We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Introduction of a synthetic Thermococcus-derived α-amlyase gene into barley genome for increased enzyme thermostability in grains.
- Authors
Mihálik, Daniel; Gubišová, Marcela; Kraic, Ján; Hudcovicová, Martina; Havrlentová, Michaela; Moravčíková, Jana; Glasa, Miroslav; Matušíková, Ildikó
- Abstract
Background: The enzymes utilized in the process of beer production are generally sensitive to higher temperatures. About 60% of them are deactivated in drying the malt that limits the utilization of starting material in the fermentation process. Gene transfer from thermophilic bacteria is a promising tool for producing barley grains harboring thermotolerant enzymes. Results: Gene for α-amylase from hydrothermal Thermococcus, optimally active at 75-85°C and pH between 5.0 and 5.5, was adapted in silico to barley codon usage. The corresponding sequence was put under control of the endosperm-specific promoter 1Dx5 and after synthesis and cloning transferred into barley by biolistics. In addition to model cultivar Golden Promise we transformed three Slovak barley cultivars Pribina, Levan and Nitran, and transgenic plants were obtained. Expression of the ~50 kDa active recombinant enzyme in grains of cvs. Pribina and Nitran resulted in retaining up to 9.39% of enzyme activity upon heating to 75°C, which is more than 4 times higher compared to non-transgenic controls. In the model cv. Golden Promise the grain α-amylase activity upon heating was above 9% either, however, the effects of the introduced enzyme were less pronounced (only 1.22 fold difference compared with non-transgenic barley). Conclusions: Expression of the synthetic gene in barley enhanced the residual α-amylase activity in grains at high temperatures.
- Subjects
BARLEY genetics; GENETIC transformation; BIOLISTICS; PLANT cloning; BARLEY varieties
- Publication
Electronic Journal of Biotechnology, 2017, Vol 30, p1
- ISSN
0717-3458
- Publication type
Article
- DOI
10.1016/j.ejbt.2017.08.002