Characterization of oat proteins and aggregates using asymmetric flow field-flow fractionation.
(2013) In Analytical and Bioanalytical Chemistry 405(21). p.6649-6655- Abstract
- The soluble proteins and protein aggregates in Belinda oats were characterized using asymmetric flow field-flow fractionation (AF4) coupled with online UV-vis spectroscopy and multiangle light-scattering detection (MALS). Fractions from the AF4 separation were collected and further characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The AF4 fractogram of the oat extracts revealed three peaks which were determined to be monomeric forms of soluble proteins, globulin aggregates, and β-glucan, respectively. The early eluting monomeric proteins ranged in molar mass (MM) between 5 and 90 kg/mol and in hydrodynamic diameter (D h) from 1.6 to 13 nm. The MM at peak maximum of the globulin aggregate peak was found... (More)
- The soluble proteins and protein aggregates in Belinda oats were characterized using asymmetric flow field-flow fractionation (AF4) coupled with online UV-vis spectroscopy and multiangle light-scattering detection (MALS). Fractions from the AF4 separation were collected and further characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The AF4 fractogram of the oat extracts revealed three peaks which were determined to be monomeric forms of soluble proteins, globulin aggregates, and β-glucan, respectively. The early eluting monomeric proteins ranged in molar mass (MM) between 5 and 90 kg/mol and in hydrodynamic diameter (D h) from 1.6 to 13 nm. The MM at peak maximum of the globulin aggregate peak was found to be ∼300 kg/mol and the D h was measured to be ∼20 nm. SDS-PAGE of the collected fraction across this peak revealed two bands with MM of 37 and 27 kg/mol which correspond to the α and β subunits of globulin indicating the elution of globulin aggregates. A third peak at long retention time was determined to be β-glucan through treatment of the oat extract with β-glucanase and by injection of β-glucan standards. The amount of soluble protein was measured to be 83.1 ± 2.3 wt.%, and the amount of albumin proteins was measured to be 17.6 ± 5.7 wt.% of the total protein in the oats. The results for Belinda oat extracts show that the AF4-MALS/UV platform is capable of characterizing the physicochemical properties such as MM and hydrodynamic size distribution of proteins and protein aggregates within a complicated food matrix environment and without the need to generate protein isolates. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/3956336
- author
- Runyon, Ray LU ; Nilsson, Lars LU ; Alftrén, Johan LU and Bergenståhl, Björn LU
- organization
- publishing date
- 2013
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Analytical and Bioanalytical Chemistry
- volume
- 405
- issue
- 21
- pages
- 6649 - 6655
- publisher
- Springer
- external identifiers
-
- wos:000322705600005
- pmid:23812878
- scopus:84881367384
- ISSN
- 1618-2642
- DOI
- 10.1007/s00216-013-7115-7
- language
- English
- LU publication?
- yes
- id
- 5417e380-a6dc-4eeb-90b4-f808f5a3adf7 (old id 3956336)
- date added to LUP
- 2016-04-01 09:49:26
- date last changed
- 2024-10-06 13:36:54
@article{5417e380-a6dc-4eeb-90b4-f808f5a3adf7, abstract = {{The soluble proteins and protein aggregates in Belinda oats were characterized using asymmetric flow field-flow fractionation (AF4) coupled with online UV-vis spectroscopy and multiangle light-scattering detection (MALS). Fractions from the AF4 separation were collected and further characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The AF4 fractogram of the oat extracts revealed three peaks which were determined to be monomeric forms of soluble proteins, globulin aggregates, and β-glucan, respectively. The early eluting monomeric proteins ranged in molar mass (MM) between 5 and 90 kg/mol and in hydrodynamic diameter (D h) from 1.6 to 13 nm. The MM at peak maximum of the globulin aggregate peak was found to be ∼300 kg/mol and the D h was measured to be ∼20 nm. SDS-PAGE of the collected fraction across this peak revealed two bands with MM of 37 and 27 kg/mol which correspond to the α and β subunits of globulin indicating the elution of globulin aggregates. A third peak at long retention time was determined to be β-glucan through treatment of the oat extract with β-glucanase and by injection of β-glucan standards. The amount of soluble protein was measured to be 83.1 ± 2.3 wt.%, and the amount of albumin proteins was measured to be 17.6 ± 5.7 wt.% of the total protein in the oats. The results for Belinda oat extracts show that the AF4-MALS/UV platform is capable of characterizing the physicochemical properties such as MM and hydrodynamic size distribution of proteins and protein aggregates within a complicated food matrix environment and without the need to generate protein isolates.}}, author = {{Runyon, Ray and Nilsson, Lars and Alftrén, Johan and Bergenståhl, Björn}}, issn = {{1618-2642}}, language = {{eng}}, number = {{21}}, pages = {{6649--6655}}, publisher = {{Springer}}, series = {{Analytical and Bioanalytical Chemistry}}, title = {{Characterization of oat proteins and aggregates using asymmetric flow field-flow fractionation.}}, url = {{http://dx.doi.org/10.1007/s00216-013-7115-7}}, doi = {{10.1007/s00216-013-7115-7}}, volume = {{405}}, year = {{2013}}, }