不同提取方法的马骨髓蛋白结构及活性比较研究

Comparative Study on the Structure and Activity of Horse Bone Marrow Protein Extracted by Different Methods

研究不同提取方法对马骨髓蛋白提取率、结构及抗菌、抗氧化活性的影响,探讨其食用和药用价值。以蛋白含量、提取率、分子量及抗氧化活性为考察指标筛选12种提取方法,采用UV、SDS-PAGE、FT-IR、SEM-EDX进行结构表征;通过体外抗菌和抗氧化活性实验评价其生物活性。经综合考察得出水、0.25%NaCl(质量分数)及生理盐水提取所得马骨髓蛋白指标较高,蛋白含量分别为41.91、39.93、37.82 mg/mL,提取率为19.26%、14.86%、23.22%。UV图谱显示不同方法制备的马骨髓蛋白在270 nm附近出现最大吸收峰,SDS-PAGE显示蛋白分子量主要集中在66 ku附近、且出现相应的多肽条带,FT-IR图谱显示其在4000~400 cm^(-1)区域均出现典型的蛋白吸收峰,SEM-EDX显示其蛋白由球状,条状和不规则颗粒构成。生物活性试验结果表明以上三种方法所得的马骨髓蛋白具有较好的抑菌及抗氧化作用,对金黄色葡萄球菌的抑菌圈直径分别为12.0、16.0、11.0 mm;清除DPPH及ABTS自由基的IC_(50)分别为0.32、0.29、0.31 mg/mL及0.26、0.15、0.20 mg/mL。研究结果可为马肉副产物骨类资源的进一步开发利用提供科学依据。

The effects of different extraction methods on extraction rate,structure,antibacterial activity and antioxidant activity of horse bone marrow protein were studied,and its edible and medicinal values were examined.Twelve extraction methods were screened through using protein content,extraction rate,molecular weight and antioxidant activity as the indicators,and structural characterizations were performed by UV,SDS-PAGE,FT-IR,and SEM-EDX analysis.The biological properties of the obtained proteins were evaluated by in vitro antibacterial and antioxidant activities.After comprehensive investigations,it was found that the index scores of the horse bone marrow proteins extracted with water,0.25%NaCl,and normal saline were higher,with their corresponding protein contents being 41.91,39.93 and 37.82 mg/mL,and extracted rates being 19.26%,14.86%and 23.22%,respectively.UV spectra showed that the horse bone marrow proteins prepared by different methods had the maximum absorption peak around 270 nm.SDS-PAGE revealed that the molecular weights of the obtained proteins were mainly around 66 ku,with the presence of corresponding polypeptide bands.FT-IR spectra showed typical protein absorption signals in the region of 4000~400 cm^(-1).SEM-EDX analysis showed that the obtained proteins were composed of spherical,striped and irregular particles.The results of biological activity experiments demonstrated that the horse bone marrow proteins prepared by the three methods exhibited sound antibacterial and antioxidant effects,and the zone diameters of inhibition against Staphylococcus aureus were 12.0,16.0 and 11.0 mm,respectively.The IC_(50) values of DPPH and ABTS free radicals scavenging activities were 0.32,0.29 and 0.31 mg/mL,and 0.26,0.15 and 0.20 mg/mL,respectively.The research results can provide a scientific basis for further development and utilization of horse meat processing by-products,bone resources.