乳源性生物活性肽对新生IUGR仔猪肝脏抗氧化功能的影响

本试验选择苏太初生IUGR仔猪10窝共20头,配对分为4组:胰岛素组(I组)、牛乳组(M组)、酪蛋白酶解产物组(CH组)和酪蛋白组(C组),饲喂3d后宰杀。另选5只初生IUGR仔猪立即宰杀,作为0d对照组(A组)。M组喂牛乳,I组在牛乳中添加2.5mg/L胰岛素;C组用1.5%酪蛋白水溶液取代10%(V/V)的牛乳,CH组用1.5%酪蛋白酶解液取代10%(V/V)的牛乳。宰杀后分析肝脏和血浆中SOD活性和MDA含量以及血浆中GPT和GOT水平。试验结果显示:酪蛋白酶解物可显著降低肝脏和血浆MDA含量,提高SOD活性(P<0.05),胰岛素可显著降低血浆中MDA含量(P<0.05);3日龄仔猪GPT和GOT均高于初生仔猪,其中I组、C组和CH组GPT均显著高于M组(P<0.05)。结果提示:胰岛素和酪蛋白酶解物可提高IUGR仔猪肝脏的抗氧化功能,促进肝细胞的生长发育。     

仔猪胃肠道食糜和血液中酪啡肽的检测及阿片样活性分析

将24头仔公猪分为对照组(正常哺乳28 d断奶)、试验I组(添喂β-酪啡肽-7)和试验Ⅱ组(添喂酪蛋白酶解物)。试验组于21 d断奶,再连续添喂酪蛋白胃蛋白酶解液或β-酪啡肽-7 10 d,每天2次(共20mL),至32 d,3组动物各随机取6头同时剖杀,取血、胃食糜和空肠食糜。通过所建立的RP-HPLC方法,在胃食糜和空肠食糜中检测到β-酪啡肽-5、β-酪啡肽-7样物质。对照组、酪啡肽-7组和酪蛋白水解液组,β-酪啡肽-7的平均含量分别为21.12±2.38,23.58±7.49、21.81±9.93μg/g胃食糜和16.99±3.48、19.08±2.85、21.36±4.85μg/g肠食糜;胃食糜中-βCM-7的含量高于空肠食糜,试验组均高于对照组。在各组仔猪的血液中未检测到酪啡肽样物质。肠食糜的细胞生物学试验结果表明:与喂仔猪料的对照组比较,试验I组和试验Ⅱ组对细胞腺苷酸环化酶的活性均具有不同程度的抑制作用,抑制率分别为27.9%和89.0%,抑制作用均能被纳洛酮反转,逆转率分别是43.2%和89.07%。提示在21日龄断奶仔猪饲料中添加酪啡肽或酪蛋白酶解物,其肠道中有酪啡肽释放,阿片总活性升高。     

Inter‐related effects of dietary fat and protein level on growth performance in rabbits

Isoenergetic substitution of dietary corn oil for dietary carbohydrates enhances growth in rabbits. It was hypothesized that identical amounts of metabolizable energy in the form of corn oil are more effective than those of carbohydrates in reducing protein catabolism and thus sparing it for growth, which would imply that the fat effect is greater on a marginal than normal protein diet. Young growing rabbits were fed semi‐purified diets either relatively high (21.6 energy % protein) or low in casein (13.0 energy % protein) to which extra corn oil (21.1 instead of 5.3 energy %) was added at the expense of an isoenergetic amount of corn starch and dextrose. The addition of corn oil to the diet with 21.6 energy % protein indeed increased weight gain, but the addition to the diet with 13.0 energy % protein left weight gain unchanged. These results refute our hypothesis, because the low‐protein intake was not limiting growth. The enrichment of the high‐protein diet with extra corn oil did not affect nitrogen retention, whereas urinary nitrogen excretion was increased. These observations also are at variance with the idea that additional fat would spare nitrogen for growth.     

