婴幼儿乳粉中生物素含量的测定研究

研究利用微生物法测定婴幼儿乳粉中生物素的含量。利用植物乳杆菌(ATCC 8014)对生物素具有很高灵敏性的特点,其生长与生物素含量成一定的线性关系,通过测定培养后菌液吸光度的变化得出婴幼儿乳粉中生物素的含量。测得的标准曲线在0.1ng～1ng范围内线性关系良好(R2=0.9989),所测样品RSD为2.8%(n=6),标准品平均回收率为89.3%～97.7%。该方法重现性较好,稳定可靠,适用于婴幼儿乳粉中生物素含量的测定。     

乳杆菌体外降解胆固醇特性研究

以实验室自主分离纯化的鼠李糖乳杆菌zxr01、嗜酸乳杆菌zrx02、植物乳杆菌zrx03, 3株乳杆菌体外降解胆固醇的特性以及不同金属阳离子对其影响进行了研究。嗜酸乳杆菌、鼠李糖乳杆菌、植物乳杆菌体外胆固醇降解率分别为28.87%、25.61%、20.39%；当pH为3时，3株乳杆菌耐酸性分别为：52.77%、52.57%、59.82%;牛胆盐浓度为0.3%时胆盐耐受性为28.65%、27.40%、26.04%；嗜酸乳杆菌在氯仿中的疏水性最好，达到了19.88%，静置24 h后，嗜酸乳杆菌和植物乳杆菌的自凝聚力都达到了80%以上且3株乳杆菌产生白色沉淀说明能产生胆盐水解酶，在Ca2+浓度为0.02 N时，嗜酸乳杆菌的胆固醇降解率为35.4%和降解效果65.43μg/u。结果显示3株乳杆菌均能在体外降解胆固醇，嗜酸乳杆菌的降解胆固醇的能力要优于其它两株乳杆菌，添加浓度为0.02 N的Ca2+可以促进嗜酸乳杆菌胆固醇降解率和降解效力。本研究结果可为进一步研究胆固醇降解提供了试验依据，为开发降胆固醇食品提供材料。     

微生物法检测叶酸、生物素、维生素B12接种液制备方法的优化

目的解决使用微生物法GB 5009.211-2014、GB 5009.259-2016、GB 5413.14-2010检测食品中叶酸、生物素、维生素B_(12)接种液制备耗时长的问题。方法将标准方法制备的接种液与甘油水按1:1(V:V)比例混匀,分装于小离心管,-80℃冻存。再通过用冻存不同时间的接种液检测样品,分析标准曲线线性、中间精密度和加标回收率来验证制备方法的适用性。结果标准曲线线性关系r~2均0.99,中间精密度叶酸RSD为1.5%～3.2%,生物素RSD为1.7%～4.2%,维生素B_(12) RSD为2.5%～9.3%;回收率方面,叶酸为99.5%～101.0%,生物素98.6%～101.3%,维生素B_(12)90.3%～96.4%。鼠李糖乳杆菌、植物乳杆菌、莱氏曼氏乳杆菌接种液超低温冻存有效时间长分别为6、8、3个月。结论优化后的接种液制备方法缩短了检测周期,操作简便、方法可靠,适合检测使用。     

银耳多糖对植物乳杆菌增殖作用的研究

文章以植物乳杆菌为研究对象,通过测定植物乳杆菌利用银耳多糖的生长曲线、电导率、活菌数统计及菌体表面形态的变化,用DPS软件进行数据统计与分析来研究银耳多糖对植物乳杆菌增殖作用的影响,并确定银耳多糖是否能作为植物乳杆菌的益生元。结果表明,银耳多糖对植物乳杆菌有益生作用,能够作为植物乳杆菌的益生元,对其菌体完整性无损坏并无毒害作用,且试验测得植物乳杆菌利用银耳多糖的最适浓度为0.20%。     

