复酶降解羽毛蛋白工艺的研究

研究复酶对羽毛蛋白尤其是对角蛋白的降解效果。选择复酶酶解温度、酶解液pH、复酶添加量和酶解时间为因素水平,采用L9(34)正交试验设计,对酶解条件进行优选。确定最佳酶解参数:酶解液初pH为7,酶解温度为45℃,酶解时间为32h,复酶添加量为0.006g。结果表明:在试验确定的最佳酶解条件下,羽毛降解率达到了90.87%,酶解液可溶性蛋白的含量达到848.1mg/L,酶解上清液中蛋白分布在20.1～66.2ku,酶解羽毛蛋白体外消化率达到82.83%。     

大米蛋白胃蛋白酶酶解物体外抗氧化作用的研究

本试验采用正交设计,以羟自由基清除率为评定指标,用胃蛋白酶酶解大米蛋白,旨在确定其酶解物体外抗氧化作用最佳的酶解条件。试验结果表明:当酶解温度为37℃时,影响大米蛋白胃蛋白酶酶解物抗氧化效果的因素大小排序为:pH酶浓度底物浓度酶解时间;最佳酶解条件为:pH1.4、酶浓度5%、底物浓度8%、酶解120min,此条件下酶解物的羟自由基清除率为72.78%,多肽浓度为5.49mg/mL,水解度为15.62%。     

鸭空肠液中全消化酶与主要消化酶体外总消化能力的比较研究

本试验旨在研究北京鸭空肠液中淀粉酶,脂肪酶,胰蛋白酶和糜蛋白酶的总消化能力占所有酶总消化能力的比例,及其他未知酶发生变化后对测试结果的影响.采用单因素完全随机设计,根据肠液酶组成设3个处理,处理间4种主要酶的活性一致.处理1为鸭内源肠液(4种酶+其他酶),处理2为鸭肠液干粉剂+试剂酶(4种酶+其他酶),处理3为试剂酶(...     

大豆虾粉肽适宜酶解好条件的研究

为了筛选生产大豆虾粉肽的适宜蛋白酶及酶解条件,试验采用单因素试验设计,以水解度为检测指标,按酶浓度、底物浓度、pH值、酶解温度和酶解时间逐一分别对3种微生物发酵产生的酸性蛋白酶、碱性蛋白酶和中性蛋白酶水解脱皮豆粕和虾粉混合蛋白的适宜酶解条件进行筛选.试验结果表明:酶解体系中的酶浓度、底物浓度、pH值、酶解温度和酶解时间均显著影响3种蛋白酶对脱皮豆粕和虾粉混合蛋白的酶解效果.在3种蛋白酶中,酸性蛋白酶的酶解效果最好,其次是碱性蛋白酶,中性蛋白酶的酶解效果最差.酸性蛋白酶水解脱皮豆粕和虾粉混合蛋白的适宜条件为:酶浓度为2000 U/g,底物浓度为9%,pH值3.5,酶解时间为3.5 h,温度50℃.     

桑叶蛋白血管紧张素转换酶抑制肽的酶解制备

本研究旨在探讨桑叶蛋白血管紧张素转换酶(ACE)抑制肽的酶解制备方法,从木瓜蛋白酶、中性蛋白酶、胃蛋白酶、芽孢杆菌蛋白酶、碱性蛋白酶、胰蛋白酶6种蛋白酶中筛选出最佳蛋白酶,并运用单因素逐级优化法对酶解反应的底物浓度、加酶量、温度、pH、酶解时间进行参数优化.选取这6种常用蛋白酶,利用酶解法制备桑叶蛋白多肽,以ACE抑制率为主要指标,水解度为辅助指标,研究桑叶多肽对ACE抑制活性的影响.结果 表明,酶解效果最佳的酶为芽孢杆菌蛋白酶,最佳酶解参数为底物质量浓度20 g/L、加酶量7.5％、温度60℃、pH7.0和酶解时间50 min,此时酶解产物的ACE抑制率为81.51％,水解度为15.86％.     

