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Pilot studies on scale-up biocatalysis of 7-β-xylosyl-10-deacetyltaxol and its analogues by an engineered yeast

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Paclitaxel content in yew tree is extremely low, causing a worldwide shortage of this important anticancer drug. Yew tree can also produce abundant 7-β-xylosyl-10-deacetyltaxol that can be bio-converted into 10-deacetyltaxol for semi-synthesis of paclitaxel. However, the bio-conversion by the screened natural microorganisms was inefficient. We have constructed the recombinant yeast with a glycoside hydrolase gene from Lentinula edodes and explored the bioconversion. Based on previously established reaction conditions, the bioconversion of 7-β-xylosyl-10-deacetyltaxol or its extract was further optimized and scaled up with the engineered yeast harvested from 200-L scale high-cell-density fermentation. The optimization included the freeze-dried cell amount, dimethyl sulfoxide concentration, addition of 0.5 % antifoam supplement, and substrate concentration. A 93–95 % bioconversion and 83 % bioconversion of 10 and 15 g/L 7-β-xylosyltaxanes in 10 L reaction volume were achieved, respectively. The yield of 10-deacetyltaxol reached 10.58 g/L in 1 L volume with 15 g/L 7-β-xylosyl-10-deacetyltaxol. The conversion efficiencies were not only much higher than those of other reports and our previous work, but also realized in 10 L reaction volume. A pilot-scale product purification was also established. Our study bridges the gap between the basic research and commercial utilization of 7-β-xylosyl-10-deacetyltaxol for the industrial production of semi-synthetic paclitaxel.

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Acknowledgments

This work was supported by the National Natural Science Foundation (Grant Nos. 31270796; 30770229), the National Mega-project for Innovative Drugs (Grant No. 2012ZX09301002-001-005), and the Graduate Student Innovation Fund of Peking Union Medical College (Grant No. 2013-1007-02). The high-cell-density fermentation was performed in the Polo Biology Science Park Co., Ltd., China.

Conflict of interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines and Key Laboratory of Biosynthesis of Natural Products of National Health and Family Planning Commission, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, People’s Republic of China

    Wan-Cang Liu & Ping Zhu

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  1. Wan-Cang Liu
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  2. Ping Zhu
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Correspondence to Ping Zhu.

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10295_2015_1617_MOESM1_ESM.docx

Table S1 The impact of DMSO concentration on the bioconversion (reaction volume: 2 mL, n = 3, error value shows SD. For reaction conditions see the text) (DOCX 25 kb)

Fig. S1 The impact of some co-solvents or their binary solutions on the XDT bioconversion (DOCX 102 kb)

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Liu, WC., Zhu, P. Pilot studies on scale-up biocatalysis of 7-β-xylosyl-10-deacetyltaxol and its analogues by an engineered yeast. J Ind Microbiol Biotechnol 42, 867–876 (2015). https://doi.org/10.1007/s10295-015-1617-6

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  • DOI: https://doi.org/10.1007/s10295-015-1617-6

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