1. Amylomaltase from Thermus filiformis: expression in Saccharomyces cerevisiae and its use in starch modification
- Author
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Santhana Nakapong, Pitchanan Nimpiboon, Piamsook Pongsawasdi, and Suthipapun Tumhom
- Subjects
Manihot ,Starch ,Saccharomyces cerevisiae ,Applied Microbiology and Biotechnology ,Modified starch ,03 medical and health sciences ,chemistry.chemical_compound ,Bacterial Proteins ,Amylose ,Generally recognized as safe ,Food science ,Thermus ,030304 developmental biology ,0303 health sciences ,Cyclodextrins ,biology ,030306 microbiology ,Chemistry ,Peas ,Temperature ,food and beverages ,Glycogen Debranching Enzyme System ,General Medicine ,TFAM ,biology.organism_classification ,Yeast ,Recombinant Proteins ,Galactose ,Biotechnology - Abstract
Aim To express amylomaltase from Thermus filiformis (TfAM) in a generally recognized as safe (GRAS) organism and to use the enzyme in starch modification. Methods and results TfAM was expressed in Saccharomyces cerevisiae, using 2% (w/v) galactose inducer under GAL1 promoter. The enzyme was thermostable with high disproportionation and cyclization activities. The main large-ring cyclodextrin (CD) products were CD24-CD29, with CD26 as maximum at all incubation times. TfAM was used to modify cassava and pea starches, the amylose content decreased 18% and 30%, respectively, when 5% (w/v) starch was treated with 0·5 U TfAM g-1 starch. The increase in short branched chain (DP, degree of polymerization, 1-5) and the broader chain length distribution pattern which extended to the longer chain (DP40) after TfAM treatment were observed. The thermal property was changed, with an increase in retrogradation of starch as suggested by a lower enthalpy. Conclusions TfAM was successfully expressed in S. cerevisiae and was used to make starches with new functionality. Significance and impact of the study This is the first report on the expression of AM in the GRAS yeast and the production of a modified starch gel from pea starch to improve the versatility of starch for food use.
- Published
- 2019