玉米黄粉蛋白酶解过程的研究及产物分析

利用中性蛋白酶和复合风味蛋白酶对玉米黄粉蛋白进行酶解，研究了双酶法水解玉米黄粉蛋白的过程，并对水解产物进行分析。结果表明，双酶法水解玉米黄粉蛋白的最佳酶解条件为：pH＝6.7，T＝47℃，S%=0．9%，S／E复＝30，E复／E中＝8，水解时间40min，试验指标水解度(DH)％可达到32.82％，酸溶性多肽得率(YASP)％可达到87.46%，氮溶指数(NSI)％接近37.43％。经分析测定，水解产物中氨基酸含量4.84mg／g Pr，多肽含量18l.86mg／g Pr;多肽分子量主要分布于1089—1728Dal范围内。     

超临界CO2萃取对玉米醇溶蛋白提取率及水解度影响的研究

以淀粉厂副产物玉米黄粉为原料，经超临界CO2萃取脱脂脱杂后提取玉米醇溶蛋白，将玉米醇溶蛋白酶解制取玉米寡肽。采用正交试验法确定出最佳超临界CO2萃取工艺参数，即萃取压力25MPa，萃取温度45℃，萃取时间100min，CO2流量30L／h，此时脂类及异杂味物质萃取率为8．06％，萃取后玉米黄粉蛋白质含量为90．46％、脂肪含量0．1％，与未经超临界CO2萃取处理及溶剂处理的玉米黄粉相比，玉米醇溶蛋白提取率分别提高50．33％、35．69％，玉米醇溶蛋白水解率分别提高47．16％、36．75％。     

酶水解法制备大豆肽的研究

采用酶水解法，由大豆分离蛋白制备大豆肽。对五种蛋白酶水解大豆分离蛋白的特性进行了比较。筛选出水解能力最强的碱性蛋白酶，其最佳水解条件为：温度５５℃、ｐＨ１０．５、酶用量５％（Ｖ／Ｗ，相对于底物蛋白）、底物浓度５％（Ｗ／Ｖ）、反应时间６ｈ。水解度可达到３０％～４０％，产物平均肽链长度２．５～４．０。水解产物经超滤膜分离后的混合物溶解性良好，ＮＳＩ值达９８％以上。水解产物有很强烈地的苦味，用２０％的活性炭粉可以有效地吸附脱苦。脱苦的最佳条件是：温度５０℃～５５℃、ｐＨ４．０～４．５、活性炭／蛋白质＝０．１～０．２／１（Ｗ／Ｖ），慢速搅拌２ｈ。     

玉米蛋白水解条件的优化研究

本试验是以玉米黄粉为底物,采用碱性蛋白酶为水解酶进行水解。采用二次旋转回归设计,以温度、酶与底物浓度比和pH三因素进行水解,得出玉米黄粉的最佳水解条件是[S]=8%、[E]/[S]=3.3%、pH =9.85、T=45℃、水解时间(t)=4.5h。在这种水解条件下蛋白回收率>73%。     

控制酶解玉米黄粉蛋白制备富含“条件必需氨基酸”：谷氨酰胺（Gln） …

研究了醇水体系中酶法水解玉米黄粉蛋白提取Ｇｌｎ活性肽的可行性,在５０％水溶液中,２ｈ内所筛选酶的活性消失,并且２ｈ内对玉米黄粉蛋白的水解度仅有３％左右,醇浓度越高,活性降低越快,显然,除非对酶进行特殊的固定化处理,醇水体系中水解黄粉蛋白是不合适的。     

酶法水解珍珠贝肉蛋白质的工艺探讨

研究了用双酶水解技术对马氏珠母贝肉进行水解的条件。结果表明：枯草杆菌中性蛋白酶：在ｐＨ６．５、温度４０℃、酶浓度３×１０３Ｕ／ｍｌ、Ｋ＋浓度０．１２％、水解时间３．０ｈ后；再用胃蛋白酶水解：ｐＨ３．０，温度６０℃，酶浓度４．５Ｕ／ｍｌ、水解时间３．０，最适底物浓度为原料与水比例１：３进行水解，蛋白质水解率为６６．２％。水解液可用于生产海鲜调味料及功能性蛋白饮料。     

