β-Galactosidase with transgalactosylation activity from Lactobacillus fermentum K4

The LacLM β-galactosidase of Lactobacillus fermentum K4 is encoded by 2 consecutive genes, lacL (large subunit) and lacM (small subunit), that share 17 overlapping nucleotides. Phylogenetic analysis revealed that this enzyme was closely related to other Lactobacillus β-galactosidases and provided significant insight into its common and distinct characteristics. We cloned both the lacL and lacM genes of L. fermentum K4 and heterologously expressed each in Escherichia coli, although the recombinant enzyme was only functional when both were expressed on the same plasmid. We evaluated the enzymatic properties of this species-specific LacLM β-galactosidase and discovered that it acts as both a hydrolase, bioconverting lactose into glucose and galactose, and a transgalactosylase, generating prebiotic galacto-oligosaccharides (GOS). The recombinant β-galactosidase showed a broad pH optimum and stability around neutral pH. The optimal temperature and Michaelis constant (K_m) for the substrates o-nitrophenyl-β-d-galactopyranoside and lactose were, respectively, 40°C and 45 to 50°C and 1.31 mM and 27 mM. The enzyme activity was stimulated by some cations such as Na⁺, K⁺, and Mg²⁺. In addition, activity was also enhanced by ethanol (15%, wt/vol). The transgalactosylation activity of L. fermentum K4 β-galactosidase effectively and rapidly generated GOS, up to 37% of the total sugars from the reaction. Collectively, our results suggested that the β-galactosidase from L. fermentum K4 could be exploited for the formation of GOS.     

壳聚糖固定化绿豆乳糖酶的特性及应用的初步研究

以壳聚糖凝胶颗粒为载体,采用共价键偶联法固定绿豆乳糖酶,研究壳聚糖固定化绿豆乳糖酶的酶学性质并对其进行水解牛乳中乳糖的初步应用研究。结果表明：固定化酶的热稳定性在4～55℃范围内;最适温度60℃;最适pH3.5;固定化酶的表观米氏常数Km 为0.04%,溶液酶米氏常数Km 为0.33%,是溶液酶米氏常数Km 的0.12 倍;固定化酶的操作半衰期为30d,较溶液酶长。用固定化酶制备填充床式反应器水解牛奶中乳糖,水解率可达60% 以上。     

Effect of lactose content on dielectric properties of whole milk and skim milk

Dielectric properties (dielectric constant ε′ and dielectric loss factor ε′′) of whole milk and skim milk with the lactose content of 4.56–6.44% and 4.57–6.47%, respectively, were measured over the frequency range of 20–4500 MHz at 25 °C using a vector network analyzer. The results showed that ε′ decreased with increasing frequency, and ε′′ changed with V shape and its minimum was noted at about 2000 MHz. Whole milk had lower dielectric properties than skim milk at almost same lactose content and a given frequency. ε′ had weak positive linear relationship with lactose content for whole milk, but had negative linear relationship for skim milk. No matter for which milk, ε′′ had very good negative linear relationship with lactose content below 1000 MHz and had good positive linear relationship above 2300 MHz. The study provides information on developing rapid lactose detector for milk in future.     

Yeast on the milky way: genetics,physiology and biotechnology of Kluyveromyces lactis

The milk yeast Kluyveromyces lactis has a life cycle similar to that of Saccharomyces cerevisiae and can be employed as a model eukaryote using classical genetics, such as the combination of desired traits, by crossing and tetrad analysis. Likewise, a growing set of vectors, marker cassettes and tags for fluorescence microscopy are available for manipulation by genetic engineering and investigating its basic cell biology. We here summarize these applications, as well as the current knowledge regarding its central metabolism, glucose and extracellular stress signalling pathways. A short overview on the biotechnological potential of K. lactis concludes this review. Copyright © 2013 John Wiley & Sons, Ltd.     

