A hyper-thermostable α-amylase from Pyrococcus furiosus accumulates in Nicotiana tabacum as functional aggregates.
| Autor: | Zhu H; Agriculture and Agri-Food Canada, London Research and Development Centre, London, Ontario, Canada., Reynolds LB; Agriculture and Agri-Food Canada, London Research and Development Centre, London, Ontario, Canada., Menassa R; Agriculture and Agri-Food Canada, London Research and Development Centre, London, Ontario, Canada. rima.menassa@agr.gc.ca.; Department of Biology, University of Western Ontario, London, Ontario, Canada. rima.menassa@agr.gc.ca. |
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| Jazyk: | angličtina |
| Zdroj: | BMC biotechnology [BMC Biotechnol] 2017 Jun 19; Vol. 17 (1), pp. 53. Date of Electronic Publication: 2017 Jun 19. |
| DOI: | 10.1186/s12896-017-0372-3 |
| Abstrakt: | Background: Alpha amylase hydrolyzes α-bonds of polysaccharides such as starch and produces malto-oligosaccharides. Its starch saccharification applications make it an essential enzyme in the textile, food and brewing industries. Commercially available α-amylase is mostly produced from Bacillus or Aspergillus. A hyper-thermostable and Ca 2++ independent α-amylase from Pyrococcus furiosus (PFA) expressed in E.coli forms insoluble inclusion bodies and thus is not feasible for industrial applications. Results: We expressed PFA in Nicotiana tabacum and found that plant-produced PFA forms functional aggregates with an accumulation level up to 3.4 g/kg FW (fresh weight) in field conditions. The aggregates are functional without requiring refolding and therefore have potential to be applied as homogenized plant tissue without extraction or purification. PFA can also be extracted from plant tissue upon dissolution in a mild reducing buffer containing SDS. Like the enzyme produced in P. furiosus and in E. coli, plant produced PFA preserves hyper-thermophilicity and hyper-thermostability and has a long shelf life when stored in lyophilized leaf tissue. With tobacco's large biomass and high yield, hyper-thermostable α-amylase was produced at a scale of 42 kg per hectare. Conclusions: Tobacco may be a suitable bioreactor for industrial production of active hyperthermostable alpha amylase. |
| Databáze: | MEDLINE |
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