挤压膨化玉米黄粉酶解制备生物活性肽

采用挤压膨化的方法预处理玉米黄粉,可提高Alcalase酶解玉米黄粉的水解度。确定了Alcalase酶解玉米黄粉的适宜条件,即60℃,pH8.5,底物浓度5%,加酶量3%,反应60min。该条件下玉米黄粉水解度为39.54%,水解物的分子量主要分布在3819～663Da之间。Alcalase酶解玉米黄粉的水解产物具有抗氧化活性。     

玉米黄粉酶解工艺条件研究

研究木瓜蛋白酶水解玉米黄粉蛋白最佳工艺,通过单因素实验研究底物浓度、酶底比和酶解时间对制备工艺影响,并经正交实验,结果表明,木瓜蛋白酶水解玉米蛋白最佳工艺条件为:底物浓度5.0%,酶底比0.4%,pH6.5,温度50℃,进行酶解4小时水解度最高,此条件下水解度可达39.29%。     

玉米黄粉酶解提取浓缩蛋白工艺优化研究

通过单因素试验和响应曲面法,优化了采用低温α-淀粉酶酶解提取玉米黄粉浓缩蛋白的条件。所得最佳工艺为料液比1：4、酶用量36U／g玉米黄粉、温度40℃、时间90min、pH值6．2。该条件下玉米黄粉浓缩蛋白纯度达到78．96％,得率为80．13％。     

液化酶和糖化酶对生玉米的协同作用

α－淀粉酶和葡萄糖淀粉酶对玉米粉中淀粉协同水解的最佳比例为１．８ＡＧＵ／１．０ＫＮＵ，［ＡＧＵ＝葡萄糖淀粉酶单位；ＫＮＵ＝Ｋｉｌｏ Ｎｏｖｏ单位］，干磨玉米中淀粉的水解速度大于蜡质玉米淀粉和湿磨玉米中淀粉的水解速度，水解干磨玉米中淀粉比温磨玉米中淀粉效果更佳，此处指的湿磨玉米是用水或ＮａＯＨ浸泡玉米后磨碎的。 ?     

Lactation and body composition responses to fat and protein supplies during the dry period in under-conditioned dairy cows

An experiment was designed to study the effect of precalving supplementation with protein (Pr) and rumen-inert fat (F) on body composition and subsequent milk production and composition. Forty Holstein-Friesian dairy cows were allocated to 1 of 4 dietary treatments in the dry period (DP) based on a first-cut ryegrass silage, with 6 mature (in their third or greater pregnancy) and 4 young (in their second pregnancy) cows per treatment. These were low Pr, low F (silage alone); low Pr, high F (silage with 10% rumen-inert fat, mixed on a dry matter basis); high Pr, low F [silage with 5% high-protein corn gluten meal (CGM)]; and high Pr, high F (silage with 5% CGM and 10% rumen-inert fat). All the diets were individually offered ad libitum and dry matter intake (DMI) was recorded daily during the DP. After calving, all cows received ryegrass silage plus 8 kg/d of a commercial dairy concentrate. During the DP, DMI was higher for mature than for young cows. All animals recovered body condition score (0.13 units/wk, 1–5 scale), reaching a maximum score of 2.4 some days before calving. Precalving maximum muscle longissimus dorsi (LD) depth was greater for mature (47.5 mm) than for young cows (45.7 mm), and milk fat concentration was also higher for mature than for young cows (40.2 and 39.0 g/kg, respectively). Supplementation with CGM increased maximum LD depth (from 45.9 to 47.6 mm), calf birth weight (low Pr = 43.2, high Pr = 46.3 kg), and milk crude protein concentration (from 30.8 to 31.6 g/kg). Fat supplementation in the DP of the mature cows increased maximum back fat depth (from 3.6 to 4.5 mm), milk yield (low F = 26.3, high F = 28.7 kg/d), and Pr yields (low F = 837, high F = 899 g/d). Inclusion of F in the DP diets reduced casein concentration in milk at wk 3 of lactation from 26.3 to 24.5 g/kg. Milk CP yield was also increased by CGM supplementation when compared within cows receiving F-supplemented silages (low Pr, high F = 832 g/d; high Pr, high F= 877 g/d). It can be concluded that CGM supplementation in the DP increased subsequent milk Pr concentration, but milk Pr yield increased only in those animals also receiving F supplementation. Dry period diet supplementation with F increased maximum back fat depth and milk and CP yields in the mature cows, and led to more LD muscle mobilization during early lactation. Second-calving cows had a lower DMI and milk fat concentration than mature cows.     

