【作者】 玛丽娜；

【导师】 敖日格乐；

【作者基本信息】 内蒙古农业大学 ， 动物营养与饲料科学， 2014， 博士

【摘要】 乳清是用牛乳生产干酪、干酪素过程中所产生的副产品,具有乳糖、乳清蛋白、矿物质、维生素等多种营养成分,且具有多种生理功能特性。但是,由于乳清中干物质相对含量低,开发利用技术复杂,设备昂贵,大量的乳清往往被干酪生产企业当作废水排泄掉,不仅造成资源的浪费,也污染了环境。因此,为了科学的开发和利用乳清资源,本论文研究了乳清的深度发酵制品—功能性乳清醋。功能性乳清醋不仅符合当前人们日益增强的营养保健需求,而且还可以将干酪工业副产物变废为宝,为干酪工业乳清的去向问题提供了新思路。目前有关乳清醋的研制问题国内未见有相关报道,国外这方面的研究报道也较少,论文研究了乳清醋的发酵工艺条件,并且对其风味物质及部分营养成分进行了分析研究。通过动物试验研究,揭示了本研究制备的乳清醋具有显著增强免疫力的功能。具体研究方法和结果如下：1.乳清醋的发酵试验采用Plalckett-Burman试验设计,确定影响酒精发酵及醋酸发酵过程的重要因素,采用最速上升试验方法确定影响酒精发酵及醋酸发酵过程的最优因素水平,最后用B6-Behnken试验设计确定了两个发酵过程的最优发酵条件。试验结果如下,酒精发酵条件为：A马克斯克鲁维酵母接种量为4×106cfu/ml;B酿酒酵母接种量为4.5×106cfi/ml;C发酵时间为151h；D鼠李糖乳杆菌接种量为2×106cfu/ml;E德氏乳杆菌接种量为2×106cfu/ml;F温度为40℃；G糖度为14%；H初始pH6.1；I姜粉用量为0.095(g/100ml)；醋酸发酵条件为：A温度为30℃；B初始为pH5.7:C酒精度为7.0；D发酵时间617；E醋杆菌接种量为6.5×106cfu/ml;F巴氏醋杆菌接种量为6.5×106cfu/ml;2.风味物质及营养成分分析试验采用顶空固相微萃取-气质联用技术对乳清醋的挥发性风味成分进行了分析,试验共测得36种化合物(匹配度80%以上),7大类化合物,其中酯类物质5种,酸类5种,酮类4种,醇类10种,醛类6种,酚类2种,烷烃类4种。通过氨基酸分析仪分析游离氨基酸含量发现25种氨基酸,其中包含8种人体必需氨基酸和4种具有增强免疫功能作用的氨基酸,有赖氨酸、苏氨酸、精氨酸、丝氨酸等。通过对乳清醋中矿物质及维生素的试验测定,在16种成分中,除了乳清醋当中氯和钠的含量远低于白米醋和苹果醋,Fe含量略低于白米醋和苹果醋,其它13种成分(钾、钙、镁、锌、锰,碘、磷、硒、VB1，VB2、Vc、VA、VE)的含量均高于白米醋和苹果醋。3.通过动物试验研究了乳清醋对小鼠免疫功能的促进作用,具体内容包括以下几方面：(1)脏器／体重比值测定试验：与对照组相比,高、中两个剂量组的乳清醋能显著增强小鼠的免疫器官重量、胸腺指数及脾脏指数(P<0.05)。(2)二硝基氟苯诱导小鼠迟发型变态反应试验：与对照组相比,高、中、低三个剂量组的耳肿胀度均有显著性差异(P<0.05),说明乳清醋能显著提高小鼠迟发型超敏反应。(3) ConA诱导的小鼠淋巴细胞转化试验：与对照组相比,高、中、低三个剂量组的乳清醋对促进淋巴细胞增殖均有极显著地影响(P<0.01)。.(4)抗体生成细胞检测：与对照组相比,乳清醋各剂量组对小鼠抗体细胞生成没有显著地影响。(5)血清溶血素测定试验：与对照组相比,乳清醋各剂量组对小鼠的血清溶血素没有明显的影响,抗体生成水平也无显著性差异。(6)小鼠腹腔巨噬细胞吞噬鸡红细胞试验：与对照组相比,高、中两个剂量组的乳清醋对增强小鼠巨噬细胞的攻击和吞噬能力有极显著地影响(P<0.01)。(7)小鼠中的碳廓清试验：与对照组相比,高、中、低三个剂量组的乳清醋均可显著增加小鼠的吞噬指数。(P<0.05)(8)NK细胞活性测定：与对照组相比,高、中、低三个剂量组的乳清醋对增强NK细胞活性有极显著地影响。(P<0.01)本研究论文得出的结论是,以乳清为原料,采用液态发酵方法,经过酒精发酵和醋酸发酵过程可成功制得乳清醋。乳清醋含有的营养物质丰富于普通白米醋和苹果醋,且风味物质浓厚。经过动物试验证明乳清醋具有增强小鼠免疫功能的作用。更多还原

