Introduction of a synthetic Thermococcus-derived α-amlyase gene into barley genome for increased enzyme thermostability in grains
Electron. j. biotechnol
;
30: 1-5, nov. 2017. ilus, tab, graf
Artículo
en Inglés
| LILACS
| ID: biblio-1021034
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 7585°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.
Texto completo:
Disponible
Índice:
LILACS (Américas)
Asunto principal:
Semillas
/
Hordeum
/
Thermococcus
/
Alfa-Amilasas
Idioma:
Inglés
Revista:
Electron. j. biotechnol
Asunto de la revista:
Biotecnologia
Año:
2017
Tipo del documento:
Artículo
/
Documento de proyecto
País de afiliación:
Eslovaquia
Institución/País de afiliación:
National Agricultural and Food Centre/SK
/
Plant Science and Biodiversity Center SAS/SK
/
Slovak Academy of Sciences/SK
/
University of Ss. Cyril and Methodius/SK
/
University of Ss., Cyril and Methodius in Trnava/SK
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