
Part:BBa_K849000
3-Hydroxy-3-Methyl-Glutaryl-CoA-Reductase from Yeast
This is a truncated version of the 3-Hydroxy-3-Methyl-Glutaryl-Coenzyme-A-Reductase (HMG-CoA-R) from Saccharomyces cerevisiae. It catalyses the reaction of 3-Hydroxy-3-Methyl-Glutaryle-Coenzyme-A to Mevalonate which occurs in the Mevalonate Pathway to Isopentylpyrophosphate which is the Precurser of all Isoprenoides. The enzyme is naturally a membrane bound enzyme. The membrane anchor is situated N-terminal. This region is also essential for the degradational control of the enzyme. By deleting the N-terminal part to Aminoacid (inclusive) the enzyme is expected to be water soluble and deregulated.
Trunctuation had been done according to the following publication: http://aem.asm.org/content/63/9/3341.short
K A Donald, R Y Hampton and I B Fritz: Effects of overproduction of the catalytic domain of 3-hydroxy-3-methylglutaryl coenzyme A reductase on squalene synthesis in Saccharomyces cerevisiae . Appl. Environ. Microbiol. 1997, 63(9):3341.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 685
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1456
- 1000COMPATIBLE WITH RFC[1000]
Characterization-Team Nagahama 2017-

HMG1 encodes HMG-CoA reductase and catalyzes conversion of HMG-CoA to mevalonate, which is a rate-limiting step in sterol biosynthesis.
Because mevalonate is synthesized, it is easy to flow to subsequent pathways.
Therefore,β-carotene production increase.
Color of colony was changed yellow from orange when tHMG1 was inserted into S. cerevisiae synthesizing carotenoid.(Fig.2,3,4)
This result showed that tHMG1 was inserted.
tHMG1 has a characteristic of changing color of S. cerevisiae. This result to be due to the fact that as a result of massive synthesis of mevalonate, β-carotene was produced in large amount.
In order to investigate whether tHMG1 increased β-carotene productions, color of S. cerevisiae and β-carotene were compared.
It is considered that the amount of β-carotene synthesized increased because the color of yeast introduced tHMG1 was closed the color of 1μg of β-carotene dissolved in 1 ml of hexane.
S. cerevisiae transformed was grown onto sd-ura medium. This figure shows that the pYES2 plasmid inserted sod 2 was introduced into S. cerevisiae.
Reference
[1]Verwaal R, Wang J, Meijnen JP, Visser H, Sandmann G, van den Berg JA, van Ooyen AJ (2007) High-level production of beta-carotene in Saccharomyces cerevisiae by successive transformation with carotenogenic genes from Xanthophyllomyces dendrorhous. Appl Environ Microbiol 73(13):4342–4350
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