Comparison of methods for assay of fecal proteolytic activity

Fecal proteolytic activity (FPA) in ten normal dogs was readily detected either calorimetrically using azocasein substrate or by radial enzyme diffusion into agar gels containing casein substrate. Daily FPA ranged from 17 to 207 azocasein units/g (ACUIG) or 4 to 18 mm of casein hydrolysis, while mean 3-day FPA ranged from 58 to 10 1 ACUIG or 7 to 15 mm. Studies of proteolytic activity remaining after treatment of fecal extracts with a specific trypsin inhibitor indicated that trypsin accounted for 0% to 71% of proteolytic activity. Proteolytic activity decreased steadily in fecal specimens stored at room temperature or above, but there was only slight loss in activity during storage for up to 5 days at 4 degrees C. Proteolytic activity was unaffected by repeated freezing and thawing and samples could be stored for long periods at -2 degrees C without noticeable loss of activity. It is concluded that assays of FPA using either azoprotein substrate or radial enzyme diffusion into agar gels containing casein substrate allow evaluation of exocrine pancreatic function in dogs, provided that several samples are tested. These methods are suitable for application in a variety of species.     

牛乳酪蛋白酶解物的分离纯化及抗菌肽乳基料的制备研究

【目的】分离纯化牛乳酪蛋白酶解物,检测酶解物及其分离组分的抑菌活性,并制备抗菌肽乳基料。【方法】从中性蛋白酶、碱性蛋白酶、胰蛋白酶及木瓜蛋白酶中筛选一种蛋白酶水解牛乳酪蛋白,将其酶解产物经大孔吸附树脂和凝胶过滤色谱分离、纯化后,以对大肠杆菌、沙门氏菌和金黄色葡萄球菌的抑菌圈直径、最小抑菌浓度(MIC)、最小杀菌浓度(MBC)为指标,分析酶解物和分离组分的抑菌活性。【结果】(1)木瓜蛋白酶水解牛乳酪蛋白4.5h,其酶解物对大肠杆菌、沙门氏菌和金黄色葡萄球菌均有较好的抑制作用。(2)大孔吸附树脂将牛乳酪蛋白酶解物分离成4个组分,其中以体积分数75%乙醇洗脱组分的抑菌作用较强。(3)凝胶过滤色谱将体积分数75%乙醇洗脱组分分离成4个色谱峰,其中A峰对大肠杆菌和沙门氏菌抑制作用较强,B峰对金黄色葡萄球菌抑制作用较强,A峰和B峰经冷冻干燥后成功制备了2种抗菌肽乳基料。【结论】牛乳酪蛋白酶解物经大孔吸附树脂和凝胶过滤色谱分离、纯化后,其分离组分的抑菌活性逐步增强,能够用于抗菌肽乳基料的制备。     

利用毛细管电泳测定牛乳和山羊乳混合乳的蛋白质

采用毛细管区带电泳方法,选择聚丙烯酰胺涂层毛细管对混合乳中的蛋白进行图谱的研究,确定混合乳的定性定量检测方法。该方法对牛乳和山羊乳混合乳中的蛋白进行了很好的分离,确定了牛乳αs1-CN、αs0-CN、κ-CN、β-casein A1和山羊乳的β-CN、β1-casein作为定性检测的酪蛋白,测得了混合乳中不同酪蛋白峰面积的比值与混合比例呈很好的线性关系,相关系数均大于0.997,可以作为定量检测混合乳的指标。本法简便、快速、准确,为乳及乳制品质量的监控提供了一种新的手段,可用于乳的质量监控。     

Degradation of two soluble proteins – casein and egg protein by a macro in vitro method

