羊乳乳清蛋白组成和功能及其与人乳、牛乳的对比分析

本研究将羊乳、牛乳、人乳中乳清蛋白进行分离并结合液质联用技术鉴定,在羊乳、牛乳、人乳乳清蛋白中分别鉴定出156、278、454种蛋白质。与牛乳与人乳乳清蛋白对比显示,羊乳含有99种特异性表达蛋白质,与牛乳和人乳分别有31种和15种相同表达蛋白质。通过分析基因本体(gene ontology,GO)功能注释发现,羊乳乳清蛋白在生物过程中主要发挥生物调节作用;在分子功能上,主要体现在结合作用方面;在细胞组成上,参与的细胞组成主要为细胞器区和胞外区。羊乳乳清蛋白在以上三种功能上与人乳有较大差距,但与牛乳相近。通过分析京都基因与基因组百科全书系统(Kyoto Encyclopedia of Genes and Genomes,KEGG)代谢通路可知,羊乳主要参与补体和凝血级联反应以及吞噬作用,对人体免疫能力有积极影响。对羊乳与人乳、牛乳乳清蛋白组成及功能区别的研究,为羊乳的进一步研究和开发提供一定的理论参考。     

A Simple and Fast Detection Method for Bovine Milk Residues in Foods: A 2‐Site Monoclonal Antibody Immunochromatography Assay

The ingredient declaration on food labels assumes paramount importance in the protection of food‐allergic consumers. China has not implemented Food allergen labeling. A gold immunochromatography assay (GICA) was developed using 2 monoclonal antibodies (mAb) against the milk allergen β‐lactoglobulin in this study. The GICA was specific for pure milk samples with a sensitivity of 0.2 ng/mL. Milk protein traces extracted from 110 food products were detected by this method. The labels of 106 were confirmed by our GICA method: 57 food samples originally labeled as containing milk were positive for β‐lactoglobulin and 49 food samples labeled as not containing milk were negative for β‐lactoglobulin. However, 3 food samples falsely labeled as containing milk were found to contain no β‐lactoglobulin whereas 1 food sample labeled as not containing milk actually contained β‐lactoglobulin. First, these negatives could be because of the addition of a casein fraction. Second, some countries demand that food manufacturers label all ingredients derived from milk as “containing milk” even though the ingredients contain no detectable milk protein by any method. Our GICA method could thus provide a fast and simple method for semiquantitatation of β‐lactoglobulin in foods. Practical Application: The present method provides a fast, simple, semiquantitative method for the determination of milk allergens in foods.     

转糖基β-半乳糖苷酶生产含低聚半乳糖的低乳糖牛奶

研究利用具有转糖基活性的β-半乳糖苷酶生产含低聚半乳糖的低乳糖牛奶。含低聚半乳糖的低乳糖牛奶既能解决乳糖不耐症问题，同时还在牛奶中增添了低聚半乳糖双歧因子。从Enterobacter sp．B5的无细胞抽提液中，利用硫酸铵沉淀制备具有转糖基活性的β-半乳糖苷酶，作用鲜牛奶生成含低聚半乳糖的低乳糖牛奶。利用薄层层析、高压液相色谱技术和软件NIHImageJ1．36分析了反应产物中的糖组分（葡萄糖、半乳糖、乳糖和低聚糖（包括转移二糖和三糖））。结果显示酶作用后的鲜牛奶中大约70％的乳糖转化为葡萄糖和半乳糖，至少10％的乳糖通过转糖基反应生成低聚半乳糖。在酶浓度为3U／ml，50℃酶解4h的反应条件下，获得含低聚半乳糖的低乳糖牛奶的糖组成为0．59％低聚半乳糖（含0．37％转移二糖和0．22％三糖），1．33％乳糖，2．83％单糖。本研究利用软件NIHImageJ1．36定量分析样品糖组分的TLC结果，可以避免乳糖的同分异构体．转移二糖在高压液相色谱结果中因无法分离而造成的漏检。     

母乳中的低聚半乳糖研究和应用的最新进展

本详述了低聚半乳糖的研究和应用的最新进展,并简述了低聚半乳糖的特性。     

云南撒尼地区乳饼及酸乳清中优良乳酸菌的筛选和发酵性能研究

选取SDC4-1、MDC3-1、WDC3-1 3株从云南撒尼地区乳饼和酸乳清中分离鉴定的乳酸菌株,用耐酸、耐胆盐实验筛选其中的优势菌株;并用产酸能力、后酸化、产乙醛能力、蛋白水解酶测定和β-半乳糖苷酶测定等实验研究优势菌株的发酵性能。3株菌均能耐酸,其中MDC3-1耐酸能力最高,达254%,WDC4-1最低,为52%,但仅SDC4-1可耐受胆盐,在0.5%的胆盐中,4 h后的存活率为81%,SDC4-1是优势菌。SDC4-1发酵性能实验中,SDC4-1菌株具有很好的凝乳状态,发酵酸奶口感风味浓郁,在贮藏过程中活菌数含量高,冷藏15 d后活菌数达到了9.79 lg CFU/g,远超国家标准。产酸较快,13 h时,酸度已达82.0°T,后酸化性好,产乙醛能力较好,蛋白水解能力强,产β-半乳糖苷酶的量高,达7.7μg/mol,能保证酸奶在冷藏过程中品质稳定,因而筛选出具有优良发酵特性的菌株SDC4-1可以为发酵食品提供更广泛的素材。     

