An Integrated Method Combining Aqueous Two-phase Extraction and Boric Acid Affinity Adsorption for Separation and Purification of Horseradish Peroxidase from Horseradish Roots

doi:10.7506/spkx1002-6630-20201209-105

FOOD SCIENCE ›› 2022, Vol. 43 ›› Issue (2): 55-61.doi: 10.7506/spkx1002-6630-20201209-105

• Bioengineering • Previous Articles Next Articles

An Integrated Method Combining Aqueous Two-phase Extraction and Boric Acid Affinity Adsorption for Separation and Purification of Horseradish Peroxidase from Horseradish Roots

WU Yajiao, XING Youyuan, HAN Juan, WU Jiacong, LI Yuanyuan, WANG Yun

(1. School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang 212003, China; 2. School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212003, China; 3. Jingjiang College, Jiangsu University, Zhenjiang 212003, China)

Online:2022-01-25 Published:2022-01-29

Abstract

Abstract: The aim of the present work was to develop a novel method integrating two-step aqueous two-phase extraction and affinity adsorption for the separation and purification of horseradish peroxidase (HRP) from horseradish roots. Two-step aqueous two-phase extraction was used to remove sugars and pigments, and then “multivalent” phenylboronic acid affinity magnetic graphene composite (d-PBA-GO@Fe3O4@PEI) was to selectively isolate HRP from impurity proteins. The liquid-liquid equilibrium behaviors of 17R4-(NH4)2SO4, K3C6H5O7 or K2HPO4 aqueous two-phase systems (ATPSs) and the partition behaviors of nine model compounds in these R4-salt ATPSs were investigated, indicating that the optimum ATPS was 8% 17R4-13.2% (NH4)2SO4. The successful isolation of HRP from horseradish roots was achieved by the integrated method without negative effect on the structure of HRP, and the specific activity and purification factor of HRP were 28.93 U/mg and 1.92, respectively. The method has excellent separation and purification performance and thus holds great promise for application in the industrial production of plant glycoproteins.

Key words: horseradish peroxidase; purification; aqueous two-phase system; affinity adsorption; phenylboronic acid

CLC Number:

TS201.2

WU Yajiao, XING Youyuan, HAN Juan, WU Jiacong, LI Yuanyuan, WANG Yun. An Integrated Method Combining Aqueous Two-phase Extraction and Boric Acid Affinity Adsorption for Separation and Purification of Horseradish Peroxidase from Horseradish Roots[J]. FOOD SCIENCE, 2022, 43(2): 55-61.

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An Integrated Method Combining Aqueous Two-phase Extraction and Boric Acid Affinity Adsorption for Separation and Purification of Horseradish Peroxidase from Horseradish Roots

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