Cloning, expression, and purification of insect ( Sitophilus oryzae ) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host
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| 024 | 7 | 0 | |a 10.1007/s00253-014-6314-2 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-014-6314-2 | ||
| 245 | 0 | 0 | |a Cloning, expression, and purification of insect ( Sitophilus oryzae ) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host |h [Elektronische Daten] |c [Ewelina Celińska, Wojciech Białas, Monika Borkowska, Włodzimierz Grajek] |
| 520 | 3 | |a Raw-starch-digesting enzymes (RSDE) are of major importance for industrial applications, as their usage greatly simplifies the starch processing pipeline. To date, only microbial RSDE have gained considerable attention, since only microbial production of enzymes meets industrial demands. In this study, α-amylase from rice weevil (Sitophilus oryzae), the major rice pest, was cloned and expressed in Yarrowia lipolytica Po1g strain. The enzyme was secreted into the culture medium, and the peak activity (81AU/L) was reached after only 29h of culturing in 5-L bioreactors. Through simple purification procedure of ammonium sulfate precipitation and affinity chromatography, it was possible to purify the enzyme to apparent homogeneity (25-fold purification factor, at 5% yield). The optimal conditions for the α-amylase activity were pH5.0 and a temperature of 40°C. The α-amylase studied here did not show any obligate requirement for Ca2+ ions. The recombinant α-amylase appeared to efficiently digest granular starch from pea, amaranth, waxy corn, and waxy rice. | |
| 540 | |a The Author(s), 2014 | ||
| 690 | 7 | |a Amylase |2 nationallicence | |
| 690 | 7 | |a Granular starch |2 nationallicence | |
| 690 | 7 | |a Rice weevil |2 nationallicence | |
| 690 | 7 | |a Yarrowia lipolytica |2 nationallicence | |
| 690 | 7 | |a Recombinant protein expression |2 nationallicence | |
| 700 | 1 | |a Celińska |D Ewelina |u Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, ul. Wojska Polskiego 48, 60-627, Poznań, Poland |4 aut | |
| 700 | 1 | |a Białas |D Wojciech |u Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, ul. Wojska Polskiego 48, 60-627, Poznań, Poland |4 aut | |
| 700 | 1 | |a Borkowska |D Monika |u Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, ul. Wojska Polskiego 48, 60-627, Poznań, Poland |4 aut | |
| 700 | 1 | |a Grajek |D Włodzimierz |u Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, ul. Wojska Polskiego 48, 60-627, Poznań, Poland |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/6(2015-03-01), 2727-2739 |x 0175-7598 |q 99:6<2727 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-014-6314-2 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-014-6314-2 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Celińska |D Ewelina |u Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, ul. Wojska Polskiego 48, 60-627, Poznań, Poland |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Białas |D Wojciech |u Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, ul. Wojska Polskiego 48, 60-627, Poznań, Poland |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Borkowska |D Monika |u Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, ul. Wojska Polskiego 48, 60-627, Poznań, Poland |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Grajek |D Włodzimierz |u Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, ul. Wojska Polskiego 48, 60-627, Poznań, Poland |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/6(2015-03-01), 2727-2739 |x 0175-7598 |q 99:6<2727 |1 2015 |2 99 |o 253 | ||