Transcriptome and Gene Ontology (GO) Enrichment Analysis Reveals Genes Involved in Biotin Metabolism That Affect l-Lysine Production in Corynebacterium glutamicum

Corynebacterium glutamicum is widely used for amino acid production. In the present study, 543 genes showed a significant change in their mRNA expression levels in l-lysine-producing C. glutamicum ATCC21300 than that in the wild-type C. glutamicum ATCC13032. Among these 543 differentially expressed genes (DEGs), 28 genes were up- or downregulated. In addition, 454 DEGs were functionally enriched and categorized based on BLAST sequence homologies and gene ontology (GO) annotations using the Blast2GO software. Interestingly, NCgl0071 (bioB, encoding biotin synthase) was expressed at levels ~20-fold higher in the l-lysine-producing ATCC21300 strain than that in the wild-type ATCC13032 strain. Five other genes involved in biotin metabolism or transport—NCgl2515 (bioA, encoding adenosylmethionine-8-amino-7-oxononanoate aminotransferase), NCgl2516 (bioD, encoding dithiobiotin synthetase), NCgl1883, NCgl1884, and NCgl1885—were also expressed at significantly higher levels in the l-lysine-producing ATCC21300 strain than that in the wild-type ATCC13032 strain, which we determined using both next-generation RNA sequencing and quantitative real-time PCR analysis. When we disrupted the bioB gene in C. glutamicum ATCC21300, l-lysine production decreased by approximately 76%, and the three genes involved in biotin transport (NCgl1883, NCgl1884, and NCgl1885) were significantly downregulated. These results will be helpful to improve our understanding of C. glutamicum for industrial amino acid production.     

不同修饰基团的副溶血性弧菌适配体在磁性纳米颗粒表面的固定效果和反应活性研究

目的 研究副溶血性弧菌适配体的不同修饰基团对磁性纳米颗粒修饰效果和与副溶血性弧菌结合活性的影响。方法 采用一步合成法合成氨基修饰的磁性纳米颗粒,将其分别与氨基、生物素和羧基修饰的副溶血性弧菌适配体结合,通过紫外吸收间接分析适配体的固定率,并利用外加磁场分离目标物,平板涂布确定磁分离效果。结果 生物素修饰的副溶血性弧菌适配体与磁性纳米颗粒有更高的结合效率,为62.16%。在菌液浓度较低时,不同基团修饰的适配体与磁性纳米颗粒结合后对副溶血性弧菌的分离效果影响较小,但当菌液浓度为105 CFU/mL时,不同基团修饰对副溶血性弧菌分离效果差距明显,氨基修饰的适配体显示了更为优异的反应活性,分离率为69.24%,分别较生物素和羧基修饰的适配体高21.04%和16.26%。结论 氨基修饰的副溶血性弧菌适配体在磁性纳米颗粒表面固定后显示了更为优越的反应活性,为副溶血性弧菌分离和检测的相关应用研究提供参考。     

一种中空金/银双金属纳米颗粒的制备及其SERS性能

以银纳米颗粒为牺牲模板,利用Ag和HAu Cl4之间的置换反应,结合柠檬酸钠同步还原的方法制备了一种中空金/银双金属纳米颗粒。通过对颗粒形貌及局域表面等离子体共振(LSPR)的分析,初步研究了此类金/银纳米颗粒的生长机理,并对影响反应的因素进行了探讨。结果表明,通过控制反应条件可以实现对LSPR的精密调控。该类金/银双金属纳米颗粒可用作为SERS基底,苯硫酚在其表面增强因子可达107,并具有良好的信号重现性。该基底用于atto610标记的生物素与亲和素的SERS检测,检测限可达80 pg/m L。     

高效液相色谱法测定发酵液中生物素含量

采用高效液相色谱法测定发酵液中生物素的含量.结果表明,样品浓度在5～150 μg·mL-1范围内线性关系良好,最低检测限为0.01 μg·mL-1,精密度实验的RSD为0.21%,稳定性实验的RSD为6.6%,发酵液在15 h内检测结果无明显差别,加样回收率在95%～102%之间.该方法简便、快捷、准确、重现性好,可用于发酵液中生物素的含量测定.     

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.     

