Part:BBa_K4180002

snf1Δ2-306aa N-truncated

snf1Δ2-306aa, N-terminus truncated from amino acid 2 to 306 deleting kinase domain of SNF1, where the phosphorylation takes place on Thr210 to activate the catalytic activity of SNF1

Citation: McCartney, R R, and M C Schmidt. “Regulation of Snf1 kinase. Activation requires phosphorylation of threonine 210 by an upstream kinase as well as a distinct step mediated by the Snf4 subunit.” The Journal of biological chemistry vol. 276,39 (2001): 36460-6. doi:10.1074/jbc.M104418200

Sequence and Features

        BBa K4180002
        snf1Δ2-306aa
         PCR product

After Bacteria transformation, bacteria colonies were picking up directly to do snf1Δ2-306aa - colonies had BBa_K4180002 snf1Δ2-306aa
         PCR product

Contribution From NJXDF-CHN 2022

Group: NJXDF-CHN

Author: Yang Gu

Summary: Charactering the effect of the SNF1 gene for lipid synthesis and growth in Y.lipolytica.

Characterization from iGEM22-NJXDF-CHN

         Sucrose non-fermenting (Snf1) protein kinase, the yeast ortholog of mammalian AMP-activated protein kinase, is the key component of the glucose repression pathway and the energy sensor in yeast. When the ratio of adenosine monophosphate (AMP) to adenosine triphosphate (ATP) is high, it increased catabolic processes to obtain more ATP and decreases anabolic processes to reduce energy consumption. When truncated from the N-terminal amino acid 2 to amino acid 306, the kinase structure of SNF1 can be deleted and the protein inactivated(Liu et al. 2020). It was first registered in 2022. In order to characterize the effect of the IDH2 (BBa_K4297072) gene in lipid synthesis and growth, we constructed a knockout strain po1f ΔylSNF1 using homologous recombination and tested for fermentation in YNB mediums. The experimental results show that the strain po1f ΔylSNF1 can effectively increase the accumulation of biomass and fatty acid content in YNB medium, up to 83.77% and 159.68% at 120h, respectively (Fig.3). These results provide references for future iGEM teams to improve biomass and lipid accumulation in Y.lipolytica.

Fig. 3. The growth and lipid synthesis of strain po1f ΔylSNF1 in YNB medium. A. Changes of growth curves in YNB medium. B. Changes of fatty acid content in YNB medium at 120h.

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]