TY - JOUR
T1 - Fast separation and characterization of water-soluble proteins in wheat grains by reversed-phase ultra performance liquid chromatography (RP-UPLC)
AU - Yu, Zitong
AU - Han, Caixia
AU - Wang, Shu
AU - Lv, Dongwen
AU - Chen, Guanxing
AU - Li, Xiaohui
AU - Jiang, Guo Liang
AU - Yan, Yueming
N1 - Funding Information:
This research was financially supported in part by the grants from the National Natural Science Foundation of China ( 30830072 , 31271703, 31101145 ), the Chinese Ministry of Science and Technology (2009CB118303) and the National Key Project for Transgenic Crops of China (2011ZX08009-003-004 and 2011ZX08002-004).
PY - 2013/5
Y1 - 2013/5
N2 - Water-soluble proteins account for about 10% of total grain proteins in wheat (Triticum aestivum L.) and have specific functions in plant growth and development. In this study, a reversed-phase ultra performance liquid chromatography (RP-UPLC) method was trialed and the experimental conditions were optimized for rapidly separating and characterizing water-soluble proteins in wheat grains, and a comparative analysis with traditional RP-HPLC was performed. Under optimized separation conditions, fast, high resolution and reproducible RP-UPLC separation for water-soluble proteins could be obtained by gradually increasing eluting gradient from 21% to 47% in 30 min at 0.6 ml/min and 60 °C. Using this method, separation and characterization of water-soluble protein in wheat grains could be completed in less than 20 min for one sample, and the resolution and efficiency were significantly higher than those obtained with RP-HPLC. In addition, RP-UPLC consumed smaller amounts of samples and reagents as well. The optimized RP-UPLC could be used as an effective and alternative method for rapid separation and characterization of water-soluble proteins in wheat cultivar and germplasm evaluation, genetic and biochemical studies on grain proteins, and environmental influence analysis of water-soluble proteins.
AB - Water-soluble proteins account for about 10% of total grain proteins in wheat (Triticum aestivum L.) and have specific functions in plant growth and development. In this study, a reversed-phase ultra performance liquid chromatography (RP-UPLC) method was trialed and the experimental conditions were optimized for rapidly separating and characterizing water-soluble proteins in wheat grains, and a comparative analysis with traditional RP-HPLC was performed. Under optimized separation conditions, fast, high resolution and reproducible RP-UPLC separation for water-soluble proteins could be obtained by gradually increasing eluting gradient from 21% to 47% in 30 min at 0.6 ml/min and 60 °C. Using this method, separation and characterization of water-soluble protein in wheat grains could be completed in less than 20 min for one sample, and the resolution and efficiency were significantly higher than those obtained with RP-HPLC. In addition, RP-UPLC consumed smaller amounts of samples and reagents as well. The optimized RP-UPLC could be used as an effective and alternative method for rapid separation and characterization of water-soluble proteins in wheat cultivar and germplasm evaluation, genetic and biochemical studies on grain proteins, and environmental influence analysis of water-soluble proteins.
KW - Triticum aestivum L.
KW - Water-soluble proteins
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U2 - 10.1016/j.jcs.2012.11.008
DO - 10.1016/j.jcs.2012.11.008
M3 - Article
AN - SCOPUS:84877123626
SN - 0733-5210
VL - 57
SP - 288
EP - 294
JO - Journal of Cereal Science
JF - Journal of Cereal Science
IS - 3
ER -