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中国精品科技期刊2020
吴晓龙,王钰堡,刘容旭,等. 高静压酶解法制备猪小肠肝素及其稳定性研究[J]. 食品工业科技,2024,45(16):209−219. doi: 10.13386/j.issn1002-0306.2023090028.
引用本文: 吴晓龙,王钰堡,刘容旭,等. 高静压酶解法制备猪小肠肝素及其稳定性研究[J]. 食品工业科技,2024,45(16):209−219. doi: 10.13386/j.issn1002-0306.2023090028.
WU Xiaolong, WANG Yubao, LIU Rongxu, et al. Study of Preparation of Porcine Small Intestinal Heparin by High Hydrostatic Pressure Enzymatic Hydrolysis and Its Stability[J]. Science and Technology of Food Industry, 2024, 45(16): 209−219. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023090028.
Citation: WU Xiaolong, WANG Yubao, LIU Rongxu, et al. Study of Preparation of Porcine Small Intestinal Heparin by High Hydrostatic Pressure Enzymatic Hydrolysis and Its Stability[J]. Science and Technology of Food Industry, 2024, 45(16): 209−219. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023090028.

高静压酶解法制备猪小肠肝素及其稳定性研究

Study of Preparation of Porcine Small Intestinal Heparin by High Hydrostatic Pressure Enzymatic Hydrolysis and Its Stability

  • 摘要: 本研究以猪小肠粘膜为原料,通过响应面试验优化高静压酶解法提取肝素(Heparin,Hep)的最佳工艺,研究大孔树脂纯化Hep吸附条件,并探讨Hep的稳定性(pH、温度、光照)。结果表明:最佳酶解工艺为选取碱性蛋白酶,时间3.1 h,温度55 ℃,pH9.5,压力100 MPa,酶底物比9%,盐浓度2.0%,测得Hep实际平均浓度为11.56±0.12 U/mL;利用D958树脂纯化Hep,上样流速1.5 BV/h,洗脱液浓度4 mol/L NaCl,洗脱流速1.5 BV/h,纯化后Hep纯度可达68.75%,即达到电泳纯;在稳定性研究中,Hep在pH为7.5时稳定性最好,温度在40 ℃时具有较好稳定性,光照对Hep稳定性没有影响。本研究可大幅度提升Hep的提取效果,为猪小肠高值化加工利用提供理论和技术支持。

     

    Abstract: In this study, the mucosa of porcine small intestine was used as the raw material to optimize the optimal process for the extraction of heparin (Hep) by high hydrostatic pressure enzymatic hydrolysis by response surface test, to study the adsorption conditions of Hep purified by macroporous resin, and to explore the stability of Hep (pH, temperature, and light). The results showed that the optimal enzymatic process was alkaline protease, 3.1 h, 55 ℃, pH9.5, pressure 100 MPa, enzyme addition 9%, salt concentration 2.0%, the actual average concentration of Hep was measured to be 11.56±0.12 U/mL. The purification of Hep was carried out by using the D958 resin, with the up-sampling flow rate of 1.5 BV/h, the concentration of the eluent of 4 mol/L NaCl, and the elution flow rate of 1.5 BV/h, the purity of Hep could reach 68.75% after purification, i.e. electrophoretic purity was achieved. In the stability study, Hep had the best stability at pH7.5, better stability at 40 ℃, and light had no effect on the stability of Hep. This study can greatly improve the extraction effect of Hep and provide theoretical and technical support for the high value processing and utilization of pig small intestine.

     

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