不同泌乳期羊乳和牛乳的高通量定量乳清蛋白质组学

利用高分辨质谱技术获取牛初乳、牛常乳、羊初乳和羊常乳的乳清蛋白组轮廓,基于其非标记定量强度建立偏最小二乘判别分析模型。结果发现牛初乳和牛常乳在蛋白质组成方面较羊初乳和羊常乳更相似,同时筛选出羊乳中丰度较高的9 种蛋白用作标志物区分这4 组乳样。生物信息学分析发现羊乳中的高丰度蛋白大部分与免疫应答和代谢过程有关,说明羊乳更有助于新生儿建立抗微生物感染的免疫系统。该研究可加深对羊乳蛋白的认识,对牛乳及其制品的营养改良和母乳替代品的生产具有重要意义。     

Preparation of lactose‐free milk by using salt‐fractionated almond (Amygdalus communis) β‐galactosidase

β‐galactosidase was isolated from almond (Amygdalus communis) extract by ammonium sulfate precipitation. Almond proteins precipitated by using ammonium sulfate and then dialysed exhibited 5.3‐fold purification of β‐galactosidase, and the yield of enzyme preparation was 96.5%. The partially purified β‐galactosidase exhibited pH and temperature optima at pH 5.5 and 50 °C, respectively. The enzyme was significantly stable against heat, pH, calcium and magnesium ions and D ‐galactose. The almond β‐galactosidase preparation exhibited over 89% activity even after 2 months storage at 4 °C. Hydrolysis of lactose in milk and whey was performed in a stirred batch process by using this enzyme preparation. These observations indicated that the hydrolysis of lactose increased continuously with time. The enzyme could hydrolyse 94% of lactose in buffer solution and whey whereas 90% of lactose hydrolysis was achieved in milk. The main aim of the present study was to prepare lactose‐free milk, which must be free from contamination, and the process should be inexpensive. Copyright © 2007 Society of Chemical Industry     

牛初乳和牛常乳乳清N-糖蛋白质的差异分析

为阐明牛乳蛋白质N-糖基化,采用糖蛋白质组学技术,在牛初乳和牛常乳乳清中共鉴定到154 个N-糖蛋白和246 个糖基化位点,其中,牛初乳鉴定到117 个N-糖蛋白和183 个糖基化位点;牛常乳鉴定到109 个N-糖蛋白和145 个糖基化位点。初乳中丛生蛋白（P17697）糖基化位点N-283表达量最高,常乳中α-乳白蛋白糖基化位点N-93表达量最高。考虑定量差异及有无差异,在牛初乳和牛常乳乳清中共鉴定到129 个糖蛋白的190 个差异表达糖基化位点。基因本体论功能注释表明,差异表达糖蛋白参与的生物学过程是生物调节、刺激性反应、多细胞生物过程、定位、免疫系统过程等;主要分布为细胞外区域和细胞器;主要的分子功能是结合作用、催化活性和分子功能调节。差异表达糖蛋白参与的代谢通路主要是补体与凝血级联、金黄色葡萄球菌感染和溶酶体等。此外,通过蛋白互作分析,找到一些具有高连接度的重要糖蛋白。本研究丰富了牛乳N-糖蛋白质组成及其糖基化位点信息,阐明了牛乳乳清N-糖基化的功能,为评价和改善牛乳品质、研发婴幼儿配方乳中糖蛋白质的改良及功能食品的生产提供了理论依据。     

Simple and Rapid Method for Measuring Turbidity in Gels and Sols from Milk Whey Protein

Protein isolates prepared from acid whey concentrate and cheese whey concentrate, and β-lactoglobulin were studied. Turbidity of samples was measured using a 96-well microplate and a microreader. This method allowed many (>100) small samples (< 250 μL) to be treated at the same time. At low ionic strength and at neutral to alkaline pH, samples were transparent after heating. Transparent gels could also be prepared in this region with a small amount of NaCl.     