Degradation of casein and egg protein was studied with whole rumen contents (RC) in a macro in vitro system to elucidate previous findings of initial rapid disappearance of soluble proteins in vitro. Five to 7.5 kg of RC from a dry and/or a lactating cow were incubated with buffer and casein or egg protein for 180 min with frequent sampling. Degradation was measured as loss of trichloroacetic acid precipitable N (TCA‐N) from the inocula. Normal (39 °C) and low (2 °C) temperature incubations were examined in Exp. 1, using 1 g of TCA‐N from casein. Four levels of casein (0–12 g TCA‐N) in Exp. 2 and four levels of egg albumin (0–24 g TCA‐N) in Exp. 3 were fermented at 39 °C. Initial recovery of casein TCA‐N was 106% at 2 °C and 56% at 39 °C (Exp. 1). Casein (TCA‐N) recovered initially increased in Exp. 2 from 21% at 3 g to 86% at 12 g TCA‐N, while absolute loss remained relatively constant at 358 mg TCA‐N/kg RC (SD = 47). Fractional degradation rate was highest (0.03/min) at the intermediate dosage level. In the absence of rumen fluid (Exp. 4), no casein was lost. Initial egg protein recovery was on average 103% (Exp. 3). Recovery seemed unaffected by dosage level, and absolute degradation rate was relatively constant over time and increased with dosage level (p < 0.001) from 1.48 to 2.95 mg TCA‐N/(kg RC × min). Maximum degradation rate [mg TCA‐N/(kg RC × min)] and affinity constant (mg TCA‐N/kg RC) were estimated at 261 and 1650, respectively. It is concluded that a surprisingly constant amount of casein disappears immediately from warm rumen fluid and that this does not occur either with chilled RC, in the absence of rumen fluid, or when replaced with egg protein. The mechanisms for this disappearance are yet to be discovered.     

牛乳酪蛋白水解物的特性分析

牛乳酪蛋白经胰蛋白酶水解，对水解物进行氨基氮含量、水解度（DH）和游离氨基酸含量等测定，并通过SDS－PAGE电泳及Sephadex-G50凝胶层析，分析了相对分子质量的分布。结果表明，在水解条件为底物质量浓度50g/ml、pH7．4、温度45℃、酶量（m)：底物量（m）＝1：1000时，DH随时间延长而增大，水解前期DH增长迅速，3h后增长缓慢，6．0h的DH为20．48％。水解物中必需氨基酸含量约占游离氨基酸总量的73％－80％，此含量随水解时间的延长略有降低；水解物相对分子质量随DH的升高有所减小，经2－3h水解的产物，相对分子质量分布主要集中于2500－3400范围。     

微小毛霉凝乳酶干酪素的制备及应用性能研究

以新鲜牛奶为原料,采用自制微小毛霉Mucor Pusilius凝乳酶作为脱脂牛奶凝固剂,经过离心脱脂、巴氏杀菌、凝乳、热烫、离心脱乳清、水洗、干燥、粉碎过筛等步骤获得凝乳酶干酪素,考察工艺参数对干酪素得率及应用性能的影响.实验结果表明,凝乳酶添加量、凝乳pH值、热烫温度对凝乳酶干酪素得率无显著影响,热烫温度对产品的融化...     

Standardized ileal digestibility of proteins and amino acids in sesame expeller and soya bean meal in weaning piglets

Apparent ileal digestibility (AID) of diets containing sesame expeller (SE) and soya bean meal (SBM) was determined using 15 piglets (Genetiporc^®), weaned at 17 ± 0.4 days with average body weight of 6.4 ± 0.7 kg (Fertilis 20 × G Performance, Genetiporc^®, PIC México, Querétaro, México). Piglets were randomly assigned to three treatments: (i) a reference diet with casein as the sole protein source; (ii) a mixed diet of casein–SE; and (iii) a mixed diet of casein–SBM. The chemical composition of SE and SBM was determined, and AID and standardized ileal digestibility (SID) of crude protein (CP) and amino acids (AAs) were determined for each protein source. SE contained greater quantities of ether extract, neutral detergent fibre, phytic acid, methionine and arginine than SBM. Lysine and proline contents and trypsin inhibitor activity were higher in SBM than in SE. The AID and SID of CP and AA (except for lysine and proline) were similar in SE and SBM. The AID of lysine and proline was higher in SBM than in SE (p < 0.05), and the SID of proline was higher in SE than in SBM (p < 0.05). These findings indicate that SE is an appropriate alternative protein source for early weaned pigs.