Cloning, heterologous expression, and enzymatic characterization of a thermostable glucoamylase from Talaromyces emersonii

Protein Expr Purif. 2002 Oct;26(1):1-8. doi: 10.1016/s1046-5928(02)00505-3.

Abstract

The gene encoding a thermostable glucoamylase from Talaromyces emersonii was cloned and, subsequently, heterologously expressed in Aspergillus niger. This glucoamylase gene encodes a 618 amino acid long protein with a calculated molecular weight of 62,827Da. T. emersonii glucoamylase fall into glucoside hydrolase family 15, showing approximately 60% sequence similarity to glucoamylase from A. niger. The expressed enzyme shows high specific activity towards maltose, isomaltose, and maltoheptaose, having 3-6-fold elevated k(cat) compared to A. niger glucoamylase. T. emersonii glucoamylase showed significantly improved thermostability with a half life of 48h at 65 degrees C in 30% (w/v) glucose, compared to 10h for glucoamylase from A. niger. The ability of the glucoamylase to hydrolyse amylopectin at 65 degrees C is improved compared to A. niger glucoamylase, giving a significant higher final glucose yield at elevated temperatures. The increased thermal stability is thus reflected in the industrial performance, allowing T. emersonii glucoamylase to operate at a temperature higher than the A. niger enzyme.

MeSH terms

  • Aspergillus niger / genetics*
  • Cloning, Molecular
  • Enzyme Stability
  • Gene Expression
  • Glucan 1,4-alpha-Glucosidase / chemistry
  • Glucan 1,4-alpha-Glucosidase / genetics*
  • Glucan 1,4-alpha-Glucosidase / metabolism*
  • Glucans / metabolism
  • Hydrogen-Ion Concentration
  • Isomaltose / metabolism
  • Kinetics
  • Maltose / metabolism
  • Molecular Sequence Data
  • Recombinant Fusion Proteins
  • Substrate Specificity
  • Talaromyces / enzymology*
  • Talaromyces / genetics
  • Temperature

Substances

  • Glucans
  • Recombinant Fusion Proteins
  • maltoheptaose
  • Isomaltose
  • Maltose
  • Glucan 1,4-alpha-Glucosidase

Associated data

  • GENBANK/AJ304803