Part:BBa_K4845009

temA

temA

Basic Part BBa_K4845009 (temA)

Profile

• Name: temA
• Base Pairs: 1869 bp
• Origin: Synthetic
• Properties: Glucoamylase (α-1,4-Glucan glucohydrolase) is a key exoglycosidase for alcohol fermentation of starch raw materials. It can sequentially hydrolyze α-1,4 glycosidic bonds from the non-reducing ends of starch and cut off each glucose unit. When encountering the branching point of amylopectin, it can also hydrolyze the α-1,6 glycosidic bond, thereby completely hydrolyzing amylopectin into glucose.

Usage and Biology

Innis MA et al. proposed that the yeast's own enolase gene promoter and the glucoamylase gene of Aspergillus awamori were integrated into the pAC1 plasmid, and the plasmid was introduced into the Saccharomyces cerevisiae cell to realize the expression of the saccharifying enzyme gene in the yeast, so that the recombinant Saccharomyces cerevisiae can grow normally with starch as the sole carbon source¹.

Ashikari et al. assumed that the modified Rhizopus glucoamylase gene was introduced into Saccharomyces cerevisiae, and the ungelatinized corn starch was directly fermented by saccharifying yeast to obtain about 13% (v/v) ethanol. In non-cooking fermentation, these transformants will help to reduce the amount of enzyme required, thereby reducing the cost of alcohol production².

Li Wenqing et al. obtained the glucoamylase gene cDNA of Aspergillus niger without 5' non-coding region by PCR technology and inserted the cDNA sequence between the promoter and terminator of the PGK gene, which reduced the effect of RNA secondary structure formed by the 5' non-coding region on the expression of the glucoamylase gene. It was recombined into pMAG17 integrated plasmid and transformed into S. cerevisiae, which improved the expression and secretion of the glucoamylase gene³.

Reference

1. Innis MA, Holland MJ, McCabe PC, Cole GE, Wittman VP, Tal R, Watt KW, Gelfand DH, Holland JP, Meade JH (1985) Expression, glycosylation, and secretion of an Aspergillus Glucoamylase by Saccharomyces cerevisiae. Science 228(4695):21–26.
2. Ashkari T, Nakamura N, Tanaka Y, et al. Rhizopus Raw-Starch-Degrading Glucoamylase: Its Cloning and Expression in Yeast. Agricultural and Biological Chemistry, 2014, 50(4).
3. Wu Xiaoping, Li Wenqing, Luo Jinxian. Expression of α-amylase and glucoamylase and construction of engineered Saccharomyces cerevisiae. Journal of Sun Yat-sen University (Natural Science Edition), 1999(02): 81-85.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 1382
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1619
  
• 1000
  COMPATIBLE WITH RFC[1000]