超声波协同酶解法提取啤酒花黄酮类的工艺优化及其抗氧化试验

为探索超声波协同酶解法提取啤酒值花黄酮的最佳工艺,研究该方法下啤酒花黄酮的抗氧化活性,利用单因素法研究了不同酶量、酶解时间、酶解温度、酶解pH值对超声波协同酶解法提取啤酒花总黄酮含量的影响。结果表明,超声波协同酶解法明显提高啤酒花黄酮的提取率,最佳提取条件为:酶量为2 mg/g,酶解p H值为4,酶解温度30℃,酶解时间2 h,啤酒花总黄酮提取量55.7 mg/g,明显高于传统提取方法的总黄酮提取率38.5%,并对·OH和DPPH具有良好的清除效果,高于常用的抗氧化剂天然维生素C对两者的清除率。     

固定化生姜蛋白酶的制备及其性质研究

本试验以海藻酸钠为载体、戊二醛为交联剂,制备固定化生姜蛋白酶,对固定化条件进行优化并对固定化酶与游离酶的部分酶学性质进行研究。结果表明,固定化生姜蛋白酶的最佳条件是海藻酸钠浓度为2%,CaCl2的浓度为2%,戊二醛的浓度为3%,在此条件下制备的固定化酶活力为53.48 U/g。固定化酶的最适温度是60℃,最适pH值为6.0,与游离酶相比没有发生改变,但固定化酶与游离酶相比对温度和酸碱适应范围更加宽泛。     

酶法水解牛乳蛋白技术的研究

为获得易消化、低致敏的婴儿配方食品原料,采用胰蛋白酶水解牛乳蛋白,研究酶解过程中酶解时间、酶解温度、加酶量对乳蛋白水解度的影响。通过单因素试验设计方法,确定最佳酶解条件为:酶解时间40min、酶解温度40℃、加酶量1200U/g蛋白质。     

饲用α-半乳糖苷酶酶活稳定性的研究

在酶制剂生产加工和储存过程中,酶活稳定性是酶应用过程中最常遇到的实际问题。本文研究不同载体、高温高湿、金属离子、保存初始pH值、保护剂浓度和存储时间对体外α-半乳糖苷酶稳定性的影响。试验结果表明:①无机载体碳酸钙、元明粉和滑石粉对α-半乳糖苷酶活性影响很大,收率均为0%;有机载体中玉米皮粉效果最好,收率高达90.83%;其次是淀粉,收率为61.11%;稻壳粉收率最低,仅为44.72%。②湿度为17%,温度小于85℃,酶活损失率小于10%,说明该酶有较强的耐温能力,温度为90℃,酶活损失率达到16.2%,随着温度上升,酶活损失率开始增加,不利于酶活的保存。③Cu2+对该酶有抑制作用3,7℃恒温水浴4 h后酶活存留率仅为74%,Mn2+、Zn2+、Ca2+、Na+、K+和Mg2+对α-半乳糖苷酶有几近相同的保护作用,酶活存留率均大于92%。④该酶最适保存pH值为5.3;pH值为4.4时,酶活损失率达到30.1%,不利于酶的保存;pH值为5.6时,酶活损失率为3.77%,pH值在4.7~5.6之间,酶活损失率小于10%,有利于酶的保存。总之,过酸和过碱都不利于酶的保存。⑤1%NaCl、1%甘露醇、5%和7%的山梨醇都有杂菌产生,不利于酶的保存3;%~5%的NaCl酶活损失小于10%1,%和5%~7%的甘油酶活损失率小于5%,都有利于酶活保存。     

糙皮侧耳原生质体制备与再生条件的研究

以糙皮侧耳为材料,研究酶的组成、酶解时间、酶解温度、菌龄和渗透压稳定剂对糙皮侧耳原生质体制备率和再生率的影响.结果表明:静置培养7d的糙皮侧耳菌丝体,以0.6 mol/L甘露醇溶液为渗稳剂,2％溶壁酶+2％纤维素酶+1％蜗牛酶酶体系30℃酶解3h,原生质体制备率为415万个/mL；培养9d的菌丝体以0.6 mol/L硫酸镁溶液作为渗稳剂,2％溶壁酶+2％纤维素酶+2％蜗牛酶,28℃酶解4h,原生质体最佳再生率为2.86％.     

