微波辅助水解比色法快速检测羊奶粉中水解动物蛋白

目的：寻求一种快速、高效测定羊奶粉中是否添加水解动物蛋白（HAP）的方法。方法：以L-羟脯氨酸为水解动物蛋白特征物,样品通过微波辅助水解处理,测定样品羟脯氨酸的含量变化,并与传统湿法水解对比。结果：微波辅助水解条件：硫酸浓度4mol/L,功率720W,时间45min时,HAP及掺假合成品水解后测得的羟脯氨酸含量与传统湿法水解所测得的羟脯氨酸含量基本相同,RSD为-4.5%～1.6%。本方法可以快速判定羊奶粉中是否添加水解动物蛋白,具有较高准确度和重现性,羟脯氨酸含量与吸光度具有良好的线性关系,决定系数R2=0.9968,加标回收率86.24%～91.30%,可以满足比色法测定结果要求。结论：微波辅助水解可以将样品前处理时间从16h缩短到45min,此方法快速简便,重复性好,回收率高。     

新疆双峰驼乳酪蛋白血管紧张素转化酶抑制肽的酶法制备和抑制特性

对新疆双峰驼乳酪蛋白分别进行胃蛋白酶和胰蛋白酶的单酶和双酶联合水解,用反相高效液相色谱外标法,对产生的马尿酸含量进行检测,测定水解产物血管紧张素转化酶(angiotensin converting enzyme,ACE)抑制活性.通过米式方程对比水解产物与卡托普利之间的竞争关系,并用50、10、3?kDa的超滤膜对水解...     

高效液相色谱-蒸发光散射检测法与酶法检测UHT灭菌乳中乳果糖含量的对比研究

采用液相色谱法分离、蒸发光散射检测器检测,外标法定量测定超高温瞬时灭菌乳中乳果糖的含量。采用盐酸沉淀蛋白,氨基色谱柱分离,流动相为75％乙腈溶液。该方法能使乳果糖得到很好分离,精密度为0.77％(n=6),平均回收率为98.7％～102.6％。高效液相色谱-蒸发光散射检测法较酶法操作简便、省时、重复性好,可以根据实验需求选择使用。     

微波水解—氨基酸自动分析仪法测定饲料中色氨酸的含量

[目的] 建立微波水解—氨基酸自动分析仪快速测定饲料中色氨酸含量的方法。[方法] 以奶牛全混合日粮饲料（TMR）为研究对象,通过单因素试验和正交试验筛选微波碱水解最佳条件,联合氨基酸自动分析仪LCAK06/Na（4.6 mm×150 mm）色谱柱分离及快速测定饲料中色氨酸含量。[结果] 水解温度是影响饲料色氨酸含量的最主要因素。微波碱水解测定饲料中色氨酸含量的最佳条件是：水解温度170 ℃、水解时间30 min、碱浓度5 mol/L,通过氨基酸自动分析仪测定TMR饲料色氨酸含量为0.14%。色氨酸在0.025~0.600 μmol/mL峰面积与浓度线性关系良好,相关系数（r）为0.999 7。定量限（LOQ）为0.05%,色氨酸回收率93%~104%,精密度RSD为3.58%。[结论] 该方法高效、准确,检测结果与国家标准《饲料中色氨酸的测定》（GB/T 15400—2018）中的常规碱水解法对比无显著差异,对于饲料中色氨酸含量测定具有参考价值。     

不同家蝇幼虫制品对黄羽肉仔鸡营养物质可利用率、肠道菌群和血清生化指标的影响

本研究旨在探讨不同家蝇幼虫制品对黄羽肉仔鸡营养物质可利用率、肠道菌群和血清生化指标的影响。选取 １日龄的黄羽肉仔鸡 ２５２只,随机分成 ７组,每组 ３个重复,每个重复 １２只鸡。对照组饲喂基础饲粮,其余 ６组饲喂分别用 ０．２％、０．５％的家蝇幼虫肽,０．２％、０．５％的家蝇幼虫酶解物,０．２％、０．５％的脱脂家蝇幼虫粉等量替代基础饲粮中鱼粉的试验饲粮。结果表明：１）添加家蝇幼虫肽有提高干物质、粗蛋白质和能量的可利用率的趋势（Ｐ＞０．０５）,且随着添加剂量的增加而提高;添加 ０．５％的家蝇幼虫肽可显著提高粗脂肪、钙、磷和粗灰分的可利用率（Ｐ＜０．０５）。２）添加家蝇幼虫肽可显著提高试验第２周和第４周肠道乳酸杆菌和双歧杆菌的数量（Ｐ＜０．０５）,显著降低大肠杆菌和沙门氏菌的数量（Ｐ＜０．０５）。３）添加家蝇幼虫肽可显著降低试验第 ２周血清谷丙转氨酶活性（Ｐ＜０．０５）和显著提高血清总蛋白含量（Ｐ＜０．０５）,且以０．５％的家蝇幼虫肽组效果最优。在饲粮中添加不同家蝇幼虫制品均能显著降低黄羽肉仔鸡血清尿素氮含量（Ｐ＜０．０５）。由此得出,家蝇幼虫肽具有改善黄羽肉仔鸡肠道菌群、提高营养物质可利用率的作用,且以 ０．５％的添加量效果较好。     

