乳铁蛋白的分离及纯化

乳铁蛋白 (Lactoferrin ,简称Lf)作为一种糖蛋白 ,主要存在于哺乳动物的各种外分泌物中 ,如乳汁、眼泪等 ,乳汁尤其是初乳中Lf的含量很高[1] 。Lf是一种多功能蛋白质 ,不仅具有广谱的抗菌作用 ,而且还能够增强机体的抗病毒、抗感染、抗肿瘤的能力和免疫能力。因此 ,从来源丰富的牛初乳中提取出Lf,把它作为功能性成分应用于食品行业 ,具有广阔的应用和开发前景。本文通过盐析法和层析法从牛初乳中分离出乳铁蛋白 ,并应用酶联免疫法 (ELISA)检测不同分离纯化方法所获得的乳铁蛋白的含量 ,并用电泳分析其纯度。所获得的乳铁蛋白的分子质量为 780 0 0u ,其纯度大约为 92 %。     

牛初乳中乳铁蛋白的分离及纯化

乳铁蛋白(Lactoferrin,简称Lf)作为一种糖蛋白,主要存在于哺乳动物的各种外分泌物中,如乳汁、眼泪、唾液等,乳汁尤其是初乳中Lf的含量很高犤1犦。本文通过盐析法和有机溶剂沉淀法从牛初乳中分离出乳铁蛋白,并应用酶联免疫法鉴定各步骤中分离的乳铁蛋白,获得的乳铁蛋白的相对分子量为75,000Da。     

乳铁蛋白的研究进展

综述了乳铁蛋白最新研究成果,主要包括乳铁蛋白分子结构、生物学功能及重组乳铁蛋白的研究现状。乳铁蛋白不仅具有广谱的抗菌性质,而且还能够增强机体的抗病毒、免疫能力,促进肠道健康。作为一种新型的具有多种功能的蛋白质,有望在食品药品中取得更广泛的用途。     

乳铁蛋白理化性质及其成骨作用研究进展

乳铁蛋白是一种具有抗菌、免疫调节等多功能性的铁结合糖蛋白,主要存在于哺乳动物乳汁中。近年来,乳铁蛋白在促进骨生长方面的功能逐渐成为研究热点。本文主要综述乳铁蛋白的结构、理化性质及其对成骨细胞、破骨细胞的作用、骨生长的影响及机制等方面的研究进展,并展望乳铁蛋白作为骨效应分子的应用前景及仍需解决的问题。     

乳铁蛋白与纳米脂质体的相互作用及对磷脂膜结构的影响

为探明纳米脂质体(N-lip)与食品蛋白的相互作用，采用薄膜分散法制备N-lip。以乳铁蛋白(Lf)为模型蛋白，评价不同蛋白浓度和滴加方式下形成的纳米脂质体/乳铁蛋白(N-lip/Lf)复合物的粒径、Zeta-电位和多分散系数，并用隐形率(Fs)来估计N-lip与Lf的结合程度。利用透射电子显微镜比较乳铁蛋白作用前、后的形貌变化，采用光谱法和X射线衍射分析Lf与N-lip的相互作用。结果表明:在磷脂质量浓度为10.0 mg/mL、磷脂与胆固醇质量比为10 ∶ 1、磷脂与Tween-80的质量比为4 ∶ 1、蛋白质量浓度为0.1 mg/mL时，形成的N-lip/Lf复合物平均粒径为(100.80±0.141) nm，电位为(-54.9±2.276) mV，具有较好的分散度。透射电子显微镜观察可见，Lf在N-lip边缘形成浅色包裹层。Lf与N-lip的相互作用促进了Lf中色氨酸和酪氨酸残基所处环境的疏水性增强，Lf的α-螺旋结构含量增加；Lf可以通过氢键、范德华力和疏水作用与N-lip磷脂双分子层膜发生相互作用，吸附到磷脂膜表面。研究结果可为脂质体与食品蛋白的互作及脂质体在食品工业中的潜在应用提供理论依据。     

