Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (3): 139-148.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0620

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Screening and Identification of Strains Producing 7-β-xylosyltaxanes Glycoside Hydrolases from the Periplaneta americana Gut

TANG Bin(), LIU Wen-bin, LI Xiao-bo, WANG Ning, JIN Xiao-bao()   

  1. College of Life Sciences and Biopharmaceutics,Guangdong Pharmaceutical University,Guangdong Key Laboratory of Pharmaceutical Bioactive Substances,Guangzhou 510006
  • Received:2021-05-11 Online:2022-03-26 Published:2022-04-06
  • Contact: JIN Xiao-bao E-mail:1375540061@qq.com;jinxf2001@163.com

Abstract:

The objective of this work is to screen endophytes that can produce 7-β-xylosyltaxanes glycoside hydrolases and to identify its biological characteristics. Taking total 178 strains of intestinal endophytes from Periplaneta americana preserved in the laboratory as the research object,the Congo red transparent circle method and fluorescent color method were used to screen the xylanase-producing strains,and the thin-layer chromatography was further used to screen strains producing the 7-β-xylosyltaxanes glycoside hydrolases. On the basis of this,a strain producing high 7-β-xylosyltaxanes glycoside hydrolases was selected for morphological,physiological,biochemical and molecular biological identification,and the conversion rate of 7-β-xylosyl-10-deacetylpaclitaxel by the strain was measured by high performance liquid chromatography. Among the 178 strains of intestinal endophytes in P. americana,total 55 endophytes produced xylanase,of which 8 strains of Streptomyces and 2 strains of Cupriavidus produced 7-β-xylosyltaxanes glycoside hydrolases. The high-yield strain WA11-2-9 was identified as Streptomyces enissocaesilis by bacterial morphology,physiological and biochemical experiments,combined with 16S rDNA sequence analysis. WA11-2-9 converted 7-β-xylosyl-10-deacetylpaclitaxel to 10-deacetylbaccatin III and 10-deacetylpaclitaxel. The conversion rate of the main product 10-deacetylpaclitaxel was 21.52%. In conclusion,the intestinal endophytes WA11-2-9(GenBank accession number:MZ411692)from P. americana secreted 7-β-xylosyltaxanes glycoside hydrolases,which lays a solid foundation for the subsequent biotransformation of paclitaxel.

Key words: Periplaneta americana gut, 7-β-xylosyltaxanes glycoside hydrolases, 7-β-xylosyl-10-deacetylpac-litaxel, 10-deacetylpaclitaxel