鸭血球短肽的优化制备及其特性研究

本试验旨在筛选水解效率高且脱色效果好的商业蛋白酶,建立血球短肽的优化工艺,比较其在酶解前后营养特性的变化,研究其功能特性与体外抗氧化能力,以研发功能性血球短肽产品,为家禽血液资源高值化的转化利用与深度挖掘提供理论依据与技术借鉴。比较酶种类、酶浓度、温度、p H、水解时间等因素对蛋白酶水解度(DH)、脱色程度、水解物产量的影响,采用正交试验设计优化血球短肽的最佳工艺,对血球短肽进行营养价值、功能特性及抗氧化性能评价。确定酸性蛋白酶为最佳水解酶,其水解鸭血球蛋白制备短肽的最优工艺参数为:酶用量6 000 U/g,温度50℃,p H 3.5,水解时间7.0 h。在此条件下,水解度为(25.10±0.65)%,水解物产量为(60.09±1.77)%。通过高效液相色谱分析水解产物分子质量分布。结果表明,酶解对血球蛋白有明显的降解作用,酶解产物主要以3 ku以下的短肽为主,其中1 ku以下占大部分(62.82%)。血球短肽粉呈乳白色,氨基酸种类齐全,必需氨基酸含量丰富(53.31%),鸭血球蛋白酶解后的溶解性大大提高(60%),且具有良好的乳化稳定性。血球短肽清除自由基能力较强,随血球蛋白浓度的提升,清除1,1-二苯基-2-苦基肼自由基(DPPH·)与超氧阴离子能力随之愈强,还原力也逐渐增加。由此可见,酸性蛋白酶可有效水解鸭血球蛋白获得氨基酸含量丰富、溶解性好且具有抗氧化活性的乳白色血球短肽,可以作为功能性原料应用于食品与饲料中。     

母乳乳清蛋白抗氧化活性肽的研究

母乳是婴幼儿的最佳食物来源,而乳清蛋白是营养和活性物质的基础,其中生物活性肽对人体健康具有重要促进作用.为研究母乳乳清蛋白抗氧化活性肽,采用中性蛋白酶、碱性蛋白酶、木瓜蛋白酶和胰蛋白酶4种酶制备抗氧化活性多肽,通过单因素试验和响应面分析对乳清蛋白酶解工艺进行优化.结果表明:中性蛋白酶最适于母乳乳清蛋白抗氧化肽的制备,此时的最佳工艺参数为pH 7.21、反应温度50.03℃、酶与底物比(E/S)4 486.68 U/g、酶解时间5h;影响1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率的因素大小为:酶与底物比＞温度＞pH值.利用大孔树脂、葡聚糖凝胶过滤色谱G-25、G-15分析得出组分峰Ⅰ抗氧化活性最强,其DPPH自由基清除率达到了60.31％.     

犬类诱食剂中酶解反应与美拉德反应最佳条件优化研究

试验旨在开发一款适用于干犬粮的诱食剂,研究犬类诱食剂中酶解反应与美拉德反应的最佳条件.试验选用鸡胸肉、鸡心、鸡肝为主要原料,采用木瓜蛋白酶进行水解,在酶解液中添加木糖和L-半胱氨酸,利用美拉德反应产物制备适合犬类的诱食剂.采用单因素和正交试验,以水解度为考核指标,研究水解反应过程;通过感官评价及动物试验探索最佳美拉德反...     