The effect of a low molecular weight heparin on coagulation parameters in healthy cats

The low molecular weight heparin (LMWH), dalteparin sodium, was administered subcutaneously (100 IU/kg) to 8 healthy cats twice daily for 13 doses. Anti-activated factor X (anti-Xa) activity was measured prior to administration (time 0), and 4, 6, 8, and 12 h after the 1st dose, 4 h after administration of the 3rd dose, and at 4, 6, 8, and 12 h after the last dose. Four cats developed measurable anti-Xa activity 4 h following a single dose, returning to baseline by 6 h. Anti-Xa activity was not detected at any time point in 4 cats. Prothrombin time (PT), activated partial thromboplastin time (APTT), and antithrombin (AT) concentrations were unaffected by LMWH administration. Dalteparin, at 100 IU/kg SC, did not achieve anti-Xa activity in 4 out of 8 cats and failed to maintain anti-Xa activity beyond 4 h in the other 4 healthy cats.     

Substances derived from 4-ethyl octanoic acid account for primer pheromone activity for the "male effect" in goats

A major constituent of the characteristic "goaty odor" 4-ethyl octanoic acid (4EOA) was previously shown to have no primer pheromone activity. This was also confirmed by our own bioassay system utilizing the recording technique of neural activity of the hypothalamic gonadotropin-releasing hormone pulse generator in goats. However, when the synthetic 4EOA solution was kept at room temperature for several months, primer pheromone activity appeared in the same solution. Headspace gas chromatography/mass spectrometry analysis revealed that there were several newly formed substances in addition to 4EOA samples with primer pheromone activity. These results suggest that 4EOA derived substance(s) but not 4EOA itself is(are) primer pheromone in goats.     

Activity of glycosidases (beta-N-acetyloglucosaminidase, alpha-mannosidase, and beta-galactosidase) in the uterine luminal fluid of cows after multiple ovulation.

This study investigated the activity of beta-N-acetyloglucosaminidase (beta-NAGASE), alpha-mannosidase, and beta-galactosidase in the uterine luminal fluid of cows after superovulation treatment, along with the possible associations between the activity of these 3 glycosidases and the superovulatory response. Embryos and a sample of fluid flushed from each uterine horn were collected on day 7 after artificial insemination (on estrus day 0) from 32 cows in which superovulation was induced with porcine follicle-stimulating hormone. Glycosidase activity was assayed colorimetrically. The cows were classified as to superovulatory response according to the number of corpora lutea per ovary (group 1, 1 to 4; group 2, > 4) and according to the total number of embryos per horn (T1, 0; T2, 1 to 2; T3, 3 to 4; T4, > 4) and the number of transferable embryos per horn (TR1, 0; TR2, 1 to 2; TR3, 3 to 4; TR4, > 4). The mean activity of beta-NAGASE was significantly lower (P < 0.05) in group 2 than in group 1, at 95.99 (standard error 20.43) versus 226.72 (46.77) IU/L. It was also significantly lower (P < 0.01) in group T4 compared with groups T1, T2, and T3, at 50.09 (8.21) versus 129.25 (34.60), 222.27 (62.62), and 290.26 (93.77) IU/L, respectively, as well as in group T1 compared with group T3. There was a positive relationship between beta-NAGASE activity and both the total number of embryos (P = 0.047) and the number of transferable embryos per horn (P = 0.013) when 1 to 4 corpora lutea developed per ipsilateral ovary. No difference in alpha-mannosidase or beta-galactosidase activity was detected among the groups.     