玉米蛋白粉制备功能性短肽的研究

玉米功能短肽的制备对推动玉米蛋白资源的深加工和产业化进程有重要意义。在多次小试实验的基础上,利用酶工程技术制备玉米短肽,并对其抗氧化性进行了研究;结果表明:酶解100kg玉米蛋白粉可得玉米短肽干粉53kg,得率在50%以上,经测定,所得玉米短肽在浓度为1.67mg/mL时,其抗氧化抑制率为48.52%。     

EFFECT OF ENZYMATIC TREATMENT OF CORN GLUTEN MEAL ON LUTEIN, ZEAXANTHIN AND β-CRYTOXANTHIN EXTRACTION

Response surface methodology was used to optimize some operating parameters on the effects of alcalase treatment extraction of lutein, zeaxanthin and β -cryptoxanthin from corn gluten meal (CGM) on the yield of total xanthophylls. Three independent variables tested were enzyme concentration, substrate concentration and hydrolyzing time. The t -test and P value indicated that the quadratic of the three variables, enzyme concentration and hydrolyzing time were important linear variables affecting the total xanthophyll yield ( P <  0.005), followed by substrate concentration ( P <  0.01). The interaction effect between enzyme concentration and hydrolyzing time was also significant ( P <  0.005). Considering the efficiency, the economy of materials and the feasibility of experiment, the technique parameters were optimized by constrained complex method. The optimum conditions were obtained as follows: the enzyme concentration, the substrate concentration and the hydrolyzing time were 4,367 U/g, 11.3% and 2.4 h, respectively. The highest total xanthophyll yield was predicted of 65.69  µ g/g when CGM was hydrolyzed under the optimum condition. The suitability of the model equation for predicting the optimal response values was tested using the selected optimum conditions; the experimental yield was 65.06  ±  0.78  µ g/g, which was found to be in agreement with the predicted yield.

PRACTICAL APPLICATIONS

Corn gluten meal (CGM), a major by-product of corn wet milling, contains (on a dry basis) 200–400  µ g/g carotenoids. There are ∼840,000 tons of CGM produced in China every year. If a part of CGM produced was manufactured further to produce carotenoids as foods, materials of foods and additives of pharmaceutical products, its value and presence in the marketplace would be increased.     

玉米短肽的制备及其抗氧化活性的研究

酶解玉米蛋白对生产高营养且易于吸收、高附加值的具有生物学功能特性的生物产品具有重要意义。本文通过Alcalase AF2．4L和风味蛋白酶在适当条件下酶解玉米蛋白，酶解2h，水解度可达31％，短肽得率超过70％以上。经Sephadext^TMG-25进行分离，对其分离后的各组分进行抗氧化活性的研究，经测定，玉米短肽在浓度为1mg／ml时对邻苯三酚（PR）的自氧化抑制率达43．9％，相当于5U／ml SOD的抗氧化活性。     

剪切乳化预处理对玉米蛋白酶解反应的促进作用

研究了剪切乳化预处理对黑曲霉酸性蛋白酶水解玉米蛋白反应的影响。实验结果表明，在酸性蛋白酶与玉米蛋白粉的质量比为10％，底物浓度为80g／L，反应温度为55℃，水解24h的条件下，当玉米蛋白粉未经高速剪切乳化预处理时，蛋白质溶出率为62．79％，所得的水解产物中分子量小于1000u的寡肽占45％；而当玉米蛋白粉经7000r／min高速剪切乳化预处理1h后，蛋白质溶出率提高到88．47％，所得的水解产物中分子质量小于1000u的寡肽占78％。利用高效液相色谱法测定了玉米蛋白粉酶解产物的氨基酸组成，发现高速剪切乳化预处理能增加水解产物中支链氨基酸的含量，有利于生产高F值寡肽。     