菜籽饼的发酵脱毒及酶酸法水解制备复合氨基酸工艺研究

以菜籽饼为原料,选择黑曲霉、米曲霉、乳酸菌进行混合固态发酵,然后利用多菌种混合发酵中产生的蛋白酶进行菌酶水解试验,经55℃菌酶水解7 h,氨基酸含量达1.15 g/100 ml;然后,加适量盐酸,经95℃低酸水解8 h,氨基酸含量可达2.17 g/100 ml。本项目研究的生产复合氨基酸液的工艺集脱毒、菌酶、低酸水解为一体,能耗低、产率高,且氨基酸种类丰富齐全,为菜籽饼的综合利用开辟了一条新的途径。     

Hydrolysis of phytic acid by intrinsic plant and supplemented microbial phytase (Aspergillus niger) in the stomach and small intestine of minipigs fitted with re-entrant cannulas. 3. Hydrolysis of phytic acid (IP6) and occurrence of hydrolysis products (IP5, IP4, IP3 and IP2)

Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (Aspergillus niger) were investigated with six minipigs (40-50 kg initial body weight) fitted with re-entrant cannulas in the duodenum, 30 cm posterior to the pylorus (animals 1, 4, 5 and 6) and ileocecal re-entrant cannulas, 5 cm prior the ileocecal junction (animals 1, 2 and 3), respectively. Dietary treatments were as follows: (1) diet 1, a corn-based diet [43 U phytase/kg dry matter (DM)]; (2) diet 2, diet 1 supplemented with microbial phytase (818 U/kg DM); and (3) diet 3, a wheat-based diet (1192 U/kg DM). At 07 30 h and 19 30 h, each animal was fed 350 g diet mixed with 1050 ml de-ionized water. Digesta were collected continuously and completely during a 12-h period after feeding. Mean hydrolysis rates of IP6 in the stomach as measured at the proximal duodenum of animals 1, 4, 5 and 6 were 9.0, 77.2 and 66.2% for diet 1, 2 and 3, respectively. Microbial phytase was much more effective in phytate hydrolysis than wheat phytase. Mean IP6 hydrolysis rates of the respective diets in the stomach and small intestine as measured at the distal ileum of animals 1, 2 and 3 were 19.0, 62.6 and 64.6% and were lower than treatment means of the stomach only. Differences existed between experimental animals with respect to their ability to hydrolyse IP6 in the stomach independent of the presence and source of dietary phytase. Considerable amounts of hydrolysis products occurred in both the duodenal and ileal digesta when diets 2 and 3 were fed; however, only traces were determined after ingestion of diet 1. Independent of dietary treatment, four IP5 isomers were detected, but in different amounts.     

L-Cys/EGDE对水解老化和光老化真丝织物的加固效果

采用L-半胱氨酸(L-Cys)和乙二醇二缩水甘油基乙醚(EGDE)作为加固材料,分别对水解老化和光老化真丝织物进行加固处理,为脆弱丝绸文物的加固保护提供简便而有效的技术方法。水解老化样品在加固后的断裂强力由1.9 N±0.3 N提高到18.0 N±0.4 N,断裂伸长从3.0 mm±0.5 mm提高到14.0 mm±0.5 mm;而光老化样品加固后断裂强力由1.8 N±0.3N提高到4.1 N±0.5 N,断裂伸长保持1.0 mm±0.3 mm不变。通过热重分析、13C固体核磁共振表征发现,水解老化样品加固后的热稳定性有所提高,分子结构有所变化;而光老化样品加固后的热稳定性及分子结构都没有发生明显变化。氨基酸分析证明真丝织物在光老化处理后,可以与EGDE反应的酪氨酸(Tyr)、组氨酸(His)的含量大大降低,造成反应位点的大量减少,因此与水解老化样品的加固效果悬殊。将真丝织物水解老化样品、光老化样品的丝蛋白氨基酸组成及含量与文物样品进行比较发现,水解老化样品的丝蛋白氨基酸组成及含量与文物样品更为接近,因此,可利用该加固方法对脆弱丝绸文物进行加固保护。     

燕麦秸秆纤维素测定方法比较及响应面优化分析

为比较测定燕麦秸秆纤维素含量的不同方法,试验运用浓硫酸水解、蒽酮比色、聚酯纤维滤网袋、纤维素分析仪测定法,分别对3个生态型燕麦秸秆进行纤维素含量测定,并对比找出较优测定法。方差分析结果表明:较好的测定方法为浓硫酸水解法,但缺点是测定的纤维素含量偏低,需进一步改进。试验针对浓硫酸水解法,选取其药品筛目数、加热时间、清洗次数、消煮时间作为改进条件,对XO-1-03生态型燕麦秸秆进行纤维素含量测定。利用Design Expert 8.0.5b(响应面法)分析得出最佳条件为:孔径药品筛30目、洗涤次数2次、水浴时间30 min、消煮时间12.5 min。在此条件下利用浓硫酸水解法测得纤维素含量较高、稳定性较好。     

