Extraction, purification and identification of antifreeze proteins from cold acclimated malting barley (Hordeum vulgare L.). (15th May 2015)

• Record Type: Journal Article
• Authors: Ding, Xiangli; Zhang, Hui; Chen, Haiying; Wang, Li; Qian, Haifeng; Qi, Xiguang
• Is Part Of: Food chemistry. Volume 175(2015)
• Page Start: 74
• Page End: 81
• Publication Date: 2015-05-15
• Subjects: AFPs antifreeze proteins -- BaAFPs barley antifreeze proteins -- THA thermal hysteresis activity; Barley (Hordeum vulgare L.) -- Cold acclimate -- Antifreeze proteins -- Infiltration–centrifugation -- Purification -- Identification; Disodium phosphate dodecahydrate (PubChem CID: 61456) -- Sodium dihydrogen phosphate (PubChem CID: 23673460) -- Sodium chloride (PubChem CID: 5234) -- Tromethamine (PubChem CID: 6503) -- Hydrochloric acid (PubChem CID: 313) -- Glycerol (PubChem CID: 753) -- Bromophenol blue (PubChem CID: 8272) -- 2-Mercaptoethanol (PubChem CID: 1567) -- Ammonium sulphate (PubChem CID: 6097028); Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2014.11.027
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: LF-NMRI was used to monitor water migration of barley grain during infiltration. Infiltration–centrifugation was demonstrated to be targeted in BaAFPs extraction. BaAFPs was demonstrated to be accumulate after cold-acclimation. BaAFP-I at an electrophoresis level was obtained after a series of purification. BaAFP-I was identified to be an homology of alpha-amylase inhibitor BDAI-1. Abstract: Antifreeze proteins from cold-acclimated malting barley were extracted by infiltration–centrifugation. The infiltration time was optimised, and its extraction effect was evaluated. The effect of cold acclimation on the accumulation of barley antifreeze proteins (BaAFPs) was assessed by comparing the thermal hysteresis activities (THA) of proteins extracted from both cold acclimated and non-cold acclimated barley grain. Ultra-filtration, ammonium precipitation and column chromatography were used successively to purify the BaAFPs, and MALDI-TOF-MS/MS was used for protein identification. The results showed that infiltration–centrifugation was more targeted than the traditional method, and 10 h was the optimal infiltration time. THA was observed only after cold acclimation implied that AFPs only began to accumulate after cold acclimation. After purification, BaAFP-I was obtained at an electrophoresis level and its THA was 1.04 °C (18.0 mg ml −1 ). The mass fingerprinting and sequencing results indicated the homology of the BaAFP-I to alpha-amylase inhibitor BDAI-1 ( Hordeum vulgare ).