【Abstract】 Milk whey is the byproduct of producing cheese and casein. It contains nutrients, including lactose, whey protein, minerals and vitamins, and has important physiological features. However, due to the low concentration of dry matter in the milk whey, the complexity of its development and utilization, and the high cost of the equipment used in the concentration process, many cheese producers often discharge milk whey as wastewater. This is not only a waste of resources, but is also an environmental pollutant. Therefore, to scientifically develop and utilize milk whey resources, this dissertation discusses the milk whey product that results from deep fermentation, that is, milk whey vinegar.This product will not only meet consumers’increasing demand for healthy foods, but also provide a new solution for the cheese production industry, by manufacturing a valuable product from a byproduct of the cheese-making process. Currently, there is very little research conducted on the development of milk whey worldwide. Therefore, the study discussed in this dissertation contributes significantly to the growth of the milk whey industry.This dissertation discusses milk whey fermentation techniques and analyzes the flavor substances and some of the nutritional components of whey vinegar. Additionally, laboratory tests show that whey vinegar has a significant effect on building up the immune system of rats. The research study methodology and results are as follows:1. The liquid-state fermentation method was adopted to produce whey vinegar. The Plackett-Burman Test was employed to determine the factors that play important roles in the fermentation process, both at the alcohol and acetic acid stages. The Fast-Rising Test was used to determine the most optimal factors at the alcoholic and acetic acid stages of the fermentation process. The Box-Behnken Test was employed to determine the two optimal conditions for the fermentation process. Test results show that the optimal conditions for alcohol fermentation are:(A) a dosage of kluyveromyces marxianus4x106cfu/ml,(B) a dosage of saccharomyces cerevisiae4.5×106cfu/ml,(C) fermentation time151h,(D) a dosage of lactobacillus rhamnosus2×106cfu/ml,(E) a dosage of lactobacillus delbrueckii subsp. bulgaricus2×106cfu/ml,(F) temperature40℃,(G) sugar concentration40%,(H) initial pH6.1, and(I) ginger power0.095(g/100ml). The optimal conditions for the acetic fermentation are:(A) temperature30℃,(B) initial pH5.7,(C) ethanol concentration7.0,(D) fermentation time617h,(E) a dosage of acetobacter sp.6.5×106cfu/ml, and(F) a dosage of acetobacter pasteurianus subsp.6.5×106cfu/ml.2. The Solid-Phase Micro-Extraction Fibers and the Gas Chromatography-Mass Spectrometry (GC-MS) tests were performed to extract the volatile compounds of whey vinegar and analyze their nutritional components.The results of the extraction and analysis show that36volatile compounds were identified (match rate at80%and above). These compounds were classified into7categories including5esters,5acids,4ketones, 10alcohols,6aldehydes,2phenols and4paraffins. Amino-acid Analysis was adopted to detect the existence of free amino acids in the whey vinegar. The result reveals the existence of25types of amino acids. Eight (8) types of these amino acids are essential to the health of humans and4types, including lysine, threonine, arginine, serine, et cetera, can enhance the immune system of humans.A test was performed on the16mineral and vitamin contents in the whey vinegar. The result shows that whey vinegar contains a lower level of Chlorine, Sodium and Iron than those in white rice vinegar and apple cider vinegar, but contains a higher level of potassium, calcium, magnesium, zinc, manganese, iodine, phosphorus, selenium, VB1, VB2, Vc, VA, and VE than Those in white rice vinegar and apple cider vinegar.3. An animal laboratory test was conducted to see if whey vinegar could improve the immune system of a common rat. the results are as follows:(1) The ratio of internal organ to body weight test:Compared with control groups, the dosage feeding groups (experimental groups) at high and mid-levels of feeding show statistically significant enhancement of weight in immune organs and statistically significant enhancement of thymus and spleen indexes (P<0.05).(2) The test of delayed hypersensitivity in rats induced by dinitrofluorobenzene: Compared with control groups, the dosage feeding groups (experimental groups) at high, mid-and low levels of feeding show statistically significant differences in ear swelling of rats (P<0.05), an indicator that whey vinegar can significantly increase delayed hypersensitivity in rats.(3) The test of lymphocyte transformation induced by ConA:Compared with control groups, the dosage feeding groups (experimental groups) at high, mid-and low levels of feeding show statistically significant enhancement in lymphocyte proliferation (P<0.01).(4) The test of antibody-forming cells:Compared with control groups:the dosage-feeding groups (experimental groups) at high, mid-and low levels of feeding show that whey vinegar has no statistically significant effect on antibody forming cells.(5) The test of serum hemolysin:Compared with control groups, the dosage feeding groups (experimental groups) at high, mid-and low levels of feeding show that whey vinegar has no statistically significant effect on serum hemolysin, or on antibody formation level.(6) The test of phagocytosis of chicken red blood cells in mice peritoneal macrophages:Compared with control groups, the dosage feeding groups (experimental groups) at high and mid-levels of feeding show that whey vinegar can effect statistically significant enhancement of rats’macrophage phagocytosis ability to attack and swallow (P＜0.01).(7) The test of carbon clearance:Compared with control groups, the dosage feeding groups (experimental groups) at high, mid-and low levels of feeding show that whey vinegar can effect statistically significant increases in rats’swallow indexes (P＜0.05).(8) The determination of the activity of NK cells:Compared with control groups, the dosage feeding groups (experimental groups) at high, mid-and low levels of feeding show that whey vinegar can effect statistically significant enhancement of the activity of NK cells in rats(P＜0.01).In conclusion, whey vinegar can be successfully produced from milk whey, with liquid state fermentation process at the alcoholic and acetic acid stages. Whey vinegar contains a higher level of nutrients than a regular white rice or apple cider vinegar. In addition, whey vinegar is rich in flavor. Finally, whey vinegar can significantly improve a rat’s immunity.更多还原