乳酸菌发酵蔬菜汁工艺研究

本文以胡萝卜、番茄为原料，经过加工处理，将所得的蔬菜汁用嗜酸乳杆菌和植物乳杆菌进行发酵，确定了蔬菜汁发酵的最佳比例为番茄汁：胡萝卜汁为3：1， 蔬菜汁含量为60%，确定了蔬菜汁发酵的最佳工艺参数为发酵温度32℃，嗜酸乳杆菌、植物乳杆菌接种量分别为5%、1%，发酵时间为12h。     

植物乳杆菌微胶囊化研究

为保护植物乳杆菌的活性以增强乳杆菌在动物肠道内的益生功能,以天然发酵玉米青贮饲料中优良植物乳杆菌作为芯材,乳清蛋白和明胶为壁材,利用喷雾干燥法制成微胶囊,并以植物乳杆菌包埋率为响应值,研究壁材配比、壁材添加量、进风温度、进料量4个因素,进行中心组合实验(Box-Behnken),通过响应面分析对喷雾干燥法制备植物乳杆菌微胶囊条件进行优化。结果表明:最优条件为壁材配比(乳清蛋白与明胶质量比)1:2、壁材添加量22%、进风温度127℃、进料量35%,在此条件下,植物乳杆菌包埋率为62.15%。结论:本研究为应用喷雾干燥法制备植物乳杆菌微胶囊奠定了基础。     

可降胆固醇的优势乳杆菌筛选

为丰富降胆固醇、降血糖的益生菌资源,以实验室10株潜力益生菌株为实验对象,进行体外降胆固醇能力、胆盐水解酶(BSH)活性及α-葡萄糖苷酶抑制率试验,测定菌株对人工胃液和胆盐的耐受性,评价优势菌株的细胞黏附性能及对抗生素的耐药安全性能。结果表明,植物乳杆菌LH-511、植物乳杆菌10-12、植物乳杆菌10-4对胆固醇的降解率在50%以上,显著高于商业菌株植物乳杆菌299V(p<0.05)。植物乳杆菌10-12、植物乳杆菌SD-H9对α-葡萄糖苷酶的抑制率高达41.7%、40.1%。植物乳杆菌LH-511、植物乳杆菌10-12、植物乳杆菌10-4、植物乳杆菌10-14、卷曲乳杆菌OF48-2pH5这5株菌具有较好的BSH活力、α-葡萄糖苷酶抑制性、抗逆性以及对HT-29细胞的黏附能力。但仅植物乳杆菌LH-511和卷曲乳杆菌OF48-2pH5通过了10种抗生素的安全性试验。综上所述,植物乳杆菌LH-511和卷曲乳杆菌OF48-2pH5具有较好的降胆固醇、降血糖潜力,且通过了抗逆性、黏附性、安全性试验,可用于进一步的开发和应用。     

传统发酵牦牛乳中2株高产胞外多糖乳酸菌在模拟消化道中耐受力的研究

从传统发酵牦牛发酵乳中分离出的2株高产胞外多糖乳酸菌,为了研究其在模拟消化道中耐受力,对2株乳酸菌(植物乳杆菌、干酪乳杆菌)分别在人工胃液、人工肠液、人工胆汁和高盐4个模拟人工胃肠道消化环境中进行培养,测其耐受力以及对Caco﹣2细胞的黏附能力。结果表明:植物乳杆菌和干酪乳杆菌在人工胃液中作用3 h的存活率随pH值的增大而增加。在pH4.5时,植物乳杆菌的存活率达到53.63%,干酪乳杆菌的存活率达到50.83%;在人工肠液中作用4h,植物乳杆菌的存活率达到了59.58%,干酪乳杆菌的存活率达到了51.42%;在胆盐环境中培养24 h后的植物乳杆菌和干酪乳杆菌活菌数随牛胆盐质量浓度的增加而降低,活菌数均保持在108cfu/m L以上;在高盐环境中培养24 h后的活菌数随盐质量浓度的增加而降低,活菌数均在108 cfu/mL以上;并且2株乳酸菌的黏附能力也很强,植物乳杆菌可以达到16.83%、干酪乳杆菌可以达到14.86%。结论:植物乳杆菌和干酪乳杆菌均能通过胃进入肠道并保持活力,而且能在肠道很好地定植,为植物乳杆菌和干酪乳杆菌作为益生菌应用在食品中提供了理论基础。     