乳铁蛋白对卵清白蛋白过敏小鼠外周血中Th1/Th2细胞平衡的影响

利用卵清白蛋白(OVA)建立BALB/C小鼠食物过敏模型,观察乳铁蛋白(lactoferrin,Lf)对该食物过敏机体的免疫调节作用。小鼠分别接受预防性乳铁蛋白处理和胃肠道刺激期间灌胃乳铁蛋白,最后一次处理1h后检测其外周血中IFN-γ、IL-4、TGF-β、IL-6、IL-10、IL-12p40和IgE的变化。结果表明:2mg/(kg.d)(以体质量计)的Lf在胃肠道刺激期间显著提高了IFN-γ和IL-12p40水平而降低了IL-4和IL-6水平,但20mg/(kg.d)的Lf降低了所有这些细胞因子。同时,Lf明显降低了血清中IgE浓度而提高了IFN-γ/IL-4比值,20mg/(kg.d)的Lf对IgE影响较大,2mg/kg的Lf对IFN-γ/IL-4比值影响较大。此外,预防性处理中的Lf均显著降低了IL-4、IL-6、IL-10、IL-12p40、IFN-γ、TGF-β和IgE水平,并且20mg/(kg.d)的Lf效果较好,且该剂量明显提高了IFN-γ/IL-4比值。因此,Lf能够改善OVA诱导的食物过敏小鼠Th1/Th2细胞的平衡,具有重要的免疫调节作用。     

乳铁蛋白的开发利用及其研究进展

综述了乳铁蛋白的基本性质、功能特性及其一些最新的研究进展，并对牛初乳中乳铁蛋白的开发利用进行了展望。     

三元公司乳品新技术研究最新进展——人乳铁蛋白基因及真菌中表达(连载四)

乳铁蛋白作为一种活性蛋白，在功能营养食品及医药等方面具有广阔的应用前景。但很久以来，如何获得廉价的人乳铁蛋白一直是开发乳铁蛋白遇到的最大难题，而利用基因工程技术生产重组蛋白给乳铁蛋白的开发利用提供了可行性途径，这也是乳铁蛋白研究的发展趋势。随着基因工程技术和遗传学的迅速发展，以及人们对乳铁蛋白基因及其表达调控机制深人了解，重组生产将成为的人乳铁蛋白来源的主要渠道。     

乳及乳制品中乳铁蛋白含量的电泳迁移率检测

凝胶电泳迁移率(EMSA)是研究蛋白质和DNA分子相互作用的经典分子生物学方法。本研究根据乳铁蛋白(Lactoferrin,Lf)和DNA分子的特异性结合,基于Lf-DNA复合物和DNA分子迁移率差异,建立了一种聚丙烯酰胺凝胶电泳(PAGE)检测乳铁蛋白的方法。实验中以FAM荧光基团标记DNA分子,根据凝胶成像图中Lf-DNA复合物荧光条带的存在与否、荧光条带灰度值的大小,来判断样品中乳铁蛋白是否存在、含量多少,从而实现乳铁蛋白的定性与定量检测。结果表明,该方法具有良好的特异性及准确性,经过简单稀释后便可直接应用于乳及乳制品中乳铁蛋白的准确定量检测。     

乳铁蛋白与肿瘤关系的研究进展

乳铁蛋白是一种天然糖蛋白,主要存在于母乳中,具有多种生物学功能。近年来,一些研究认为,乳铁蛋白具有化学预防和抗肿瘤作用。本文就乳铁蛋白和肿瘤的关系作一综述。     

浅谈乳铁蛋白在肉类保鲜中的应用

乳铁蛋白作为一种天然的、具有许多特殊生理功能的糖蛋白,已经引起国际上广泛关注。本文叙述了乳铁蛋白的抗菌机制,包括:乳铁蛋白的结构特性、抗菌作用、抗菌机制等,并对其在肉类保鲜中的应用前景作了展望。     

乳铁蛋白及其生理功能

乳铁蛋白是一种天然糖蛋白,主要存在于哺乳动物各种外分泌物中。因乳铁蛋白在抑菌、抗病毒、免疫调节、促进铁吸收等方面具有独特生理功能,从而对乳铁蛋白开发和应用具有广阔前景。     

Lactoferrin research,technology and applications

Lactoferrin is an iron-binding glycoprotein present in milk as well as other exocrine secretions and neutrophil granules in mammals. Lactoferrin is considered to be an important host defense molecule and has a diverse range of physiological functions such as antimicrobial/antiviral activities, immunomodulatory activity, and antioxidant activity. During the past decade, it has become evident that oral administration of lactoferrin exerts several beneficial effects on the health of humans and animals, including anti-infective, anticancer, and anti-inflammatory effects. This has enlarged the application potential of lactoferrin as a food additive. The technology of producing bovine lactoferrin on a factory scale was established over 20 years ago. Bovine lactoferrin is purified by cation-exchange chromatography from bovine skim milk or whey, and is commercially available from several suppliers. Recombinant human lactoferrin is produced by Aspergillus niger, transgenic cows, and rice, and its efficacy is being evaluated. In this article, we review basic research and technological aspects of the application of lactoferrin.     

