Difference between revisions of "Part:BBa K1403017:Design"

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===Design Notes===
 
===Design Notes===
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The purpose of this part is to biosynthesize acetoin using <i>E. coli</i>. The sequence was synthesized as a gBlock by IDT with the codon optimized <i>aldB</i> downstream of constitutive promoter <partinfo>BBa_J23108</partinfo> and a sRBS. Alpha-acetolactate decarboxylase transforms acetolactate to acetoin, which has a butter scent.
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Codon optimized for E.Coli adaptation using IDT's engine. The sRBS was generated by the Salis RBS calculator.
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We got the original L.Lactis aldB sequence from NCBI database. The sequence codes for acetolactate decarboxylase activity that transforms acetolactate produced by ilvBN gene, that original exists in E.Coli strain, to Acetoin, which produces butter smell. We then optimized the sequence. It is used for the production of butter smell for Paris-Bettencourt 2014 odor library project. It was transformed with the plasmid double expressing ilvBN gene, in order to raise basal metabolism of the pathway.
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The promoter was selected as a standard part for our odor library project, with a medium to high expression rate.
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Pathway: ilvBN produces acetolactate decarboxylase ----> aldB transforms acetolactate decarboxylase to acetoin
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===Source===
 
===Source===
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Non-redundant <i>Lactococcus lactis</i> AldB from [http://www.ncbi.nlm.nih.gov/protein/500160484?report=genbank&log$=protalign&blast_rank=1&RID=46AXW03K01R NCBI: WP_011835154.1].
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From L. lactis. Sequence from NCBI database.  
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===References===
 
===References===
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[https://parts.igem.org/Promoters/Catalog/Anderson Anderson Promoter Collection]
  
 
David R. Nielsen, Sang-Hwal Yoon, Clara J. Yuan and Kristala L. J. Prather, Metabolic engineering of acetoin and meso-2,3-butanediol biosynthesis in E. coli, January 2010.
 
David R. Nielsen, Sang-Hwal Yoon, Clara J. Yuan and Kristala L. J. Prather, Metabolic engineering of acetoin and meso-2,3-butanediol biosynthesis in E. coli, January 2010.
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[https://salis.psu.edu/software/forward_withConstraints  Salis lab RBS calculator]<br>
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H.M Salis, Methods in Enzymology 2011<br>
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H.M. Salis, E.A. Mirsky, C.A. Voigt, Nat. Biotech., 2009

Latest revision as of 07:39, 31 October 2014

Acetaldehyde dehydrogenase (aldB) expression cassette


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 71
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The purpose of this part is to biosynthesize acetoin using E. coli. The sequence was synthesized as a gBlock by IDT with the codon optimized aldB downstream of constitutive promoter BBa_J23108 and a sRBS. Alpha-acetolactate decarboxylase transforms acetolactate to acetoin, which has a butter scent.

Source

Non-redundant Lactococcus lactis AldB from [http://www.ncbi.nlm.nih.gov/protein/500160484?report=genbank&log$=protalign&blast_rank=1&RID=46AXW03K01R NCBI: WP_011835154.1].

References

Anderson Promoter Collection

David R. Nielsen, Sang-Hwal Yoon, Clara J. Yuan and Kristala L. J. Prather, Metabolic engineering of acetoin and meso-2,3-butanediol biosynthesis in E. coli, January 2010.

Salis lab RBS calculator
H.M Salis, Methods in Enzymology 2011
H.M. Salis, E.A. Mirsky, C.A. Voigt, Nat. Biotech., 2009