辣木叶多糖对益生菌生长及耐受性的影响

利用体外发酵试验考察不同质量浓度(0.2%、0.5%、1%、2%、4%)辣木叶多糖对保加利亚乳杆菌(Lactobacillus bulgaricus)和植物乳杆菌(Lactobacillus plantarum)增殖作用及在模拟人体胃肠道逆环境中的耐受能力的影响。研究结果表明,辣木叶多糖可显著促进保加利亚乳杆菌和植物乳杆菌的生长。随着辣木叶多糖含量的增加,益生菌液浓度先增加后降低,在多糖质量浓度为2%时菌液浓度达到最大,表明辣木叶多糖添加量为2%时最有利于益生菌的生长。辣木叶多糖可显著提高保加利亚乳杆菌和植物乳杆菌对酸和胆汁盐的耐受性(P0.05),可显著增强保加利亚乳杆菌对胃肠液的耐受性(P0.05),但对植物乳杆菌肠液的耐受性影响不显著。     

低聚半乳糖对植物乳杆菌发酵乳特性及抗蜡样芽孢杆菌活性的改善

为探究低聚半乳糖对植物乳杆菌发酵乳特性及抗菌活性的影响,本文采用单因素法考察影响发酵乳特性的主要因素,并以响应面法优化发酵乳最佳发酵条件;以产肠毒素蜡样芽孢杆菌HN001为指示菌,探究低聚半乳糖的添加对植物乳杆菌ZDY2013发酵乳抑菌活性的改善作用。结果表明:植物乳杆菌能有效利用低聚半乳糖进行体外代谢,并抑制蜡样芽孢杆菌生长;牛奶中添加适量低聚半乳糖能够增加植物乳杆菌发酵乳中的活菌数、降低发酵乳的pH,并提高其持水力;响应面分析发现低聚半乳糖发酵乳的最佳制备条件为:2.0%的植物乳杆菌接种量、1.0%的低聚半乳糖添加量、发酵时间为24 h及发酵温度为42 ℃;添加低聚半乳糖的发酵乳能有效控制产肠毒素蜡样芽孢杆菌浓度在106 CFU/mL以下。该研究结果为低聚半乳糖及植物乳杆菌ZDY2013在发酵乳中的应用奠定了理论基础。     

A radiochemical assay for biotin in biological materials

A radiochemical assay for biotin is described. The assay is sensitive to one nanogram and simple enough for routine biotin analyses. The assay yields results which are comparable to those obtained from a microbiological assay using Lactobacillus plantarum.     

SPR技术检测奶粉中生物素

目的利用表面等离子共振(surface plasmon resonance,SPR)技术,建立快速定量测定牛奶中生物素的方法。方法将生物素共价偶联到表面等离子共振芯片CM5表面,并对竞争结合的生物素结合蛋白的结合浓度及芯片的再生条件进行优化,检测芯片的稳定性。在无抗生素牛奶中添加系列质量浓度的生物素,利用免疫竞争抑制原理构建标准曲线,并对市售10个奶粉样品进行检测。结果制备的芯片稳定,50个循环相对标准偏差(relative standarddeviation,RSD)小于10%。日间批内同一样品差异为8.75%,该方法的检测限为0.1μg/100g,回收率为80.4%~91.2%。10个牛奶产品中生物素含量全部在固定的允许范围内。所建立的方法可以在4 h内完成样品的前处理和检测。结论该方法是一种简便、快捷的定量检测方法。     

A RAPID METHOD FOR DETERMINING BIOTIN ACTIVITY IN RAW MATERIALS FOR FERMENTATION

The biotin activity of a raw material towards yeast may consist of the sum of the activities of biotin, desthiobiotin, oxybiotin, biocytin and possibly other factors. Some of these substances only stimulate certain strains of yeast; on the other hand, the strains of lactobacilli generally used for estimation of biotin activity utilize only D-biotin. It is thus desirable to have a method available in which a manufacturer's own strain of yeast is used as test organism for biotin activity. A simple method for biotin determination based on the cup plate assay is described. Biotin-free agar is inoculated with yeast and a series of standard solutions of D-biotin and a series of dilutions of the raw material under investigation are placed in holes cut in the agar. After incubation, the zones of growth are measured and from their diameters the biotin activity can be calculated.     

