冰乙酸和胃蛋白酶依次提取新鲜林蛙皮胶原蛋白

采用冰乙酸和胃蛋白酶依次对新鲜林蛙皮胶原蛋白进行提取.在冰乙酸提取过程中,首先通过控制单一因素变量对料液比、pH值、冰乙酸浓度、酸解时间和温度等5个因素对新鲜林蛙皮胶原蛋白提取率的影响进行研究,并采用正交实验考察林蛙皮胶原蛋白酸解提取的最佳工艺;其次,在冰乙酸提取的最优条件下,利用胃蛋白酶将酸解后的残余物进行再次酶解提取.实验结果表明,最佳酸解的工艺条件是:料液比为1∶30,pH为7.5,酸解时间为48 h,酸解温度为4℃,冰乙酸浓度为0.5 mol/L.在此工艺条件下,胶原蛋白的提取率为11.85％,胃蛋白酶对蛙皮残渣的提取率为5.25％,所得胶原蛋白的纯度为64.70％.     

壳聚糖固定胃蛋白酶的研究

以壳聚糖为载体、戊二醛为交联剂将胃蛋白酶固定化．戊二醛质量分数为5％，载体处理温度为30℃，胃蛋白酶与壳聚糖用量比为O．Ol，pH=4．O，固定12h以上，酶活性回收率达65％～70％。固定化酶最适温度为70℃，比游离酶提高了20℃；固定化酶最适pH值为3．O，而游离酶为2．O；固定化酶的表观米氏常数Km=3．32g／L，而游离酶Km=1．5lg／L。固定化酶重复使用3次时的酶活性回收率分别为73．9％，58．4％。42．6％。2个多月重复使用，活力未见明显下降。     

Effects of pepsin hydrolysis on the soy β‐conglycinin aggregates formed by heat treatment at different pH

The effects of pepsin hydrolysis on the β‐conglycinin aggregates formed by heat treatment at different pH were investigated. Results showed that fibrils were still observed, whereas the random aggregates were easily to be digested in the simulated gastric fluid. Electrophoresis and molecular weight analysis indicated that large aggregates still existed after pepsin treatment for fibrils. Hydrolysis resulted in changes in the apparent viscosity (ηapp) of 6% fibril solutions. The ηapp at the shear rate range (0–30 s^?1) increased in the order of fibrils < fibrils with pepsin for 60 min < fibrils with pepsin for 30 min. Smaller peptide/fibril fragments were generated, and additional aggregates were reformed during the hydrolysis process, as evidenced by thioflavin T and atomic force microscopy images. The native β‐conglycinin hydrolysates comprised a mixture of polypeptides enriched in about 47 kDa. These findings would provide valuable information about effects of enzymatic hydrolysis on plant oligometric globulin aggregates.     

Hydrolytic degree and antioxidant activity of purified casein characterised by different haplotypes (αs1-, β- and k-casein) after enzymatic hydrolysis with pepsin and enzymatic extract from Pleurotus eryngii

The aim of this study was to estimate the hydrolytic degree and antioxidant activity of purified casein characterised by different haplotypes (α_s1-, β- and k-casein) after in vitro digestion with two different enzymatic systems: pepsin from porcine gastric mucosa (EP) and a crude enzymatic extract from the edible mushroom Pleurotus eryngii. The used enzymes showed a different mode of casein catalysis with a consequent production of peptides of different antioxidant activity. The CN haplotype significantly influenced peptides production; in fact, the amino acid substitutions caused by genetic polymorphisms at the α_s1-, β- and k-CN loci influenced the sites of enzymatic cleavage and therefore the produced peptides. The above is evidenced by the different antioxidant activity found in the hydrolysates depending on the used enzymatic system, the CN haplotype, and the CN haplotype × enzymatic treatment interaction. The findings of this study are a perspective for the production of specific foods that exert a biological effect in addition to the nutritional one.     

双水相分离提取胃蛋白酶和琼脂糖的初步研究

以聚乙二醇/硫酸钠双水相体系分离提取琼脂糖和胃蛋白酶.结果表明:琼脂糖的回收率高于90%,胃蛋白酶的回收率约为50%,两者最适宜的分离条件为:pH值为3,Na2SO4浓度为0.4 g·mL-1,PEG10000的浓度为0.002 mol·L-1.     

