The main challenge in their production is to reach a productivity threshold that allow the process to be profitable. This relies on the construction of efficient cell factories by metabolic engineering and on the development of low-cost production processes by using industrial wastes or cheap and widely available raw materials as feedstock. The non-conventional yeast Yarrowia lipolytica has emerged recently as a potential producer of such metabolites owing its low nutritive requirements, its ability to grow at high cell densities in a bioreactor and ease of genome edition. This review will focus on current knowledge on the synthesis of the most important sugar alcohols and organic acids in Y. lipolytica.

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A robust soft sensor based on artificial neural network for monitoring microbial lipid fermentation processes using Yarrowia lipolytica

Wang

Zhao

Lin

et al. 2022

Biotech & Bioengineering

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Microbial oils produced by Yarrowia lipolytica offer an environmentally friendly and sustainable alternative to petroleum as well as traditional lipids from animals and plants. The accurate measurement of fermentation parameters, including the substrate concentration, dry cell weight, and lipid accumulation, is the foundation of process control, which is indispensable for industrial lipid production. However, it remains a great challenge to measure the complex parameters online during the lipid fermentation process, which is nonlinear, multivariate, and characterized by strong coupling. As a type of AI technology, the artificial neural network model is a powerful tool for handling extremely complex problems, and it can be employed to develop a soft sensor to monitor the microbial lipid fermentation process of Y. lipolytica. In this study, we first analyzed and emphasized the volume of sodium hydroxide and dissolved oxygen concentration as central parameters of the fermentation process. Then, a soft sensor based on a four-input artificial neural network model was developed, in which the input variables were fermentation time, dissolved oxygen concentration, initial glucose concentration, and additional volume of sodium hydroxide. This provides the possibility of online monitoring of dry cell weight, glucose concentration, and lipid production with high accuracy, which can be extended to similar fermentation processes characterized by the addition of bases or acids, as well as changes of the dissolved oxygen concentration.

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The mitochondrial citrate carrier in Yarrowia lipolytica: Its identification, characterization and functional significance for the production of citric acid

Cited by 60 publications

References 70 publications

Mitochondrial carriers of Ustilago maydis and Aspergillus terreus involved in itaconate production: same physiological role but different biochemical features

Mitochondrial carriers of Ustilago maydis and Aspergillus terreus involved in itaconate production: same physiological role but different biochemical features

Sugar Alcohols and Organic Acids Synthesis in Yarrowia lipolytica: Where Are We?

A robust soft sensor based on artificial neural network for monitoring microbial lipid fermentation processes using Yarrowia lipolytica

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