In vitro antioxidant activity of protein hydrolysates prepared from corn gluten meal

BACKGROUND: Corn gluten meal (CGM), a major by‐product of corn wet milling, is mainly used as forage in China. Because of its particular amino acid composition, in which there are large amounts of hydrophobic amino acids such as leucine, alanine and phenylalanine, CGM protein was thought to be a good resource to obtain antioxidant peptides. CGM protein was hydrolysed with a biochemical grade alcalase and the derived hydrolysates were assessed for their antioxidant properties in different in vitro assay systems, including inhibiting activity on lipid peroxidation, by reducing power, hydroxyl radical scavenging activity, and DPPH radical scavenging activity. The effects of concentration and molecular weight (MW) of hydrolysates on antioxidant activity were investigated. RESULTS: The results showed that CGM hydrolysates were effective antioxidants, and there was a dose‐dependent relationship between hydrolysate concentration and antioxidant activity; the highest antioxidant activity was found in peptides 500–1500 Da, and the antioxidant activity of peptides below 500 Da or peptides above 1500 Da were all lower than that of total hydrolysates. CONCLUSIONS: The finding showed that the antioxidant activity of CGM hydrolysates was related to molecular weight and hydrolysate concentration, and the active antioxidant fraction should be in the peptides fraction of 500–1500 Da. CGM protein hydrolysates can be a source of natural antioxidant and used as a food additive. Copyright © 2008 Society of Chemical Industry     

Effects of feeding dairy cows protein supplements of varying ruminal degradability

Twenty-five (10 ruminally cannulated) Holstein cows averaging 82 +/- 34 d in milk were assigned to 5 x 5 Latin squares (21-d periods) and fed diets supplemented with one of four different proteins to assess effects on production, ruminal metabolism, omasal flow of N fractions, and degradation rates of protein supplements. Total mixed diets contained (dry matter basis) 44% corn silage, 22% alfalfa silage, 2% urea, and 31% concentrate. Five concentrate mixes were fed: 31% high-moisture shelled corn (HMSC; basal); 9% solvent soybean meal (SSBM), 22% HMSC; 10% expeller soybean meal (ESBM), 21% HMSC; 5.5% blood meal (BM), 25.5% HMSC; and 7% corn gluten meal (CGM), 24% HMSC. Diets averaged, respectively, 15.8, 19.1, 19.7, 20.3, and 19.3% crude protein. Feeding the basal diet reduced intake and yield of milk, fat-corrected milk (FCM), and all milk components compared to the protein-supplemented diets. Milk yield was higher for cows fed ESBM and CGM, fat yield was higher for cows fed SSBM and CGM, but FCM and protein yields were not different among cows fed supplemental protein. Based on omasal sampling, mean in vivo estimates of ruminal degradation rate for the crude protein in SSBM, ESBM, BM, and CGM was, respectively, 0.417, 0.179, 0.098, and 0.051/h (computed using passage rates observed for the small particle phase; mean = 0.14/h), and 0.179, 0.077, 0.042, and 0.026/h (computed using a passage rate of 0.06/h). The in vivo degradation rate computed for SSBM at a passage rate = 0.06/h was similar to that estimated using the inhibitor in vitro method. However, in vivo degradation rates computed at passage rate = 0.06/h for ESBM, BM, and CGM were about two, four, and three times more rapid than those estimated by inhibitor in vitro. Experimental proteins fed in this trial will be used as standards for developing in vitro methods for predicting rates of ruminal protein degradation.     