水解动物蛋白(HAP)的酶法制备及应用

报道了畜禽肉及鱼用酶解法制备ＨＡＰ的最适条件。当用双酶水解时，先用胰蛋白酶后用胃蛋白酶，水解效果最好。酶—酸联合水解，胶体磨处理及热变性，都能提高蛋白质水解率。ＨＡＰ有广泛应用前景。     

Pepsin-induced changes in the size and molecular weight distribution of bovine casein during enzymatic hydrolysis

Bovine casein was digested with pepsin at pH 2.0 in a batch-stirred tank reactor. To investigate the effect of peptic digestion on the aggregate size and molecular weight distribution of bovine casein, the resulting hydrolysates were examined by size-exclusion chromatography coupled with multiangle laser light scattering and dynamic light scattering. Casein was resolved by size-exclusion chromatography into 2 major peaks corresponding to aggregates and monomers, both of which showed a continuous decrease as hydrolysis proceeded. However, the ratio of aggregates to monomers was maintained at almost 1 (2:2.5) during the initial 30-min hydrolysis, indicating that the caseins in solution were in a type of equilibrium between aggregates and monomers. Upon peptic hydrolysis, casein aggregates increased in size and molecular weight, and exhibited a decrease in intermolecular repulsion. This finding was confirmed by dynamic light scattering measurements, which traced the changes in the hydrodynamic radii and light scattering intensities of casein hydrolysates. In addition, the release kinetics of peptide fractions with different molecular weights was also examined. It was concluded that the increase in hydrophobic attraction and the reduction in intermicellar repulsion might promote the growth in aggregate size of bovine casein during the limited hydrolysis.     

猪皮胶原蛋白水解物体外抗氧化作用模式初探

通过测定不同浓度不同水解时间的猪皮胶原蛋白水解物的还原能力、清除自由基能力、抗脂质氧化能力、金属离子鳌合能力来探讨其体外抗氧化作用机制。结果表明:在一定水解范围内,猪皮胶原蛋白水解物的抗氧化活性随着底物浓度的增加而增加,且水解物的还原能力、自由基清除能力、抗脂质氧化能力及Cu2+和Fe2+鳌合能力均随水解时间的延长而增大;底物浓度为4%、水解6 h的猪皮胶原蛋白水解物与其他水解条件下的水解物相比,具有较高的抗脂质氧化能力和较强的清除自由基能力,且具有丁基羟基甲苯(butylated hydroxytolu-ene,BHT)、抗坏血酸(ascorbic acid,Vc)等抗氧化剂所不具备的金属鳌合能力;4%的猪皮胶原蛋白6h水解物与0.02%Vc的还原能力相近。猪皮胶原蛋白水解物具有较高的抗氧化活性,可以作为氢供体、自由基稳定剂和金属离子螯合剂来抑制脂肪氧化。     

Application of ultrafiltration to the preparation of defined hydrolysates of bovine haemoglobin

Many uses of protein hydrolysates have been developed and applied to areas such as nutritional therapy, culture media, and the isolation of biologically active peptides. All these applications need carefully controlled and characterized hydrolysates. In order to produce such a type of hydrolysate, it is possible to use haemoglobin which is a very well defined and constant protein source. Enzymic hydrolysis of haemoglobin by pepsin was carried out at pilot-plant scale in an ultrafiltration reactor with mineral membranes. The object was to obtain a reproducible, decolorized, salt-free enzymic hydrolysate. Two types of membranes were tested having 10000 dalton (M5 type) and 20000 dalton (M4 type) cut-offs. Little significant difference was observed in the final products when both types of membranes were used. Reproducibility of hydrolysates was verified by amino acid analysis and gel filtration chromatography. The haemoglobin hydrolysates produced contained more than 90% protein and are especially suitable for fine applications.     

酶解花生蛋白影响Fe2+吸收利用的体外实验研究

以花生蛋白经模拟胃液和胰液消化所得的消化物为研究对象,研究酶解花生蛋白对Fe2 生物效价的影响。结果表明,随Fe2 浓度的增加,花生蛋白模拟消化物与Fe2 的结合量下降,且随着时间的延长而降低;酶解花生蛋白能保护Fe2 不易被氧化为Fe3 ;Ca2 的加入能降低酶解花生蛋白与Fe2 的结合量。