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Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins
Published on: October 4, 2017
Synergistic Modification of Soybean Protein Isolate by Phosphorylation and Glycosylation for Enhanced Astaxanthin
Hua Jin1, Wenkang Li1, Wanze Zhang1
1College of Arts and Sciences, Northeast Agricultural University, Harbin 150030, China.
Abstract:
In this study, a novel combination strategy of sodium trimetaphosphate (STMP) phosphorylation and dextran (DX) glycosylation was employed to modify soy protein isolate (SPI). The phosphorylated protein-dextran conjugate (TSPI-DX) was successfully prepared and then was used as an emulsifier to prepare the astaxanthin emulsion, with the aim to enhance the emulsion delivery performance. Structural analysis revealed that phosphorylation and glycosylation altered the microenvironment of the side chains, leading to changes in protein secondary structure, which consequently loosened the protein architecture and enhanced molecular flexibility. The functional properties of TSPI-DX, including its solubility, emulsifying activity (EAI) and emulsifying stability (ESI), were markedly enhanced. Furthermore, the concurrent modification through phosphorylation and the Maillard reaction yielded a synergistic effect, boosting the DPPH radical scavenging rate by 86.5% and increasing the ferric-ion reducing power nearly fourfold. The astaxanthin emulsion prepared by modified SPI also exhibited several advantages. The TSPI-DX emulsion exhibited a markedly smaller mean particle size and a larger absolute Zeta-potential value. Consequently, with the higher electrostatic repulsion and steric hindrance among the droplets, the astaxanthin emulsion prepared by TSPI-DX demonstrated superior encapsulation efficiency and stability across various conditions, including storage, oxidation, thermal, and pH challenges. Moreover, in vitro digestion experiments revealed that the modified SPI emulsion facilitated a higher extent of lipolysis and astaxanthin bioaccessibility. Therefore, this work proposes a novel strategy for constructing plant-protein emulsion systems with enhanced delivery and release capabilities.

