title: Bioproduction of baccatin III, an advanced precursor of paclitaxol, with transgenic Flammulina velutipes expressing the 10-deacetylbaccatin III-10-O-acetyl transferase gene.
abstract: BACKGROUND
  10-Deacetylbaccatin III (10-DAB) and baccatin III are intermediates in the biosynthesis of Taxol (an anti-cancer drug) and useful precursors for semi-synthesis of the drug. In this study, a bioconversion system was established for the production of baccatin III, an advanced precursor of paclitaxel, in the transgenic mushroom Flammulina velutipes expressing the 10-deacetylbaccatin III-10β-O-acetyltransferase gene. The expression vector pgFvs-TcDBAT containing the 10-deacetylbaccatin III-10β-O-acetyltransferase (DBAT) gene was constructed and transformed into the cells of F. velutipes by polyethylene glycol-mediated protoplast transformation.
  
  
  RESULTS
  Polymerase chain reaction and Southern blotting analysis verified the successful integration of the exogenous DBAT gene into the genome of F. velutipes. Reverse transcription polymerase chain reaction and enzyme activity analyses confirmed that the DBAT gene was expressed in F. velutipes, and DBAT is able to convert substrate into baccatin III.
  
  
  CONCLUSIONS
  The DBAT gene from the plant Taxus chinensis can be functionally expressed in F. velutipes. Transgenic F. velutipes expressing the DBAT gene is able to produce the target product, baccatin III. This is the first report about the transformation and expression of paclitaxel biosynthetic gene in the edible mushroom F. velutipes. This represents a significant step towards bio-production of paclitaxel and its advanced precursor baccatin III in an edible fungus.
authors.affiliation: Department of Bioengineering, College of Food Science, South China Agriculture University, Guangzhou, Guangdong, 510640, China; Guangdong Shantou Institute of Quality and Metrology Supervision Testing, Shantou, 515041, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Han
authors.forename: Lin-Zhi
authors.initials: LZ
authors.lastname: Kang
authors.forename: Xian-Lu
authors.initials: XL
authors.lastname: Zeng
authors.forename: Zhi-Wei
authors.initials: ZW
authors.lastname: Ye
authors.forename: Li-Qiong
authors.initials: LQ
authors.lastname: Guo
authors.forename: Jun-Fang
authors.initials: JF
authors.lastname: Lin
mesh_terms: D000123:Acetyltransferases / Q000235:genetics* / Q000378:metabolism; D000470:Alkaloids / Q000096:biosynthesis*; D055393:Flammulina / Q000235:genetics* / Q000378:metabolism; D017343:Genes, Plant; D030781:Organisms, Genetically Modified; D017239:Paclitaxel / Q000096:biosynthesis*; D043823:Taxoids / Q000378:metabolism*; D020946:Taxus / Q000201:enzymology / Q000235:genetics*
chemical_list: D000470:Alkaloids; D043823:Taxoids; C073950:baccatin III; C042213:10-deacetylbaccatine III; D000123:Acetyltransferases; D017239:Paclitaxel
keywords: 10-deacetylbaccatin III-10β-O-acetyltransferase; Flammulina velutipes; baccatin III; bioconversion; paclitaxel