Part:BBa_K5327002
Methylthioalkylmalate synthase 1, chloroplastic
Determines the side chain length of aliphatic glucosinolate structures. Catalyzes exclusively the condensation reactions of both the first and second methionine carbon chain elongation.
Usage and Biology
Genome localization:Chromosome 5 - NC_003076.8
Expression diagram:
- Fig 1. The expression diagram of methylthioalkylmalate synthase 1, chloroplastic
Corresponding enzyme structure:
- Fig 2. The corresponding enzyme structure of methylthioalkylmalate synthase 1, chloroplastic
The PCR result:
- Fig 3. The PCR result of methylthioalkylmalate synthase 1, chloroplastic
Subcellular localization:[3]
Located in the chloroplast of cells
- Fig 4. The subcellular localization of methylthioalkylmalate synthase 1, chloroplastic
Dynamics data:
- Table 1. The dynamics data of methylthioalkylmalate synthase 1, chloroplastic
Design Notes
To obtain the key intermediate product dihomomethionine, we optimized the Methylthioalkylmalate synthase 1 (MAM1) gene from Arabidopsis thaliana and codon-optimized it for Saccharomyces cerevisiae (S288C) to ensure its efficient expression in yeast. MAM1 specifically catalyzes the first and second condensation reactions in the methionine carbon chain elongation, determining the side chain length of aliphatic glucosinolates. [4]We selected the CDC19 promoter (CDC19pBBa_K3772015) and ADH1 terminator (ADH1tBBa_K2637012) to drive and stabilize gene expression. After designing the optimized gene, it was inserted into a vector and integrated into yeast through homologous recombination. Defective strains were then used for screening and expression verification. This strategy ensures that MAM1 efficiently catalyzes methionine chain elongation, particularly in producing the key intermediate dihomomethionine, thereby facilitating the synthesis of the desired product.
Plasmid
- Fig 5. The plasmid expression of methylthioalkylmalate synthase 1, chloroplastic
Source
Arabidopsis thaliana
References
- ↑ TEXTOR S, BARTRAM S, KROYMANN J, et al. Biosynthesis of methionine-derived glucosinolates in Arabidopsis thaliana: recombinant expression and characterization of methylthioalkylmalate synthase, the condensing enzyme of the chain-elongation cycle [J]. Planta, 2004, 218(6): 1026-35.
- ↑ BENDEROTH M, TEXTOR S, WINDSOR A J, et al. Positive selection driving diversification in plant secondary metabolism [J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(24): 9118-23.
- ↑ https://www.uniprot.org/uniprotkb/Q9FG67/entry
- ↑ TEXTOR S, BARTRAM S, KROYMANN J R, et al. Biosynthesis of methionine-derived glucosinolates in Arabidopsis thaliana : recombinant expression and characterization of methylthioalkylmalate synthase, the condensing enzyme of the chain-elongation cycle [J]. Planta, 2004, 218(6): 1026-35.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 907
- 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 907
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 907
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 907
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 283
Illegal BsaI.rc site found at 472
Illegal SapI.rc site found at 591
None |