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Production of Rebaudioside A from Stevioside Catalyzed by the Engineered Saccharomyces cerevisiae

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Abstract

Rebaudioside A has superior taste quality among the steviol glycosides extracted from Stevia rebaudiana leaves. Given its high purity as a general-purpose sweetener, rebaudioside A has received significant attention and has been widely applied in food and beverages in recent decades. Stevioside is one of the major steviol glycosides and can be converted to rebaudioside A by the uridine-diphosphate dependent glucosyltransferase UGT76G1 in S. rebaudiana. To explore the applicability of and limits in producing rebaudioside A from stevioside through whole-cell biocatalysis, the engineered Saccharomyces cerevisiae expressing UGT76G1, using a newly constructed constitutive expression vector, was used as the whole-cell biocatalyst. Citrate was added to the reaction mixture to allow metabolic regulation when glucose was fed to provide the activated sugar donor UDP-glucose for glycosylation of stevioside in vivo. In an evaluation of the whole-cell reaction parameters involving cell permeability, temperature, pH, citrate and Mg2+ concentrations, and glucose feeding, production of 1160.5 mg/L rebaudioside A from 2 g/L stevioside was achieved after 48 h without supplementation of extracellular UDP-glucose.

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Authors and Affiliations

  1. College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, Jiangsu, 211816, China

    Yan Li, Yangyang Li, Yu Wang, Liangliang Chen, Ming Yan, Kequan Chen, Lin Xu & Pingkai Ouyang

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  1. Yan Li
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  2. Yangyang Li
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  3. Yu Wang
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  7. Lin Xu
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  8. Pingkai Ouyang
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Correspondence to Yan Li.

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Funding

This work was financially supported by PAPD, NSFC (grant number 21106068), and Doctoral Fund of Ministry of Education of China (grant number 20113221120002).

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The authors declare that they have no competing interests.

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Li, Y., Li, Y., Wang, Y. et al. Production of Rebaudioside A from Stevioside Catalyzed by the Engineered Saccharomyces cerevisiae . Appl Biochem Biotechnol 178, 1586–1598 (2016). https://doi.org/10.1007/s12010-015-1969-4

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  • DOI: https://doi.org/10.1007/s12010-015-1969-4

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