The effect of estrogen, progesterone and prostaglandin F2 alpha on uterine contractions in seasonally anovulatory mares

Uterine contractions were studied in two experiments utilizing ultrasonography and seasonally anovulatory mares. A one-minute ultrasound scan was done to produce longitudinal real-time images of the uterine body and an overall uterine contractile activity score (0 = no or minimal activity to 4 = maximal activity) was assigned to each scan. In experiment 1, a two-hour uterine activity trial (one score every 10 minutes) was done in mares given a single injection of prostaglandin F2 alpha (PGF2 alpha group; n = 4) and in control mares (n = 4). There was no difference between the two groups over the two-hour trial (mean activity score averaged over the two-hour trial: PGF2 alpha group, 0.2; control group, 0.1). In experiment 2, 16 mares were randomly assigned to one of four groups: 1) controls (corn oil vehicle), 2) 1 mg estradiol 17 beta on days 0 to 9 and 100 mg progesterone on days 10 to 20 (E2--greater than P4 group), 3) 100 mg progesterone on days 0 to 20 (P4 group), and 4) 100 mg progesterone on days 0 to 9 and 1 mg estradiol 17 beta + 100 mg progesterone on days 10 to 20 (P4--greater than E2 + P4 group). Uterine activity was assessed for each mare daily. The day by group interaction was significant. Scores for the E2--greater than P4 group were greater on days 4 to 11 (P less than .05) than for the other three groups. From day 14 to 21, scores did not differ among the three steroid-treated groups (except on day 15), but the scores averaged over each steroid-treated group were greater for each day (P less than .1 or .05) than for the controls (except on day 17).(ABSTRACT TRUNCATED AT 250 WORDS)     

Alterations in postmortem degradation of myofibrillar proteins in muscle of lambs fed a beta-adrenergic agonist

Dietary administration of 4 ppm of the beta-agonist L-644,969 (Merck Sharpe and Dohme Research Laboratories) to finishing lambs induced a decrease (10 to 14%, P less than .05) in extractable calpain I activity in the longissimus muscle (LD) at death (d 0). At 4 d postmortem (d 4), extractable calpain I levels in the LD of both control and treated lambs were reduced (P less than .001) from those present at d 0, but the extractable activity in the LD was reduced to a greater extent in control than in treated lambs. Calpain II activity was increased 42% (P less than .005) in LD of treated lambs; however, no significant differences were observed between d 0 and d 4 calpain II activity within treated or control LD samples (P greater than .1). Calpastatin activity was higher in the LD of treated lambs (74% on d 0, P less than .001 and 430% on d 4, P less than .001) than in the LD of control lambs. Measurable cathepsin B activity was decreased (29% on d 0, P less than .05) and measurable cathepsin H activity was increased (10% on d 0, P less than .05 and 10% on d 4, P less than .05) in the LD of treated lambs compared with controls. On d 2, 4 and 6 postmortem, degradation in myofibrils isolated from the LD was lower for treated than for control lambs. Warner-Bratzler shear values for loin chops from treated lambs were higher on both d 3 (111%) and 6 (108%) postmortem than for chops from control lambs (P less than .001). L-644,969-induced decreases in muscle proteolytic capacity may limit postmortem myofibril degradation and contribute to the reduced tenderness observed. This decreased proteolytic capacity may contribute to increased muscularity of L-644,969-treated lambs.     

高温胁迫下不同氮肥处理对高羊茅氮代谢的影响

采用盆栽试验,研究了不同氮肥（NO3--N、NH4+-N、NH4NO3-N）处理对高羊茅品种凌志Festuca arundinacea cv. Barlexas在高温38/30 ℃（昼/夜）胁迫下叶片内硝态氮含量、铵态氮含量以及参与氮同化和代谢过程的主要酶活性的影响。结果表明：随着胁迫时间的延长,经过不同氮肥处理的植株中硝态氮含量先升后降,而铵态氮含量呈上升趋势,其中NH4NO3-N处理株的硝态氮含量最高,铵态氮含量最低,NH4+-N处理株的铵态氮含量最高;不同处理株中的硝酸还原酶（NR）活性呈下降趋势,而谷氨酰胺合成酶(GS)和谷氨酸合成酶（GOGAT）活性均呈先升后降的趋势,总体上,NH4NO3-N处理株的NR、GS和GOGAT活性最高;NH4NO3-N和NO3--N处理株的谷氨酸脱氢酶（GDH）活性均呈下降趋势,而NH4+-N处理株的GDH活性呈上升趋势。在试验条件下,经NH4NO3-N处理的高羊茅受氧化胁迫程度最小,耐热性最好,有利于高羊茅越夏和延长其绿期。     