Testing white line strength in the dairy cow

The tensile strength of 576 pieces of white line horn collected over 6 mo from 14 dairy cows restricted to parity 1 or 2 was tested. None of the cows had ever been lame. Seven cows were randomly assigned to receive 20 mg/d biotin supplementation, and 7 were not supplemented. Hoof horn samples were taken from zones 2 and 3 (the more proximal and distal sites of the abaxial white line) of the medial and lateral claws of both hind feet on d 1 and on 5 further occasions over 6 mo. The samples were analyzed at 100% water saturation. Hoof slivers were notched to ensure that tensile strength was measured specifically across the white line region. The tensile stress at failure was measured in MPa and was adjusted for the cross-sectional area of the notch site. Data were analyzed in a multilevel model, which accounted for the repeated measures within cows. All other variables were entered as fixed effects. In the final model, there was considerable variation in strength over time. Tensile strength was significantly higher in medial compared with lateral claws, and zone 2 was significantly stronger than zone 3. Where the white line was visibly damaged the tensile strength was low. Biotin supplementation did not affect the tensile strength of the white line. Results of this study indicate that damage to the white line impairs its tensile strength and that in horn with no visible abnormality the white line is weaker in the lateral hind claw than the medial and in zone 3 compared with zone 2. The biomechanical strength was lowest at zone 3 of the lateral hind claw, which is the most common site of white line disease lameness in cattle.     

以环糊精为核的星形七臂官能化聚己内酯的制备与表征及生物素修饰

以含有7个羟基的乙酰化β-环糊精为引发剂,开环聚合4-氨基甲酸苄酯-ε-己内酯(CABCL)单体,得到星形七臂官能化聚己内酯(CDSPCABCL)。并使用酸脱法与钯碳氢解法对CDSPCABCL的甲酸苄酯保护基脱保护,得到星形聚氨基己内酯(CDPACL)。同时,探索了投料比、聚合温度、聚合时间对CDSPCABCL聚合度的影响,以及酸脱法的酸倍数与反应时间、钯碳氢解法的反应时间对CDPACL断链率与脱保护率的影响。通过1 H NMR、GPC、FT-IR与TGA表征各步骤产物。最后通过CDPACL的氨基与生物素的羧基发生酯化反应,得到生物素化的星形聚己内酯,其在癌细胞靶向、生物探针等领域有潜在的应用前景。     

紫杉醇生物素标记衍生物的合成

以生物素、紫杉醇和6-氨基己酸为原料,经酰化、水解、酯化反应等步骤合成具有长臂连接的标题化合物。在三乙胺催化下,生物素与N-羟基琥珀酰亚胺反应生成N-羟基琥珀酰亚胺生物素酯。在氯化亚砜、甲醇反应体系下,由6-氨基己酸合成6-氨基己酸甲酯。在三乙胺作用下,N-羟基琥珀酰亚胺生物素酯和6-氨基己酸甲酯反应生成生物素-氨基己酸甲酯;生物素-氨基己酸甲酯在Li OH·H2O作用下,水解反应得到6-生物素氨基己酸。以DMF为溶剂,在N,N'-二环己基碳二亚胺(DCC)、4-二甲氨基吡啶(DMAP)作用下,6-生物素氨基己酸与紫杉醇反应,生成标题化合物。标题化合物和重要中间体化合物采用IR、1HNMR、MS进行了表征。     

基于国标法和试剂盒法对乳粉质控样品 生物素含量的测定

目的 用国标法和试剂盒法对乳粉质控样品的生物素含量进行测定, 评价两种方法对生物素检测结果差异性。方法 根据两种方法中植物乳杆菌(Lactobacillus plantarum)对游离生物素的特异性和灵敏性, 测定菌株的生长情况从而得到乳粉中生物素的含量。结果 国标法测定所得标准曲线范围为0~1 ng/100 g(R2=0.9993), 试剂盒法测定所得的标准曲线范围是0~0.72 μg/100 g(R2=0.9983), 线性关系良好; 两种方法测定生物素的含量差异不显著(P＞0.05), 结果均符合要求并在给定值的范围内。结论 两种方法对质控样品检测结果准确, 稳定性好, 适用于乳粉中生物素的检测。     

Pantothenic Acid and Biotin Retention in Cooked Legumes

Samples of legumes were analyzed raw for panfothenic acid and biotin. Pooled samples were prepared in one of three cooking times: 20, 90 or 150 min. For each cooking time legumes were presoaked with a short or long soak treatment. Average pantothenic acid retention in those cooked 20 min was 76% without pre-soak, 33% for short, and 44% for long soak. Biotin retention was 95% without pre-soak, 86% for short and 90% for long soak. In the 90 min group, retention differed between short soak and long soak for pantothenic acid (40% vs 58%), but not for biotin (98% vs 92%). No difference (P > 0.05) occurred in retention between short soak and long soak in the 150 min group for either nutrient. Biotin losses were minimal compared to pantothenic acid and methods of preparation could affect vitamin retention in legumes.