当量生物素控制对谷氨酸棒杆菌发酵产L-异亮氨酸的影响

以谷氨酸棒杆菌YILM 1504为出发菌,研究了生物素对L-异亮氨酸产量的影响。基于多梯度生物素对比实验及中后期生物素外源添加实验,确定了发酵液中最佳生物素浓度及补料工艺。结果显示:在低浓度生物素发酵液中,最适生物素浓度为45μg/L,此时产酸达到42.5g/L,糖酸转化率达到最高,为13.4%;在大于60μg/L高浓度生物素发酵液中,产酸最高为22g/L,表明高浓度生物素不利于产酸发酵。在5L发酵罐中,初始生物素浓度为30μg/L,18,32,42h分别添加10g/L玉米浆(15μg/L生物素),54h产酸量达到了44.5g/L,比初始生物素浓度为30μg/L,后期不补料产酸量提高了49.8%。     

TRYPSIN IMMOBILIZATION ON DERIVATIZED CELLULOSE BEADS BY BIOSPECIFIC AVIDIN‐BIOTIN INTERACTION AND CHARACTERIZATION OF THE IMMOBILIZED ACTIVITY1

Trypsin was immobilized on cellulose beads using the biospecific and high affinity avidin‐biotin interaction. Trypsin and cellulose beads were biotinylated with sulfosuccinimidyl‐6‐(biotinamido) hexanoate (NHS‐LC‐biotin). Avidin and biotinylated trypsin were sequentially adsorbed to the biotinylated cellulose beads. A similar procedure was carried out using controlled‐pore glass (CPG) beads. The properties of the two trypsin bioreactors were examined and compared. The substrate used for the assay of trypsin activity was p‐tosyl‐L‐arginine methyl ester and the extent of biotinylation of biotinylated trypsin and of immobilized biotin on cellulose beads and on CPG beads were determined using the 2‐[4′‐hydroxyazobenzene]benzoic acid dye‐binding method. Biotinylated trypsin in solution retained about 82% of the specific activity of native trypsin. Cellulose beads contained 0.184 μmol/mL (1.15 μmol/g) biotin and CPG beads, 0.329 μmol/mL (0.987 μmol/g). After regeneration, the biotin contents became slightly lower, namely, 0,159 μmol/mL for cellulose beads and 0.315 μmol/mL for CPG beads. The specific activities of trypsin immobilized on cellulose beads and CPG beads were 32 U/mL (202 U/g) and 43 U/mL (130 U/g), respectively. These studies indicate that cellulose beads can be biotinylated for use as bioselective support.     

Effects of biotin supplementation on peripartum performance and metabolites of Holstein cows

Fifty-two multiparous Holstein cows were randomly assigned to receive 0 or 20 mg of biotin/d starting at an average of 16 d prepartum and then switched to 0 or 30 mg of biotin/d from calving through 70 d postpartum to determine whether supplemental biotin would affect cow performance, hepatic lipidosis, and plasma metabolites. Mean concentration of biotin in plasma sampled weekly was greater in cows fed biotin (4.3 vs. 9.4 nmol/L). Postpartum dry matter intake as a percentage of body weight (3.9% vs. 4.0%), milk production (35.8 vs. 34.8 kg/d), and milk fat concentrations (3.59% vs. 3.69%) were similar between treatment groups. Milk from biotin-supplemented cows tended to have a greater concentration of protein (2.73% vs. 2.83%). Concentrations of plasma nonesterified fatty acids were lower at wk 2 (652 vs. 413 microEq/mL) and 4 (381 vs. 196 microEq/mL) postpartum in cows fed supplemental biotin. However, mean plasma concentrations of beta-hydroxybutyric acid were not affected by biotin supplementation. Mean concentration of plasma glucose was greater for lactating cows fed supplemental biotin (63.4 vs. 66.6 mg/dL). Biopsies of liver were taken at 2, 16, and 30 d postpartum. The triacylglycerol concentration in liver (wet basis) tended to decrease at a faster rate after d 2 postpartum with biotin supplementation compared with control cows. The potential mechanisms that link improved glucose status and decreased lipid mobilization in cows supplemented with biotin warrant further investigation.     