Pilot-scale production of low molecular weight peptides from corn wet milling byproducts and the antihypertensive effects in vivo and in vitro

A pilot-scale production was developed to enhance the value of proteins from corn gluten meal (CGM). Corn protein isolate (CPI) with high protein content (90.68%) was obtained through heat treatment of CGM (150 kg) in aqueous alkaline solution. Two-step enzymatic hydrolysis and multistage separation were applied to enrich corn oligopeptides (COP) with low molecular weights, 96.77% of which were less than 1000 Da. The greatest antihypertensive effect of COP treatment in spontaneously hypertensive rats (SHR) was observed at a dose of 0.45 g/kg. One major ACE-inhibitory peptide, Ala-Tyr, was identified and quantified (9.16 ± 0.08 mg/g) from COP. The ACE inhibitory activity of Ala-Tyr (IC₅₀ = 0.037 mg/ml) was over 27 times higher than that of COP (IC₅₀ = 1.020 mg/ml). The results indicate that COP may be a source of natural antihypertensive compounds that could be used for drugs or functional food ingredients.     

Dietary preference in dairy calves for feed ingredients high in energy and protein

In 3 experiments, we assessed preference of recently weaned dairy calves for (1) 8 high-energy feed types [barley meal, corn meal, corn gluten feed (CGF), oat meal, rice meal, sorghum meal, wheat meal, and wheat middlings meal]; (2) 6 high-protein feed types [corn gluten meal (CGM), wheat distillers dried grains, rapeseed meal, soybean meal (SBM), sunflower meal, and pea meal]; and (3) 4 mixtures (50:50) of the highest- and lowest-ranked high-energy and high-protein feeds, to assess whether calves maintain preference for feed ingredients that are included in a mixture. In all experiments, pairwise preference tests were conducted between all feed types (28 different pairwise preference tests in experiment 1, 15 tests in experiment 2, and 6 tests in experiment 3). Each pairwise preference test was conducted by offering ad libitum access to both feed types for 6 h. All tests were repeated with 20 Holstein calves. Before this study, calves were offered milk replacer at a rate of 4 L/d and a pelleted starter feed ad libitum. After weaning at 62 d of age, each calf was involved in a pairwise preference test at 3 and 5 d postweaning. A preference ratio was calculated for each calf in each test as (intake of feed type A)/(intake of feed type A + intake of feed type B). Preference for feed types was ranked across tests in each experiment using pairwise comparison charts. In experiment 1, the highest-ranked high-energy feed type was wheat meal and the lowest ranked were rice meal and CGF. In experiment 2, the highest-ranked high-protein feed type was SBM and the lowest ranked was CGM. According to the preference rankings from experiments 1 and 2, experiment 3 evaluated (50:50) mixtures of SBM + wheat meal, SBM + CGF, CGM + wheat meal, and CGM + CGF. The mixture of SBM + wheat meal was highest ranked, CGM + CGF was lowest ranked, and the mixtures containing one high-ranked and one low-ranked feed ingredient (SBM + CGF and CGM + wheat meal) were ranked equally. The results of this study indicate that young calves exhibit clear preferences for certain high-energy and high-protein feeds that may be considered highly palatable. Further, preference ranking of feed types provided as 50:50 mixtures was consistent with ranking of individual feed types, suggesting that palatability of mixed starter rations can be improved by inclusion of a preferred feed type.     

超声预处理对玉米蛋白可酶解性的影响

旨在采用超声预处理改善玉米蛋白的酶解性,研究了超声频率模式(单频、双频、定频和扫频)及频率参数对玉米蛋白酶解水解度及溶解率的影响,采用高效尺寸排阻色谱法对酶解液的分子质量分布进行了表征。研究发现,(68±2)kHz/(28±2)kHz双频扫频为最佳超声频率模式,优化试验获得超声预处理的最佳工作参数为:超声温度30℃、料液比1:20(g/mL)、超声处理时间40 min、扫频周期500 ms、超声功率密度80 W/L、脉冲超声的工作时间10 s和间歇时间3 s。在最佳超声频率模式及工作参数预处理的条件下,水解度和蛋白溶解率分别为23.6%和75.2%,较对照组(未经超声处理)分别提高了39.4%和54.7%,且酶解液具有更窄的分子质量分布范围。这表明超声预处理能促进玉米蛋白向多肽转化,尤其有利于分子质量为200~1 000 u的玉米蛋白肽的形成。