Team:KULeuven/Notebook/Vanillin Production

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Revision as of 13:42, 27 July 2009 by Deepstar (Talk | contribs)


13 July

  • Vanillin synthesis: DNA not yet complete/in library [http://partsregistry.org/wiki/index.php/Part:BBa_I742140]
    • Combination of 5 genes; 3 ervan zijn beschikbaar
      • [http://parts.mit.edu/igem07/index.php/Edinburgh/Yoghurt/Design#Vanilla_Flavour_Production Edinburgh Vanilla production] (Synthese stappen door Edinburgh igem 2007)
    • [http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17437627#id538997 Vanillin production using metabolically engineered Escherichia coli under non-growing conditions], Ferrulic accid --> Vanillin, 5k bp

14 July

  • Biobricks for vanillin synthesis
    • Send mail to University of Tuscia [Done]
    • Order:
      • (sam8 (tyrosine-ammonia lyase) coding sequence) [ordered]
      • (sam5 (coumarate hydroxylase) coding sequence) [ordered]
      • (COMT gene with ribosome binding site) [Available in kit]

Extra info on Vanillin synthesis

http://www.ncbi.nlm.nih.gov/pubmed/16232583 1 B C Okeke and V Venturi ,Construction of recombinants Pseudomonas putida BO14 and Escherichia coli QEFCA8 for ferulic acid biotransformation to vanillin, J Biosci Bioeng. 1999; 88(1): 103–106.
http://www.ncbi.nlm.nih.gov/pubmed/14602615 2 Jörg Overhage, Alexander Steinbüchel, and Horst Priefert,Highly Efficient Biotransformation of Eugenol to Ferulic Acid and Further Conversion to Vanillin in Recombinant Strains of Escherichia coli, Appl Environ Microbiol. 2003 November; 69(11): 6569–6576. doi: 10.1128/AEM.69.11.6569-6576.2003.
http://www.ncbi.nlm.nih.gov/pubmed/10616715 3 J Overhage, H Priefert, J Rabenhorst, and A Steinbüchel,Biotransformation of eugenol to vanillin by a mutant of Pseudomonas sp. strain HR199 constructed by disruption of the vanillin dehydrogenase (vdh) gene, Appl Microbiol Biotechnol. 1999 November; 52(6): 820–828.
http://www.labmeeting.com/paper/2776259/yoon-enhanced-vanillin-production-from-recombinant-e-coli-using-ntg-mutagenesis-and-adsorbent-resin 4 Yoon SH, Lee EG, Das A, Lee SH, Li C, Ryu HK, Choi MS, Seo WT, and Kim SW, Enhanced vanillin production from recombinant E. coli using NTG mutagenesis and adsorbent resin, Biotechnology progress 23(5):1143.
http://www.scielo.br/scielo.php?pid=S1517-83822003000500037&script=sci_arttext 5 Attilio ConvertiI; Danilo de FaveriI; Patrizia PeregoI; Paolo BarghiniII; Maurizio RuzziII; Luciane SeneIII, Vanillin production by recombinant strains of Escherichia coli, Braz. J. Microbiol. vol.34 suppl.1 São Paulo Nov. 2003
http://www.springerlink.com/content/bj47xnqn5kr65wvp/fulltext.pdf 6 S. Achterholt, H. Priefert, and A. Steinbüchel, Identification of Amycolatopsis sp. strain HR167 genes, involved in the bioconversion of ferulic acid to vanillin, Appl Microbiol Biotechnol. 2000 December; 54(6): 799–807

16 July

  • Focus on the last two parts in the synthesis of vanillin since that pathway has been proven
    • Can always add additional 3 paths
  • Send mail to U. Edinburgh for the proteins for vanillin synthesis [Done]
    • Received mail back from Edinburgh, biobricks of the parts should be done. Will receive answer next week

24 July

Received mail from U. Edinburgh: We will receive:

  • Sam5; no part number, Pst1 site removed
  • Sam8; coding sequence without RBS
  • ComT; coding sequence without RBS
  • ech; RBS + ech, in pSB1A2,
  • fcs; RBS + fcs, in pSB1A2,


Week 1: 6 July 2009 - 12 July 2009

Week 2: 13 July 2009 - 19 July 2009

[edit] Monday

  • Vanillin synthesis: DNA not yet complete/in library [http://partsregistry.org/wiki/index.php/Part:BBa_I742140]
    • Combination of 5 genes; 3 are available
    • [http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17437627#id538997 Vanillin production using metabolically engineered Escherichia coli under non-growing conditions], Ferulic accid --> Vanillin, 5k bp

[edit] Tuesday

  • Biobricks for vanillin synthesis
    • Send mail to University of Tuscia [Done]
    • Order:
      • (sam8 (tyrosine-ammonia lyase) coding sequence) [ordered]
      • (sam5 (coumarate hydroxylase) coding sequence) [ordered]
      • (COMT gene with ribosome binding site) [Available in kit]

Extra info on vanillin synthesis

[edit] Thursday

  • Focus on the last two parts in the synthesis of vanillin since that pathway has been proven
    • Can always add additional 3 paths
  • Send mail to U. Edinburgh for the proteins for vanillin synthesis [Done]
    • Received mail back from Edinburgh, biobricking of the parts should still be done. Will receive answer next week

Week 3: 20 July 2009 - 26 July 2009

[edit] Friday

Received mail from U. Edinburgh: We will receive:

  • sam5; no part number, Pst1 site removed
  • sam8; coding sequence without RBS
  • COMT; coding sequence without RBS
  • ech; RBS + ech, in ,
  • fcs; RBS + fcs, in ,

Week 4: 27 July 2009 - 2 August 2009

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