A step-wise in vitro method to estimate kinetics of hydrolysis of feeds

Nutritional feed values are currently based on aggregate criteria such as the ileal or faecal digestibility. Digestibility is the result of two competing processes: digestion and passage. In order to develop mechanistic models of digestion to be used for feed evaluation, both processes have to be quantified. The aim of this study was to determine the hydrolysis kinetics of main constituents (organic matter, N and starch) in wheat, barley, wheat bran and soybean meal, using a three-step, enzymatic in vitro method mimicking digestion in the stomach, small and large intestine of pigs. Kinetics of hydrolysis (i.e. solubilisation of feed constituents) was modelled using an exponential segmented model estimating the extent and rate of digestion in each digestive segment. Digestion of organic matter of soybean meal occurred mainly in the stomach (67% vs. < 30% for other feeds) resulting from an efficient protein hydrolysis at this site. Organic matter of cereals was mainly digested in the small intestine (55%), as a result of starch digestion. Fractional digestion rates of organic matter were higher in the stomach than in the small intestine (20–35 vs. 2–15%/min). The potential digestibility of organic matter was nearly 87% for wheat and soybean meal, 78% for barley and 61% for wheat bran, which correspond to typical in vivo digestibility values. The results of the present study allow characterising different feeds with the objective to model the digestion process in pigs.     

糠氨酸标准溶液的储存稳定性以及盐酸浓度对牛奶中糠氨酸检测结果的影响

本试验旨在研究糠氨酸标准溶液的储存稳定性以及牛奶前处理过程中盐酸浓度对糠氨酸检测结果的影响.试验1:分别用3.0、1.0和0.1 mol/L盐酸配制不同浓度糠氨酸标准溶液(1和5 mg/L),并分别放置于-20、4℃和室温(25℃)下,于第1、3、5、8、10、12、14、15、17和21天采用超高效液相色谱法测定糠氨...     

Serum lipase determination in the dog: a comparison of a titrimetric method with an automated kinetic method

An enzymatic, kinetic method for determining serum lipase activity was evaluated and compared to a standard manual method for use in dogs. The kinetic method was a commercial kit adapted for use on a tandem access clinical chemistry analyzer and utilized a series of coupled enzymatic reactions based on the hydrolysis of 1,2-diglyceride by lipase. The manual method was the Cherry-Crandall technique based on the titration of base against the acid formed by hydrolysis of an olive oil substrate by lipase. The correlation between the two methods was very good (r = 0.94). The reference range for 56 clinically healthy dogs assayed by the kinetic method was 90 to 527 U/L. Diseases associated with a greater than twofold elevation in serum lipase activity as determined by the kinetic method included pancreatitis, gastritis with liver disease, and oliguric renal failure with metabolic acidosis. In some cases, pancreatitis was seen with other clinical problems, such as gastroenteritis, diabetic ketoacidosis, duodenal mass, disseminated intravascular coagulation, and septic peritonitis. Diseases associated with serum lipase activity within the reference range or elevated less than twofold included gastritis, gastric ulcer, cholestasis, phenobarbital-induced hepatopathy, colitis, copper hepatopathy, abdominal hematoma, apocrine gland adenocarcinoma, and thrombocytopenia with pneumonia.     

木聚糖酶产生菌的筛选鉴定及木聚糖酶基因(Xyn B)的克隆

采用富集培养、透明圈平板筛选的方法,从长期堆放青贮饲料的土壤中分离到7株木聚糖酶产生菌。选取透明圈最大的木聚糖酶产生菌X7作为基础菌,对其进行16S rRNA部分序列PCR扩增测定,经BLAST同源性分析确定X7为枯草芽孢杆菌Bacillus subtilis strain GD3b(Genebank编码:HM055602.1)。然后设计特异引物,以X7基因组为模板,对其进行Xyn B基因序列PCR扩增,扩增产物经BLAST同源性分析表明已成功克隆了X7菌株的Xyn B基因,为日后构建木聚糖酶基因酵母表达载体,培育能够分泌表达具有高活性的木聚糖酶酵母工程菌奠定了基础。     

Colorimetric assay for the quantification of serum iodixanol concentration and its application on estimation of glomerular filtration rate in cattle