乳铁蛋白的抑菌作用及其机制

叙述了乳铁蛋白和乳铁蛋白多肽的抗菌机制。包括:乳铁蛋白的结构特性、抗病毒、抗菌功能、抗菌机制、影响抑菌活性的因素等,并对其应用前景作了展望。     

乳铁蛋白的国内外研究动态及趋势

乳铁蛋白是一种铁结合性糖蛋白，具有多种生物学功能，近年来成为国内外研究的热点。本文综述了乳铁蛋白的近期研究热点：活性乳铁蛋白的开发应用，高效表达人乳铁蛋白的转基因技术进展，工业化生产途径的最新研究以及乳铁蛋白的开发应用前景。     

乳铁蛋白最新研究进展--结构、特性及生产制备方法(Ⅰ)

乳铁蛋白广泛存在于哺乳动物的分泌物中,如人乳或牛乳中,具有较多的生理活性功能.文章综述了乳铁蛋白的一些最新研究进展包括分子结构、一些功能特性以及生产制备方法.     

The effect of bovine milk lactoferrin on human breast cancer cell lines

The evidence that biologically active food components are key environmental factors affecting the incidence of many chronic diseases is overwhelming. However, the full extent of such components in our diet is unknown, as is our understanding of their mechanisms of action. Beyond the interaction of these food components with the gut and intestinal immune functions, whey proteins such as lactoferrin are being tested as anticancer agents. Lactoferrin is an iron-binding protein that has been reported to inhibit several types of cancer. In the present work, the effects of bovine milk lactoferrin on human breast cancer HS578T and T47D cells were studied. The cells were either untreated or treated with lactoferrin concentrations ranging from 0.125 to 125 μM. Lactoferrin decreased the cell viability of HS578T and T47D by 47 and 54%, respectively, and increased apoptosis about 2-fold for both cell lines. Proliferation rates decreased by 40.3 and 63.9% for HS578T and T47D, respectively. For the T47D line, cell migration decreased in the presence of the protein. Although the mechanisms of action are not fully known, the results gathered in this work suggest that lactoferrin interferes with some of the most important steps involved in cancer development.     

乳铁蛋白的特性、分离纯化方法及特点

乳铁蛋白是一种铁结合性糖蛋白,在初乳中含量较高,乳铁蛋白具有多种生物学功能,如抗菌性、抗氧化、抗病毒、抗癌症等。乳铁蛋白的分离纯化方法较多,就吸附色谱法、离子交换色谱法、亲和色谱法、固定化单克隆抗体法、超滤法、盐析法等进行了概述,并就其工业化生产方法进行了展望。     

Osteogenic effects of the peptide fraction derived from pepsin-hydrolyzed bovine lactoferrin

Osteoporosis is a common disease that frequently occurs in the older population, particularly in postmenopausal women. It severely compromises the health of the older population, and the drugs commonly used to treat osteoporosis have a variety of adverse effects. Lactoferrin (LF) is a protein present in milk that has recently been found to exhibit osteogenic activity. Lactoferrin is nontoxic and harmless, suggesting that it may have excellent biocompatibility and tolerability after human consumption. Oral consumption of LF in an ovariectomized rat model has been found to ameliorate osteoporosis. However, the mechanism underlying this effect remains to be clarified. In this study, bovine LF (bLF) was first hydrolyzed by pepsin for 1 h, and the hydrolyzed mixture was freeze-dried and collected. The hydrolyzed mixture was then separated into 5 components (E1–E5), of which E3 had the greatest effect in promoting proliferation of osteoblasts (MC3T3-E1). Component E3 was further isolated into 21 components with preparative reversed phase HPLC, and the E3-15 component had maximal bioactivity. With HPLC-mass spectrometry and peptide sequencing, E3-15 was identified to contain amino acids 97 to 208 from the bLF N terminus. Then, E3-15 was divided into 6 different peptide segments (P1–P6), and the corresponding segments were generated by solid-phase synthesis. Only the P1 peptide (amino acids 97–122 from the N terminus of bLF) significantly promoted osteoblast proliferation. The bioactivity of P1 toward osteoblast cells and alkaline phosphatase activity were tested as a function of P1 concentration, and a nonlinear effect was observed.