Thermoseparating aqueous two phase system for the separation of alkaline proteases from fish viscera
The objective of this study was to isolate alkaline proteases from farmed giant catfish viscera by using a thermoseparating aqueous two phase system (T-ATPS). Different phase partitioning parameters [type of salts, NaCl addition and temperature were optimized. The optimum system contained 40% EOPO3900-10% MgSO4 with 17%NaCl, induced phase separation at 55ºC and provided the highest recovery (77.98 %) and purity (PF: 21.50-fold) with a partition coefficient (KE) of 11.90 and a volume ratio (VR) of 0.19. In recycling step, the mixture of bottom salt-rich phase and bottom EOPO-rich phase in the ratio of 0.5:1.5 (w/w) provided the highest recovery and purity. Hence, the total recovery of 91.62% was obtained from the separation system. Three major clear zones (24, 36 and 130 kDa) can be distinctively observed on casein-substrate gel electrophoresis for protease activity staining. Major protein components of perilla seed hydrolysates were completely hydrolyzed by the alkaline proteases at 70 units while those of red kidney bean isolate were more resistant to protein hydrolysis. Fifty units of Flavourzyme® hydrolyzed completely both plant proteins into small peptides or amino acids. As a result, TATPS could be used as an alternative method for separation of enzymes from various sources with acceptable recovery. Besides, the obtained alkaline proteases can be further used in preparation of protein hydrolysates.
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Format: | artículo biblioteca |
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Marcel Dekker
2014
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Subjects: | Recovery, Protein hydrolysates, Thermoseparating polymer, Viscera, Aqueous two-phase systems, Alkaline proteases, |
Online Access: | http://hdl.handle.net/10261/112395 |
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dig-ictan-es-10261-1123952019-02-01T08:23:23Z Thermoseparating aqueous two phase system for the separation of alkaline proteases from fish viscera Ketnawa, Sunantha Benjakul, Soottawat Martínez Álvarez, Óscar Rawdkuen, Saroat Recovery Protein hydrolysates Thermoseparating polymer Viscera Aqueous two-phase systems Alkaline proteases The objective of this study was to isolate alkaline proteases from farmed giant catfish viscera by using a thermoseparating aqueous two phase system (T-ATPS). Different phase partitioning parameters [type of salts, NaCl addition and temperature were optimized. The optimum system contained 40% EOPO3900-10% MgSO4 with 17%NaCl, induced phase separation at 55ºC and provided the highest recovery (77.98 %) and purity (PF: 21.50-fold) with a partition coefficient (KE) of 11.90 and a volume ratio (VR) of 0.19. In recycling step, the mixture of bottom salt-rich phase and bottom EOPO-rich phase in the ratio of 0.5:1.5 (w/w) provided the highest recovery and purity. Hence, the total recovery of 91.62% was obtained from the separation system. Three major clear zones (24, 36 and 130 kDa) can be distinctively observed on casein-substrate gel electrophoresis for protease activity staining. Major protein components of perilla seed hydrolysates were completely hydrolyzed by the alkaline proteases at 70 units while those of red kidney bean isolate were more resistant to protein hydrolysis. Fifty units of Flavourzyme® hydrolyzed completely both plant proteins into small peptides or amino acids. As a result, TATPS could be used as an alternative method for separation of enzymes from various sources with acceptable recovery. Besides, the obtained alkaline proteases can be further used in preparation of protein hydrolysates. Peer Reviewed 2015-03-16T10:27:43Z 2015-03-16T10:27:43Z 2014 2015-03-16T10:27:43Z artículo http://purl.org/coar/resource_type/c_6501 doi: 10.1080/01496395.2014.919595 issn: 0149-6395 Separation Science and Technology 49: 2158- 2168 (2014) http://hdl.handle.net/10261/112395 10.1080/01496395.2014.919595 none Marcel Dekker |
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Recovery Protein hydrolysates Thermoseparating polymer Viscera Aqueous two-phase systems Alkaline proteases Recovery Protein hydrolysates Thermoseparating polymer Viscera Aqueous two-phase systems Alkaline proteases |
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Recovery Protein hydrolysates Thermoseparating polymer Viscera Aqueous two-phase systems Alkaline proteases Recovery Protein hydrolysates Thermoseparating polymer Viscera Aqueous two-phase systems Alkaline proteases Ketnawa, Sunantha Benjakul, Soottawat Martínez Álvarez, Óscar Rawdkuen, Saroat Thermoseparating aqueous two phase system for the separation of alkaline proteases from fish viscera |
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The objective of this study was to isolate alkaline proteases from farmed giant catfish viscera by using a thermoseparating aqueous two phase system (T-ATPS). Different phase partitioning parameters [type of salts, NaCl addition and temperature were optimized. The optimum system contained 40% EOPO3900-10% MgSO4 with 17%NaCl, induced phase separation at 55ºC and provided the highest recovery (77.98 %) and purity (PF: 21.50-fold) with a partition coefficient (KE) of 11.90 and a volume ratio (VR) of 0.19. In recycling step, the mixture of bottom salt-rich phase and bottom EOPO-rich phase in the ratio of 0.5:1.5 (w/w) provided the highest recovery and purity. Hence, the total recovery of 91.62% was obtained from the separation system. Three major clear zones (24, 36 and 130 kDa) can be distinctively observed on casein-substrate gel electrophoresis for protease activity staining. Major protein components of perilla seed hydrolysates were completely hydrolyzed by the alkaline proteases at 70 units while those of red kidney bean isolate were more resistant to protein hydrolysis. Fifty units of Flavourzyme® hydrolyzed completely both plant proteins into small peptides or amino acids. As a result, TATPS could be used as an alternative method for separation of enzymes from various sources with acceptable recovery. Besides, the obtained alkaline proteases can be further used in preparation of protein hydrolysates. |
format |
artículo |
topic_facet |
Recovery Protein hydrolysates Thermoseparating polymer Viscera Aqueous two-phase systems Alkaline proteases |
author |
Ketnawa, Sunantha Benjakul, Soottawat Martínez Álvarez, Óscar Rawdkuen, Saroat |
author_facet |
Ketnawa, Sunantha Benjakul, Soottawat Martínez Álvarez, Óscar Rawdkuen, Saroat |
author_sort |
Ketnawa, Sunantha |
title |
Thermoseparating aqueous two phase system for the separation of alkaline proteases from fish viscera |
title_short |
Thermoseparating aqueous two phase system for the separation of alkaline proteases from fish viscera |
title_full |
Thermoseparating aqueous two phase system for the separation of alkaline proteases from fish viscera |
title_fullStr |
Thermoseparating aqueous two phase system for the separation of alkaline proteases from fish viscera |
title_full_unstemmed |
Thermoseparating aqueous two phase system for the separation of alkaline proteases from fish viscera |
title_sort |
thermoseparating aqueous two phase system for the separation of alkaline proteases from fish viscera |
publisher |
Marcel Dekker |
publishDate |
2014 |
url |
http://hdl.handle.net/10261/112395 |
work_keys_str_mv |
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1777670506796810240 |