Design of Trehalose-Based Amide/Sulfonamide C-type Lectin Receptor Signaling Compounds

Mincle agonists have been shown to induce inflammatory cytokine production, such as tumor necrosis factor-alpha (TNF) and promote the development of a Th1/Th17 immune response that might be crucial to development of effective vaccination against pathogens such as Mycobacterium tuberculosis. As an expansion of our previous work, a library of 6,6′-amide and sulfonamide α,α-d -trehalose compounds with various substituents on the aromatic ring was synthesized efficiently in good to excellent yields. These compounds were evaluated for their ability to activate the human C-type lectin receptor Mincle by the induction of cytokines from human peripheral blood mononuclear cells. A preliminary structure–activity relationship (SAR) of these novel trehalose diamides and sulfonamides revealed that aryl amide-linked trehalose compounds demonstrated improved activity and relatively high potency cytokine production compared to the Mincle ligand trehalose dibehenate adjuvant (TDB) and the natural ligand trehalose dimycolate (TDM) inducing dose-dependent and human-Mincle-specific stimulation in a HEK reporter cell line.     

透性化酒精酵母细胞产海藻糖发酵培养基的优化

为提高酒精酵母细胞发酵液中海藻糖的含量,在制备出透性化酒精酵母细胞的基础上,用单因素实验和响应面分析法,优化了酒精酵母的发酵培养基,确定了最适培养基组成.实验结果表明,酵母膏、蔗糖及氯化钠的添加量对海藻糖积累影响显著,影响程度依次为:酵母膏＞蔗糖＞氯化钠.优化后的培养基组成为:酵母膏14.7 g/L,蔗糖32.5 g/...     

高效液相色谱法测定海藻糖

采用高效液相色谱法（HPLC），以Hypersil NH2柱为分析柱，乙腈-水（4：1）为流动相，采用示差折光检测器，一次进样能同时分离样品中的果糖，葡萄糖，蔗糖，麦芽糖，海藻糖，低聚糖等。测定了样品中的葡萄糖，麦芽糖，海藻糖的含量，线性相关系数0.9998-0.9999，变异系数0.67%-1.0%。回收率98.65%-100.6%。     

外源相容性溶质对S酵母耐盐性及酱油风味形成的研究

研究外源相容性溶质对S酵母(Zygosaccharomyces rouxii)耐盐性的影响,并探讨在高盐稀态酿造工艺中添加海藻糖对酱油风味的影响.结果表明,在酵母培养基中添加0.005mol/L海藻糖、甜菜碱和山梨醇,均可以显著改善S酵母在高盐培养基中的生长情况,缩短渗透胁迫条件下S酵母的延滞期,增加生物量,其中海藻糖的渗透保护效果最佳;添加海藻糖可使酱油的主体香味物质HEMF(4-羟基-2-乙基-5-甲基-3-呋喃酮)和乙酸乙酯的含量显著增加,同时促进S酵母合成酱油中典型风味物质4-EG (4-乙基愈创木酚).在酱油高盐稀态酿造中添加相容性溶质,如海藻糖,可有效改善酱油的风味和品质.     

海藻糖、乳酸钠对冻藏鳙鱼鱼糜蛋白抗冻效果的影响

以凝胶强度、盐溶性蛋白含量、Ca~(2+)-ATPase活性、总巯基、表面疏水性为指标并结合DSC分析(差示扫描量热分析),研究了海藻糖、乳酸钠在8%(w/w)水平上对鳙鱼鱼糜蛋白在-18℃冻藏24周的抗冻效果,并将其抗冻效果与传统商业抗冻剂(4%蔗糖+4%山梨糖醇)进行了对比。结果表明,海藻糖、乳酸钠的加入能较好地抑制冻藏鳙鱼鱼糜蛋白的冷冻变性,表现在这2种物质加入后,冻藏鳙鱼鱼糜的凝胶能力得到较好的维持,盐溶性蛋白含量、Ca~(2+)-ATPase活性和巯基含量的下降及表面疏水性的增加也得到很好的抑制,2种物质的抗冻效果比传统商业抗冻剂具有明显的优势。DSC分析表明,抗冻剂的添加可能改变了鳙鱼鱼糜肌球蛋白的结构,使其变得更稳定,从而表现出较好的冻藏稳定性。海藻糖、乳酸钠都属于低甜度、低热值物质,因此在鱼糜加工业中有望成为传统商业抗冻剂的替代物。     

海藻糖合成酶产生菌筛选、鉴定及其产酶特性

以麦芽糖为唯一碳源高盐培养基,经高温培养,从温泉水样及其附近土壤中筛选得到一株菌株T2,经过生物合成途径初步验证,该菌株产海藻糖合酶,能够通过海藻糖合成酶（TreS）途径将麦芽糖转化为海藻糖。菌株T2为革兰氏阳性菌,杆状,有芽孢,经过生物学鉴定,将其初步鉴定为芽孢杆菌属（Bacillus sp.）。对其产海藻糖合酶的酶学性质进行了研究：酶反应最适作用温度为60 ℃,在60 ℃条件下保温100 min仍能保持酶活性80.7%;最适作用pH值为7.0,在pH 6.0～7.5范围内稳定。采用正交试验对其发酵培养基配方进行了优化研究,确定了最佳的培养基组成为牛肉膏3.0 g/L,麦芽糖20.0 g/L,蛋白胨7.5 g/L,无机盐（K2HPO4＋NaH2PO4＋MgSO4·7H2O）3.0 g/L。在此条件下,菌株T2产海藻糖合酶酶活力达到310.6 U/L。     

Preservation of biological materials under desiccation

A number of life forms, including seeds, certain nematodes, bacterial and fungal spores, and cysts of certain crustaceans, show an ability to survive desiccation. The present article reviews the literature available on this subject and critically evaluates the evidence for various mechanisms that may be responsible for these phenomena. Specific mechanisms considered include vitrification (glass formation) by sugars and other polyhydroxy compounds that are accumulated by the desiccated structures, specific effects of polyhydroxy compounds on membranes, effect of “compatible solutes” on conformation of key proteins, as well as other biochemical mechanisms.