Detection of glucose,galactose, and lactose in milk with a microdialysis-coupled flow injection amperometric sensor

A microdialysis-coupled flow injection amperometric Sensor (microFIAS) was used to determine glucose, galactose, and lactose in milk. The sensor is based on enzyme-catalyzed reaction in combination with the three well-established analytical techniques, namely; microdialysis sampling, flow injection analysis (FIA), and amperometric detection. With the multianalyte sensor it was possible to detect glucose and galactose by sequential injection of their corresponding oxidase enzymes: glucose oxidase and galactose oxidase, while lactose was determined by injection of a mixture of beta-galactosidase and glucose oxidase enzymes. The sensor showed a linear response between 0.05 and 10 mM for glucose, between 0.1 and 20 mM for galactose and between 0.2 and 20 mM for lactose, respectively. The relative standard deviation values of the sensor measurements for glucose, galactose, and lactose were 3-4% (n = 3). The sensor measurements for lactose content in milk were compared with a standard method with an infrared spectrophotometer.     

Influence of manufacturing process on tabletting ability of powder: comparison between blending, grinding, and spray drying of two formulations made of theophylline and lactose/cellulose or Cellactose

Three manufacturing processes were applied to two formulations composed of 20% anhydrous theophylline associated with either 20% microcrystalline cellulose and 60% lactose or 80% Cellactose®. The processing method (dry blending, grinding, or spray drying) and the formulation were investigated through the comparison of the physical and flow characteristics and the compactibility of the end products. The results demonstrated that the formulation had a major effect on the mechanical properties, with binary blends exhibiting a higher resistance than ternary ones, whereas flow properties and densification depended on the process. Nevertheless, it was also observed that spray drying decreased the difference between the mechanical properties of the two formulations, probably by modifying the texture of the Cellactose® in suspension.     

光折射法对牛乳乳糖的快速精确测量

针对我国乳制品中乳糖质量含量的测量准确度偏低，操作复杂的情况，依据光折射理论，提出了牛乳乳糖质量含量的快速高准确度测量方法，实际测量和定标实验表明，本方法测量牛乳乳糖的质量分数可达到0.1%的准确度。     

酵母乳糖酶对牛乳乳糖水解作用的研究

采用乳酸克鲁维酵母的乳糖酶(β-乳糖苷酶)对乳糖溶液、喷雾干燥脱脂奶以及全脂奶进行了试验,以确定将乳糖转化成单糖的最适条件。当牛奶中酶浓度为3u/ml时,于40℃反应2小时或4℃反应24小时,约60％的乳糖得以水解。随着乳糖浓度的增高,水解程度也有所提高。当酶浓度为10U/ml时,于40℃反应2小时,90％乳糖得以水解。用自制的乳酸克鲁维酵母的乳糖酶和加拿大lactaid Inc.生产的乳糖酶制剂lactaid水解乳糖无大的差异。     

分光光度法测定发样中的微量铁

建立了以邻二氮菲测定人发样中的微量铁的分光光度法。研究表明,在pH=2～9的溶液中,铁（Ⅲ）可用盐酸羟胺还原至铁（Ⅱ）,生成的铁（Ⅱ）可与邻二氮菲生成稳定的橘红色络合物,其最大吸收波长为506 nm,铁浓度在0.31～16.00 mg/L范围内与溶液吸光度呈良好线性关系,线性回归方程为A=0.20192ρ-0.01164（mg/L）,相关系数r=0.9999,检出限为0.157 mg/L,相对标准偏差（RSD）为0.58%,表观摩尔吸收系数ε=3.59×104L.（mol-1cm-1）。     

Production, health aspects and potential food uses of dairy prebiotic galactooligosaccharides

Galactooligosaccharides are sugars composed of 3-10 molecules of galactose and glucose via a transgalactosylation reaction mediated by the enzyme β-galactosidase. Prebiotics are non-digestible food ingredients that pass through the upper digestive system relatively intact and ferment in the lower colon, producing short-chain fatty acids that support the growth of supplemented or indigenous colonic microbiota. Galactooligosaccharides and other prebiotic ingredients are increasingly being recognized as useful dietary tools for the modulation of the colonic microflora toward a healthy balance. Galactooligosaccharides compare well to other oligosaccharides in terms of their prebiotic, immunomodulation, and functional properties in foods. This review elucidates the galactooligosaccharide production process from refined lactose and/or cheese whey permeates, galactooligosaccharide market share and economic value, their health properties, and potential food applications.