search
for
 About Bioline  All Journals  Testimonials  Membership  News


Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 30, No. 1, 2017, pp. 1-5
Bioline Code: ej17069
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 30, No. 1, 2017, pp. 1-5

 en Introduction of a synthetic Thermococcus check for this species in other resources -derived α-amlyase gene into barley genome for increased enzyme thermostability in grains
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.

Keywords
Amylopectin; Amylose; Beer; Fermentation; Gene transfer; Glutenin; Hordeum; Promoter; Seed-directed expression; Synthetic gene; Thermotolerant bacteria

 
© Copyright 2017 - Pontificia Universidad Católica de Valparaíso
Alternative site location: http://www.ejbiotechnology.info

Home Faq Resources Email Bioline
© Bioline International, 1989 - 2024, Site last up-dated on 01-Sep-2022.
Site created and maintained by the Reference Center on Environmental Information, CRIA, Brazil
System hosted by the Google Cloud Platform, GCP, Brazil