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Production of Arabidopsis thaliana UDP-Sugar Pyrophosphorylase by Pichia pastoris and Its Application in Efficient UDP-Glucose and UDP-Glucuronic Acid Synthesis

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Abstract

Sugar nucleotides, the activated forms of monosaccharides, are used as donor substrates by glycosyltransferases in the in vivo biosynthesis of polysaccharides, proteoglycans and glycoglycerolipids. In mammals, uridine 5'-diphosphate-glucose (UDP-Glc) and uridine 5'-diphosphate-glucuronic acid (UDP-GlcA) are common building blocks of glycans and glycoconjugates. The commercial demand for these high-energy donors is increasing, while the understanding and application of enzymatic glycan synthesis has leapt forward in recent years. To produce valuable UDP-GlcA in a cost-effective way, here, UDP-sugar pyrophosphorylase from Arabidopsis thaliana was constitutively expressed in Pichia pastoris and secreted into the extracellular medium. Under these conditions, the synthesis of 4.2 g UDP-GlcA or 5.5 g UDP-Glc per liter of culture was revealed in the culture medium, without any need for purification. An anion exchange chromatography purification method for UDP-sugars was also developed. This route opens a door to large-scale production of the cheaper UDP-GlcA.

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Funding

This work was supported by grants from the National Natural Science Foundation of China (project no. 31770845) and the Qilu Scholar Program of Shandong University.

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    Authors and Affiliations

    1. Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), School of Pharmaceutical Sciences, Shandong University, 250012, Jinan, China

      L. X. Zang, R. R. Du, H. C. Zang, F. S. Wang & J. Z. Sheng

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    1. L. X. Zang
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    2. R. R. Du
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    3. H. C. Zang
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    4. F. S. Wang
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    5. J. Z. Sheng
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    Correspondence to F. S. Wang or J. Z. Sheng.

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    The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

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    Zang, L.X., Du, R.R., Zang, H.C. et al. Production of Arabidopsis thaliana UDP-Sugar Pyrophosphorylase by Pichia pastoris and Its Application in Efficient UDP-Glucose and UDP-Glucuronic Acid Synthesis. Appl Biochem Microbiol 55, 631–638 (2019). https://doi.org/10.1134/S0003683819060152

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