Antithrombin III activity in horses with large colon torsion

A chromogenic peptide substrate assay was used to determine serially plasma antithrombin III (AT III) activity in 4 groups of horses. Group I consisted of healthy, mature horses in which AT III activity was determined twice daily for 7 consecutive days. Groups 2 and 3 contained healthy horses in which AT III activity was monitored for 7 days after controlled, but varying, conditions of general anesthesia and surgery (median celiotomy). Group 4 was made up of patients with a presurgical diagnosis of colonic torsion. In healthy awake horses (group I), there was no difference in AT III values over time. Postoperative AT III activity in the halothane-anesthetized horses (group 2) and in the sham-operated horses (group 3) was not significantly (P = 0.05) different from base-line values at any time. A significant decrease (P = 0.05) in AT III activity was observed on postoperative days 1 through 3 in the group of horses with large colon torsion, but returned to preoperative values by day 4 after surgery in the horses that survived. In those horses that did not survive, AT III activity remained below base-line values for the duration of observation. Seemingly, plasma AT III activity in horses was not significantly affected by halothane anesthesia or surgery. Serial evaluation of AT III activity may be useful for predicting survival in horses with large colon torsion.     

Alkaline phosphatase activity in goblet cells in the small intestine of piglets experimentally infected with the coccidium Isospora suis

Alkaline phosphatase activity (EC. 3.1.3.1.) in goblet cells was investigated in the small intestine of 16 gnotobiotic piglets infected one day after delivery (DAD) by different rates of oocysts of Isospora suis coccidia. At a high infection rate of I. suis (750,000) the goblet cells were found to be highly positive to alkaline phosphatase on day 3 to day 4 after infection (DAI). In piglets infected by a low infection rate of I. suis oocysts (100,000) the activity of alkaline phosphatase activity in goblet cells was proved on days 4 to 10 after infection. In the first group of piglets, the positive goblet cells prevailed in the middle region of jejunum, with the peak on 4th DAI. It the second group of piglets a marked increase in the alkaline phosphatase activity was recorded in the goblet cells in the posterior part of jejunum on days 4 to 5 after infection and on 10th DAI. No alkaline phosphatase activity in the goblet cells was demonstrated in the control gnotobiotic piglets at the age of two to seven days.     

A porcine intrauterine 4-mda component with transforming growth factor-beta activity suppresses natural killer cell responses

A 4-MDa component, recovered from uterine luminal secretions of gilts on d 15 of pregnancy, was assessed for suppression of the lytic responses from natural killer (NK) and lymphokine-activated killer (LAK) effector cells. Each cell type originated from preparations of peripheral blood lymphocytes (PBL), and the LAK cells were generated from the incubation of PBL with interleukin-2. The PBL and LAK cells were cultured for 5 d with and without the 4-MDa component. Following culture, the cells were incubated (22 h) with NK-sensitive K-562 target cells at varying effector:target cell ratios (25:1 to 200:1). Lytic activity was assessed with the chromium-51 release assay. Additional experiments were conducted in order to determine whether suppressor activity of the 4-MDa component was time-dependent and associated with transforming growth factor-beta2 (TGF-beta2). For effector:target cell ratios combined, the 4-MDa component suppressed the lytic activity of PBL but failed to affect the LAK cells. Suppression of NK-mediated lysis occurred by d 3 of the 5-d culture period. In addition, suppressor activity of the 4-MDa component was reversed by a neutralization antibody to TGF-beta2. In conclusion, the 4-MDa component with TGF-beta2 activity suppressed the lytic responses of porcine NK cells.     

鼠防御素cryptdin 4基因在杆状病毒中的表达及活性

以防御素cryptdin4为研究对象，在昆虫一杆状病毒表达系统对cryptdin4基因的成熟肽片段进行表达。通过SDS—PAGE电泳和Western—blot检测，结果发现在相对分子质量8000处出现目的蛋白条带，表明该基因在杆状病毒中获得表达；体外抑菌试验结果显示，目的蛋白具有明显的抗金黄色葡萄球菌、伤寒沙门菌、李斯特菌等指示菌的活性。