High-yield production of GDP-fucose for fucosylation of molecules

Information

  • Research Project
  • 8592879
  • ApplicationId
    8592879
  • Core Project Number
    R43GM106459
  • Full Project Number
    1R43GM106459-01A1
  • Serial Number
    106459
  • FOA Number
    PA-12-088
  • Sub Project Id
  • Project Start Date
    9/30/2013 - 11 years ago
  • Project End Date
    3/31/2014 - 10 years ago
  • Program Officer Name
    MARINO, PAMELA
  • Budget Start Date
    9/30/2013 - 11 years ago
  • Budget End Date
    3/31/2014 - 10 years ago
  • Fiscal Year
    2013
  • Support Year
    01
  • Suffix
    A1
  • Award Notice Date
    9/12/2013 - 11 years ago
Organizations

High-yield production of GDP-fucose for fucosylation of molecules

DESCRIPTION: Fucosylation plays an important role in much cellular process. Fucosylated oligosaccharides in the cell are involved in various types of biochemical recognition processes and in microbial infections, toxin entry, and cancer cells metastasis. These properties make these carbohydrates valuable for pharmaceutical and drug discovery needs but current production methods production is very expensive and impractical. Most notably is the expense and difficulty in producing the activated sugar, GDP-fucose. Current methods described to date for the production of GDP-fucose using chemoenzymatic synthesis, or modified microorganisms such E. coli and S. cerevisiae all either yield too small quantities of material or are overly complicated and can't be scaled. Here we propose to develop an entirely new system based on the use of a yeast system for production GDP-fucose. This system uses a strong inducible promoter for overexpression of the enzyme from a de novo pathway in methylotrophic yeast. These yeast naturally produce a high yield of GDP-mannose which the proposed system will convert to GDP-fucose and utilize nucleotide-sugar transporter for extracellular release. We foresee the advantages of this approach to be high- level expression of protein involved to GDP-fucose synthesis and transport and the possibility to use the desired enzymes for in vivo synthesis. In Phase I we will test the feasibility of developing this system by screening a set of yeast that can produce GDP-mannose in highest yield, overexpress the enzymes necessary to convert GDP-mannose to GDP-fucose, and test the ability to produce GDP-L-fucose. We will then test the ability to transport GDP-L- fucose out of the cell and determine the initial conditios for fermentation. In Phase II we will further engineer and optimize the production of GDP-fucose and demonstrate its utility by testing the production of several human milk from starting materials that are readily available to us. Finally, Phase III commercialization will involve sellng GDP-fucose, licensing the system for use by a variety of companies, and in using it to produce custom fucosylated oligosaccharides, small molecules, and proteins.

IC Name
NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
  • Activity
    R43
  • Administering IC
    GM
  • Application Type
    1
  • Direct Cost Amount
  • Indirect Cost Amount
  • Total Cost
    186972
  • Sub Project Total Cost
  • ARRA Funded
    False
  • CFDA Code
    859
  • Ed Inst. Type
  • Funding ICs
    NIGMS:186972\
  • Funding Mechanism
    SBIR-STTR RPGs
  • Study Section
    ZRG1
  • Study Section Name
    Special Emphasis Panel
  • Organization Name
    ZUCHEM, INC.
  • Organization Department
  • Organization DUNS
    126677041
  • Organization City
    CHICAGO
  • Organization State
    IL
  • Organization Country
    UNITED STATES
  • Organization Zip Code
    606124814
  • Organization District
    UNITED STATES