Sensitive Monoclonal Antibody-based Sandwich ELISA for the Detection of Porcine Skeletal Muscle in Meat and Feed Products

A monoclonal antibody‐based sandwich enzyme‐linked immunosorbent assay (ELISA) was developed for the sensitive detection of porcine skeletal muscle in raw and heat‐processed meat and feed products. Heat treatment of meat samples up to 132 °C for 2 h did not affect the assay performance. The assay uses a pair of monoclonal antibodies (MAbs 8F10 and 5H9) specific to skeletal muscle troponin I (TnI). MAb 8F10, reacting to mammalian TnI, is the capture antibody and the biotin‐conjugated MAb 5H9, specific to porcine TnI, the detection antibody. The sandwich ELISA is able to detect 0.05% (w/w) of laboratory‐adulterated pork in chicken, 0.1% (w/w) pork in beef mixtures, 0.05% (w/w) pork meal in soy‐based feed, and 1% commercial meat and bone meal (MBM), containing an unknown amount of pork, in soy‐based feed. This new assay provides a rapid and reliable means to detect the contamination of meat and feed products with trace amounts of porcine muscle tissue to ensure product quality and safety.     

二次接种叠加生物素的谷氨酸发酵工艺研究

由于受到发酵罐溶氧条件的限制,在高浓度生物素的谷氨酸发酵中往往出现产酸与糖酸转化率不协调的现象,针对这一现象,研究了二次接种叠加生物素的谷氨酸发酵工艺。在试验所用的发酵罐中,采用8.0μg/L生物素浓度的培养基作为发酵基础培养,经过一段时间发酵后,接入第二次种子液以及3.0μg/L(发酵液初始体积)的生物素量,通过适当的发酵控制,产酸水平达到139.6g/L,糖酸转化率高达62.80%,单罐谷氨酸产量比一次接种添加8.0μg/L生物素的发酵工艺提高了15.78%。     

氯化铵及生物素对苹果白兰地挥发性香气成分的影响

采用顶空固相微萃取(HS-SPME)-气相色谱-质谱联用(GC-MS)技术分析添加氯化铵及生物素与无添加氮源对照发酵苹果白兰地酒中的香气成分。结果表明,250 mg/L氯化铵与10 μg/L生物素的交互作用使苹果白兰地发酵中酯类物质的相对含量从40.95%提高至64.94%,异戊醇的相对含量从14.79%降低至3.69%;辛酸乙酯在主要酯类物质中相对含量最高且随着生物素添加量的增加从17.07%增加至22.43%,构成了苹果白兰地特有的香味。     

BIOTIN-ACTIVE COMPOUNDS,THEIR EXISTENCE IN NATURE AND THE BIOTIN REQUIREMENTS OF YEASTS

After a short account of the discovery of biotin and the progress of early biotin research, the natural occurrence of biotin with particular consideration to the raw materials used in the fermentation industry and its products is described. Of the many known biotin derivatives, those appearing in nature and those which can be converted to biotin-active (or inactive) compounds by simple procedures are reported. Ways to by-pass the need for biotin in microbes is discussed. The importance of biotin in yeast production, the biotin requirements of yeast, and the effect of culture conditions on these requirements are reported. The close relationship between the participation of biotinylenzymes and the biotin requirements is noted.