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<p>Opuntia milpa alta polysaccharides (OPS) possess diverse biological activities and exhibit great potential for application in pharmaceuticals and functional foods. However, conventional extraction methods usually yield OPS with low total sugar content and unsatisfactory bioactivity, and the effects of extraction protocols on the structural properties and bioactivities of OPS remain to be systematically elucidated. In this study, three extraction methods, including conventional hot water extraction (OPS-H), enzyme-assisted ultrasonic extraction (OPS-U), and deep eutectic solvent extraction (OPS-D), were systematically compared in terms of polysaccharide yield, total sugar content, structural characteristics, and enzyme inhibitory capacities. The results showed that OPS-D presented the highest extraction yield (39.46%) and total sugar content (70.39%), as well as a smaller particle size and lower molecular weight (75.45&#xa0;kDa) compared with OPS-H (131.23&#xa0;kDa). All OPS samples shared similar qualitative monosaccharide profiles but differed significantly in the molar ratios of individual monosaccharides, suggesting that the extraction protocols mainly regulated the molecular weight and monosaccharide proportions of OPS without altering the types of major monosaccharide components. Furthermore, OPS-D exhibited superior antioxidant activity and stronger inhibitory effects against α-glucosidase (IC₅₀ = 0.92&#xa0;mg/mL) and α-amylase (IC₅₀ = 0.8&#xa0;mg/mL). Enzyme kinetic analysis clarified the inhibition types of OPS against the target enzymes, and comparative fluorescence quenching assays covering all three OPS fractions were performed to elucidate the binding interaction mechanisms between OPS and the target enzymes. These findings indicate that deep eutectic solvent extraction is an efficient strategy for preparing OPS with enhanced in vitro antioxidant and carbohydrate-hydrolyzing enzyme inhibitory activities. This study provides experimental support for the high-value utilization of Opuntia milpa alta polysaccharides and the development of functional foods targeting carbohydrate digestive enzymes.</p>

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Effects of different extraction methods on the structure and enzyme inhibitory activity of Opuntia milpa alta polysaccharides

  • 期刊论文

  • 发表日期：

    2026年7月21日

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  • Xinyi Ma,
  • Xin Wang,
  • Xinhong Shi,
  • Xin Yang,
  • Haobin Zhu,
  • Yongqiang Ma

摘要

Opuntia milpa alta polysaccharides (OPS) possess diverse biological activities and exhibit great potential for application in pharmaceuticals and functional foods. However, conventional extraction methods usually yield OPS with low total sugar content and unsatisfactory bioactivity, and the effects of extraction protocols on the structural properties and bioactivities of OPS remain to be systematically elucidated. In this study, three extraction methods, including conventional hot water extraction (OPS-H), enzyme-assisted ultrasonic extraction (OPS-U), and deep eutectic solvent extraction (OPS-D), were systematically compared in terms of polysaccharide yield, total sugar content, structural characteristics, and enzyme inhibitory capacities. The results showed that OPS-D presented the highest extraction yield (39.46%) and total sugar content (70.39%), as well as a smaller particle size and lower molecular weight (75.45 kDa) compared with OPS-H (131.23 kDa). All OPS samples shared similar qualitative monosaccharide profiles but differed significantly in the molar ratios of individual monosaccharides, suggesting that the extraction protocols mainly regulated the molecular weight and monosaccharide proportions of OPS without altering the types of major monosaccharide components. Furthermore, OPS-D exhibited superior antioxidant activity and stronger inhibitory effects against α-glucosidase (IC₅₀ = 0.92 mg/mL) and α-amylase (IC₅₀ = 0.8 mg/mL). Enzyme kinetic analysis clarified the inhibition types of OPS against the target enzymes, and comparative fluorescence quenching assays covering all three OPS fractions were performed to elucidate the binding interaction mechanisms between OPS and the target enzymes. These findings indicate that deep eutectic solvent extraction is an efficient strategy for preparing OPS with enhanced in vitro antioxidant and carbohydrate-hydrolyzing enzyme inhibitory activities. This study provides experimental support for the high-value utilization of Opuntia milpa alta polysaccharides and the development of functional foods targeting carbohydrate digestive enzymes.