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Purification and Identification of Antifreeze Protein From Cold-Acclimated Oat (Avena sativa L.) and the Cryoprotective Activities in Ice Cream

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Abstract

Antifreeze proteins (AFPs) were extracted from cold-acclimated oat by vacuum infiltration–centrifugation. Ammonium precipitation, ion exchange, and size-exclusion chromatography were used successively to purify the oat AFPs (AsAFPs), and the proteins were identified using matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry. The results showed that an ammonium-sulfate saturation of 50–100 % was optimal for supernatant precipitation. After purification, AsAFP was found to have achieved an electrophoretic purity and its thermal-hysteresis activity was measured at 1.24 °C (15 mg ml−1). The mass fingerprinting and sequencing results indicated the homology of AsAFP and endochitinase. Recrystallization and melting resistance of ice cream were improved by AsAFP (0.1 %). Moreover, the addition of AsAFP (0.1 %) in ice cream increased the glass transition temperature (Tg) from −29.14 to −27.74 °C.

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Abbreviations

AFPs:

Antifreeze proteins

THA:

Thermal-hysteresis activity

DSC:

Differential scanning calorimeter

PBS:

Phosphate-buffered saline solution

DEAE:

Diethylaminoethyl cellulose

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

RIA:

Recrystallization inhibition activity

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Acknowledgments

Financial support from the National Natural Science Foundation of China (No. 31171637) is gratefully acknowledged.

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Authors and Affiliations

  1. State Key laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, 214122, Jiangsu Province, China

    Yanjie Zhang, Hui Zhang, Lilin Cheng, Li Wang, Haifeng Qian, Xiguang Qi & Chaoyang Song

  2. College of Tourism and Culinary Science, Yangzhou University, Yangzhou, 225127, China

    Xiangli Ding

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  1. Yanjie Zhang
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  2. Hui Zhang
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  3. Xiangli Ding
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  4. Lilin Cheng
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  5. Li Wang
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  6. Haifeng Qian
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  7. Xiguang Qi
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  8. Chaoyang Song
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Correspondence to Hui Zhang.

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Zhang, Y., Zhang, H., Ding, X. et al. Purification and Identification of Antifreeze Protein From Cold-Acclimated Oat (Avena sativa L.) and the Cryoprotective Activities in Ice Cream. Food Bioprocess Technol 9, 1746–1755 (2016). https://doi.org/10.1007/s11947-016-1750-x

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