A colorimetric assay used to quantify the non-ionic contrast medium iodixanol in sera was validated and compared with high-performance liquid chromatography. The application of this assay to estimate glomerular filtration rates (GFR) in cattle was examined. Serum iodixanol was de-iodinated by alkaline hydrolysis and the amount of released iodine was subsequently determined using a ceric arsenite method. There was a close correlation between the two methods using identical specimens. In clinically healthy cattle with different body weights, the reference value (166.3-178.8 mL/min/m(2)) based on body surface area was fairly stable as compared with that (2.13-3.63 mL/min/kg) based on body weight. Based on GFR data in healthy and renal-impaired cattle, when the GFR decreased to more than 60% of the reference value, serum urea nitrogen and creatinine concentrations increased. The colorimetric assay is a simple method for the estimation of GFR in cattle and requires no expensive equipment.     

A metallo phosphatase activity present on the surface of Trypanosoma brucei procyclic forms

In this work, we describe how living cells of Trypanosoma brucei procyclic forms were able to hydrolyze extracellular p-nitrophenylphosphate (pNPP). These intact parasites, which had their viability determined by motility and the Trypan blue method, presented a low level of pNPP hydrolysis in the absence of any divalent metal (0.72+/-0.07 nmol pNP/mg min). Interestingly, in the presence of 5mM MgCl(2), ectophosphatase activity of 1.91+/-0.21 nmol pNP/mg min was observed. The ectophosphatase activity was also stimulated by MnCl(2), CoCl(2) and CuCl(2) but not by CaCl(2) and CdCl(2) and was inhibited by ZnCl(2). The addition of Mg(2+), Mn(2+), Co(2+) and Cu(2+) to extracellular medium increased the ectophosphatase activity in a dose-dependent manner. At 5mM pNPP, half-maximal stimulation of pNPP hydrolysis was obtained with 0.39+/-0.05 mM MgCl(2), 0.33+/-0.03 mM MnCl(2), 1.63+/-0.12 mM CoCl(2) and 2.04+/-0.33 mM CuCl(2). In the absence of any divalent metal (basal activity) the apparent K(m) for pNPP was 0.66+/-0.09 mM, while at saturating MgCl(2) concentrations the corresponding apparent K(m) for pNPP for Mg(2+)-stimulated phosphatase activity (difference between total minus basal phosphatase activity) was 0.27+/-0.03 mM. The Mg(2+)-stimulated pNPP hydrolysis was strongly inhibited by ZnCl(2) and vanadate, while the metal-independent basal phosphatase activity was less inhibited by these phosphotyrosyl phosphatase inhibitors.     

酶解日本对虾副产物主要成分及其可溶性肽组成分析

为确定酶解日本对虾(Penaeus japonicus)加工副产物的主要成分及其可溶性肽相对分子质量组成,采用多酶分步水解技术制备日本对虾加工副产物的酶解产物(EHP)。用高效液相色谱法、紫外可见分光光度法、索氏法、化学分析法等方法,分析EHP主要营养组成及其肽的相对分子质量。结果表明,EHP含蛋白质67%、粗脂肪1.0%、类胡萝卜素0.12%、钙6.7%、磷0.4%。EHP中的最长肽链相对分子质量为13 800,仅占0.32%;相对分子质量在5 400~1 550的肽占35.72%、相对分子质量小于1 000的小肽占52%。每100 g EHP干基中的总氨基酸含量50.185 g、人体必需氨基酸17 g,鱼虾10种必需氨基酸22 g,鲜味氨基酸24.7 g。     

酶解羽毛粉的体外蛋白质消化率及其在生长猪日粮中的应用效果

试验旨在测定酶解羽毛粉的体外蛋白质消化率,同时研究酶解羽毛粉替代日粮中鱼粉对生长猪生产性能和蛋白质消化率的影响。体外试验采用胃蛋白酶-胰酶水解法对水解羽毛粉和酶解羽毛粉进行消化测定其蛋白质消化率。饲养试验选用135头体重相近的杜×长×大生长猪,随机分成3组,分别为对照组(3%秘鲁鱼粉组),处理Ⅰ和Ⅱ(酶解羽毛粉50%和100%替代秘鲁鱼粉)。结果表明:酶解羽毛粉体外蛋白质消化率比水解羽毛粉提高了15.38%(P<0.05)。处理组平均日增重和平均日采食量与对照组均无显著差异(P>0.05);饲料增重比及蛋白质消化率也与对照组差异不显著(P>0.05)。结合生产性能试验与经济效益分析,酶解羽毛粉100%等蛋白替代秘鲁鱼粉组为最佳,在生产实践中具备实用性和可行性。