Purification and physicochemical characterization of ovine β‐lactoglobulin and α‐lactalbumin

Ovine whey proteins were fractionated and studied by using different analytical techniques. Anion‐exchange chromatography and reversed‐phase high‐performance liquid chromatography (HPLC) showed the presence of two fractions of β‐lactoglobulin but only one of α‐lactalbumin. Gel permeation and sodium dodecyl sulfate (SDS)‐polyacrylamide gel electrophoresis allowed the calculation of the apparent molecular mass of each component, while HPLC coupled to electrospray ionisation‐mass spectrometry (ESI‐MS) technique, giving the exact molecular masses, demonstrated the presence of two variants A and B of ovine β‐lactoglobulin. Amino acid compositions of the two variants of β‐lactoglobulin differed only in their His and Tyr contents. Circular dichroism spectroscopy profiles showed pH conformation changes of each component. The thermograms of the different whey protein components showed a higher heat resistance of β‐lactoglobulin A compared to β‐lactoglobulin B at pH 2, and indicated high instability of ovine α‐lactalbumin at this pH.     

Production of soya milk containing low flatulence‐causing oligosaccharides in a packed bed reactor using immobilised α‐galactosidase

Peanut α‐galactosidase was immobilised in calcium alginate beads and used to hydrolyse the flatulence‐causing oligosaccharides, raffinose and stachyose, in soya milk in batch and in packed bed reactor with recycle. The immobilised enzyme exhibited a slightly lower activity than the free enzyme. The activity yield of immobilised α‐galactosidase was 75.1% and the immobilisation yield was 82.6%. Batch hydrolysis using immobilised enzyme at 55 °C resulted in 96% reduction in the oligosaccharides after 12 h. For the continuous process, a packed bed reactor with recycle was used. More than 98% of the oligosaccharides were hydrolysed after 6 h of reaction at 55 °C. The immobilised enzyme also proved to be stable up to three repeated hydrolysis reactions.     

乳清蛋白及与其它乳成分的热聚合作用机制研究进展

乳蛋白决定奶类品质,而热加工会影响乳清蛋白尤其是β-乳球蛋白的稳定性。热处理时,乳清蛋白不仅自身发生不同程度的聚合,而且通过巯基-二硫键分别与酪蛋白胶束和乳脂球膜蛋白发生结合。乳清蛋白亦可与其它乳成分如乳糖、钙盐、乳脂发生热聚合作用。本文根据乳品受热温度的不同,针对乳清蛋白间及其与其它乳成分的相互作用途径进行分析,阐明热聚合作用过程及机理,对改善乳制品的热稳定性、凝胶性等功能性质具有重要的理论意义,拓宽乳清蛋白作为配料在相关食品体系中的应用。     

乳清蛋白在食品工业中的应用

对乳清蛋白在食品中的应用进行了简要的介绍,其中对乳清蛋白的主要组分(α- 乳白蛋白、β- 乳球蛋白、乳铁蛋白等)、功能特性(成胶性,涂层性和成膜性等)、生产技术(膜分离技术、吸附分离法、亲和色谱提纯法等)及改性方法(物理改性及酶改性)做了详细的介绍,并对我国乳清蛋白的应用前景进行了展望。     

Multiple forms of β‐galactosidase from mango (Mangifera indica L Alphonso) fruit pulp

Mango (Mangifera indica L cv Alphonso) was found to contain three isoforms (I, II and III) of β‐galactosidase which, upon purification on Sephadex G‐200, had relative abundances of 44, 38 and 18%, respectively. The total specific activity increased from 20 to 727 µmol l^?1 upon purification, representing a ～36‐fold increase with a recovery of 0.28 U U^?1. The optimal pH for activity and stability were in the ranges 3.6–4.3 and 4–6.2, respectively. The optimal temperature for β‐galactosidase activity was between 42 and 47 °C with T_m in the range 45–51 °C. The K_m for pNP‐β‐galactopyranoside was 0.98, 1.11 and 0.95 mM , and V_max was 0.56, 0.53 and 0.35 µmol pNP min^?1, respectively for isoforms I, II and III. Hg²⁺ caused strong inhibition, whereas galacturonic acid, galactose, xylose, fucose and mannose slightly inhibited the activity of β‐galactosidase isoforms. The apparent molecular weights by GPC were 78, 58 and 91 kDa for isoforms I, II and III, respectively. The ability of these isoforms to degrade the endogenous substrate (arabinogalactan) possibly suggests a role in pectin dissolution during tissue softening/fruit ripening. Copyright © 2004 Society of Chemical Industry     

关于免疫乳与初乳

本文综述了免疫乳和初乳的发展、制备和作用,分析了免疫乳和初乳存在的问题,提出免疫乳与牛初乳只能作为增强肠道局部免疫的保健食品,但对预防肠道传染病的时效性提出质疑,并指出长期服用可能会产生抗抗体的论点。     

牛乳重要营养品质特征的研究进展

牛乳营养组成非常丰富,乳蛋白和乳脂肪是表征牛奶营养品质的重要指标,并受诸多因素的影响。本文综述遗传、环境和饲养管理等因素对乳蛋白和乳脂肪组分及含量的影响,以及牛乳蛋白和乳脂肪酸的特征性含量和比值。