Oczyszczanie i charakterystyka dehydrogenazy erytrytolowej z &lt;i&gt;Mycobacterium smegmatis&lt;/i&gt;

Paradowska, Katarzyna; Nitka, Dagmara

doi:10.2478/v10080-008-0177-8

Cited by 4 publications

(3 citation statements)

References 9 publications

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“…DHAP: dihydroxyacetone phosphate; GA3P: glyceraldehyde-3-phosphate; GK: glycerol kinase; GDH: glycerol-3P dehydrogenase; TIM: triosephosphate isomerase; TK: transketolase; E4PP: erythrose-4P phosphatase; ER: erythrose reductase; and EK: erythrulose kinase. Dotted arrows represent the hypothetic catabolism pathway of erythrulose-P according to Paradowska and Nitka (2009). designed to introduce an AvrII site at the 3′ end and either a BamHI or a BglII restriction site at the 5′ end (Supplementary table 2).…”

Section: Strain Constructionmentioning

confidence: 99%

Enhancing erythritol productivity in Yarrowia lipolytica using metabolic engineering

Carly

Vandermies

Telek

et al. 2017

Metabolic Engineering

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Erythritol (1,2,3,4-butanetetrol) is a four-carbon sugar alcohol with sweetening properties that is used by the agrofood industry as a food additive. In this study, we demonstrated that metabolic engineering can be used to improve the production of erythritol from glycerol in the yeast Yarrowia lipolytica. The best results were obtained using a mutant that overexpressed GUT1 and TKL1, which encode a glycerol kinase and a transketolase, respectively, and in which EYK1, which encodes erythrulose kinase, was disrupted; the latter enzyme is involved in an early step of erythritol catabolism. In this strain, erythritol productivity was 75% higher than in the wild type; furthermore, the culturing time needed to achieve maximum concentration was reduced by 40%. An additional advantage is that the strain was unable to consume the erythritol it had created, further increasing the process's efficiency. The erythritol productivity values we obtained here are among the highest reported thus far.

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Section: Strain Constructionmentioning

confidence: 99%

Enhancing erythritol productivity in Yarrowia lipolytica using metabolic engineering

Carly

Vandermies

Telek

et al. 2017

Metabolic Engineering

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“…In a first one, erythritol is phosphorylated into erythritol-phosphate and then oxidized in erythrulose-phosphate [ 44 ]. In a second one, erythritol is first converted into erythrulose before being phosphorylated into erythrulose-phosphate [ 45 ]. We have recently identified and characterized EYK1 gene (YALI0F1606g) [ 46 ] in Y. lipolytica .…”

Section: Resultsmentioning

confidence: 99%

“…1 Erythritol catabolism pathway in Yarrowia lipolytica . Based on [ 45 ] and [ 46 ]. Erythritol is converted into l -erythrulose by an erythritol dehydrogenase encoded by EYD1 , then l -erythrulose is phosphorylated by the l -erythrulose-1-kinase encoded by EYK1 (YALI0F01606g) into l -erythrulose-1-phosphate …”

Section: Resultsmentioning

confidence: 99%

New inducible promoter for gene expression and synthetic biology in Yarrowia lipolytica

et al. 2017

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BackgroundThe oleaginous yeast Yarrowia lipolytica is increasingly used as alternative cell factory for the production of recombinant proteins. At present, several promoters with different strengths have been developed based either on the constitutive pTEF promoter or on oleic acid inducible promoters such as pPOX2 and pLIP2. Although these promoters are highly efficient, there is still a lack of versatile inducible promoters for gene expression in Y. lipolytica. ResultsWe have isolated and characterized the promoter of the EYK1 gene coding for an erythrulose kinase. pEYK1 induction was found to be impaired in media supplemented with glucose and glycerol, while the presence of erythritol and erythrulose strongly increased the promoter induction level. Promoter characterization and mutagenesis allowed the identification of the upstream activating sequence UAS1EYK1. New hybrid promoters containing tandem repeats of either UAS1XPR2 or UAS1EYK1 were developed showing higher expression levels than the native pEYK1 promoter. Furthermore, promoter strength was improved in a strain carrying a deletion in the EYK1 gene, allowing thus the utilization of erythritol and erythrulose as free inducer.ConclusionsNovel tunable and regulated promoters with applications in the field of heterologous protein production, metabolic engineering, and synthetic biology have been developed, thus filling the gap of the absence of versatile inducible promoter in the yeast Y. lipolytica.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-017-0755-0) contains supplementary material, which is available to authorized users.

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Identification and characterization of EYK1, a key gene for erythritol catabolism in Yarrowia lipolytica

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Gamboa-Meléndez

Vandermies

et al. 2017

Appl Microbiol Biotechnol

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Erythritol is a four-carbon sugar alcohol synthesized by osmophilic yeasts, such as Yarrowia lipolytica, in response to osmotic stress. This metabolite has application as food additive due to its sweetening properties. Although Y. lipolytica can produce erythritol at a high level from glycerol, it is also able to consume it as carbon source. This ability negatively affects erythritol productivity and represents a serious drawback for the development of an efficient erythritol production process. In this study, we have isolated by insertion mutagenesis a Y. lipolytica mutant unable to grow on erythritol. Genomic characterization of the latter highlighted that the mutant phenotype is directly related to the disruption of the YALI0F01606g gene. Several experimental evidences suggested that the identified gene, renamed EYK1, encodes an erythrulose kinase. The mutant strain showed an enhanced capacity to produce erythritol as compared to the wild-type strain. Moreover, in specific experimental conditions, it is also able to convert erythritol to erythrulose, another compound of biotechnological interest.

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Oczyszczanie i charakterystyka dehydrogenazy erytrytolowej z <i>Mycobacterium smegmatis</i>

Cited by 4 publications

References 9 publications

Enhancing erythritol productivity in Yarrowia lipolytica using metabolic engineering

Enhancing erythritol productivity in Yarrowia lipolytica using metabolic engineering

New inducible promoter for gene expression and synthetic biology in Yarrowia lipolytica

Identification and characterization of EYK1, a key gene for erythritol catabolism in Yarrowia lipolytica

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