The article presents potential applications relevant to food technology and to biotechnology and reviews the research required to materialize more effective use of desiccation in food and biopreservation.     

HPLC在研究微杆菌D-97胞内酶合成海藻糖机制中的应用

微杆菌D 97胞内酶对麦芽糖的水解微弱 ,并且合成海藻糖的酶反应对磷酸盐无依赖性。采用氨基键合柱HPLC对酶作用的初始底物麦芽寡糖混合物 (Ⅰ )、酶作用 2 4h后未糖化的底物 (Ⅱ )及其糖化水解后的底物 (Ⅲ )分别进行检测对照 ,底物Ⅰ麦芽五糖占34 1 5 % ,底物Ⅱ海藻糖、三糖和五糖分别占 1 7 68%、37 63 %和 3 0 8% ,底物Ⅲ的 2种主要产物葡萄糖和海藻糖各占 77 57%、2 0 2 0 %。通过上述 3种底物二糖至六糖浓度变化的比较 ,发现 DP 值 >4的麦芽寡糖为微杆菌D - 97胞内酶合成海藻糖的较适底物 ,并推断麦芽五糖生成海藻糖的基本方式为 :麦芽五糖→麦芽三糖 +海藻糖。以麦芽五糖为底物的酶反应研究进一步证实了上述作用方式。对于筛选得到的野生菌酶 ,HPLC对照检测从 1个角度为其酶合成海藻糖基本机制研究提供 1种方法。     

海藻糖和甘露醇对冻融循环引起的虾蛄肌原纤维蛋白结构和功能特性变化的影响

研究4 g/100 mL海藻糖＋4 g/100 mL甘露醇对反复冻融引起的虾蛄肌原纤维蛋白结构和功能特性变化的影响,为海产品保鲜提供理论依据。本实验对添加保护剂的实验组和未添加保护剂的对照组分别进行0、1、3、5 次冻融处理,通过测定蛋白羰基、总巯基、表面疏水性、内源色氨酸荧光强度、弹性储能模量G′、凝胶强度和持水力、凝胶微观结构、乳化活性和乳化稳定性等,探究海藻糖和甘露醇的抗冷冻变性作用。结果显示,随着冻融次数的增加,蛋白羰基含量和表面疏水性显著增加（P＜0.05）,总巯基含量和色氨酸荧光强度显著下降（P＜0.05）,蛋白出现不同程度聚集,凝胶和乳化性能降低。海藻糖和甘露醇的添加有效抑制蛋白的变性程度,减少蛋白结构的变化,显著提高蛋白在热凝胶过程中的G′,改善肌原纤维蛋白的凝胶特性（凝胶强度、持水力和三维网络结构）、乳化活性和乳化稳定性。海藻糖和甘露醇能与蛋白质功能基团结合,使蛋白质分子处于饱和状态从而避免聚集;另外通过束缚水分子,降低“共晶点”温度,减少冰晶体的形成量,隔离和减缓蛋白质聚集,防止蛋白质凝聚变性,从而改善冻藏过程中的虾蛄肌肉品质。     

Supplementation of the diet of dairy cows with trehalose results in milk with low lipid peroxide and high antioxidant content

The objective of this study was to investigate the effect of dietary supplementation with the disaccharides trehalose and cellobiose on antioxidant activity in rumen fluid, blood, and milk of dairy cows. Nine Holstein dairy cows housed in a free-stall barn were divided into 3 groups, with each group receiving a different dietary treatment (a control diet, a 1% trehalose-supplemented diet, or a 1% cellobiose-supplemented diet) following a 3 × 3 Latin square design. Feed intake and milk production increased in cows receiving the trehalose-supplemented diet compared with those receiving the control and cellobiose-supplemented diets. The total protozoa numbers in the rumen fluid of cows fed trehalose- or cellobiose-supplemented diets were greater than those of the control group. The C18:0 and C18:1 fatty acid content was increased in the milk of cows fed the trehalose-supplemented diet compared with that of the control group, and the C18:3n-3 fatty acid content in the milk of cows fed the cellobiose-supplemented diet was less than that of the control group. Plasma biochemical parameters were unchanged among the different treatments. In rumen fluid, 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and superoxide dismutase activity were increased 2 h after feeding in cows receiving the cellobiose-supplemented diet compared with the control group, and the concentration of thiobarbituric acid reactive substances in the rumen fluid of cows fed the cellobiose-supplemented diet was decreased. In contrast, the values of these parameters measured in the milk of cows fed the cellobiose-supplemented diet were no different from those of control cows. Dietary supplementation with trehalose did, however, bring about an improvement of the oxidative status of milk and blood in these animals compared with controls. These results provide the first evidence supporting the use of dietary disaccharides to decrease lipid peroxide levels and increase the antioxidant content of dairy cow milk. The findings suggest that disaccharides, particularly trehalose, might be useful as supplements for reducing oxidative stress and improving the quality of milk for human consumption, as well as possibly impairing the processes that give rise to lipid oxidation odor in dairy cow milk.