超滤膜分离纯化加纳籽中5-羟基色氨酸的研究

研究了超滤膜从加纳籽中分离纯化5-羟基色氨酸的相关膜工艺条件及参数,并筛选出适合的聚砜超滤膜。研究结果表明,截留分子量为50 K的超滤膜分离纯化5-羟基色氨酸的效果要优于截留分子量分别为10 K和100 K的超滤膜。超滤膜分离纯化5-羟基色氨酸优化工艺条件:操作压力0.10 MPa、操作温度35℃、料液质量浓度1.141 mg/m L、p H值7.0,在此条件下5-羟基色氨酸转移率为83.5%,纯度可达90.5%。     

超滤膜分离纯化加纳籽中5-羟基色氨酸的研究北大核心CSCD

研究了超滤膜从加纳籽中分离纯化5-羟基色氨酸的相关膜工艺条件及参数,并筛选出适合的聚砜超滤膜。研究结果表明,截留分子量为50 K的超滤膜分离纯化5-羟基色氨酸的效果要优于截留分子量分别为10 K和100 K的超滤膜。超滤膜分离纯化5-羟基色氨酸优化工艺条件:操作压力0.10 MPa、操作温度35℃、料液质量浓度1.141 mg/m L、p H值7.0,在此条件下5-羟基色氨酸转移率为83.5%,纯度可达90.5%。     

美洲大蠊愈创活性成分研究北大核心CSCD

采用色谱分离手段从美洲大蠊中分离得到17个化合物,利用波谱解析鉴定了它们的结构,分别命名为环(酪氨酸-酪氨酸)(1)、环(酪氨酸-脯氨酸)(2)、环(缬氨酸-酪氨酸)(3)、环(甘氨酸-苯丙氨酸)(4)、环(甘氨酸-色氨酸)(5)、环(色氨酸-丝氨酸)(6)、环(色氨酸-天冬酰胺)(7)、ginsenine(8)、6-羟基香豆素(9)、6-羟基色满-2-酮(10)、1-(3-乙基苯基)-1,2-乙二醇(11)、1-(4-乙基苯基)-1,2-乙二醇(12)、(E)-3,4-二羟基苯亚甲基丙酮(13)、2-羟基-3',4'-二羟基苯乙酮(14)、原儿茶酸(15)、对羟基苯甲酸甲酯(16)和丁香酸(17)。其中,化合物1~14均为首次从美洲大蠊中分离得到。此外,对化合物促进创面愈合作用进行了观察,化合物13显示较强的抑制NO生成活性。     

美洲大蠊愈创活性成分研究

采用色谱分离手段从美洲大蠊中分离得到17个化合物,利用波谱解析鉴定了它们的结构,分别命名为环(酪氨酸-酪氨酸)(1)、环(酪氨酸-脯氨酸)(2)、环(缬氨酸-酪氨酸)(3)、环(甘氨酸-苯丙氨酸)(4)、环(甘氨酸-色氨酸)(5)、环(色氨酸-丝氨酸)(6)、环(色氨酸-天冬酰胺)(7)、ginsenine(8)、6-羟基香豆素(9)、6-羟基色满-2-酮(10)、1-(3-乙基苯基)-1,2-乙二醇(11)、1-(4-乙基苯基)-1,2-乙二醇(12)、(E)-3,4-二羟基苯亚甲基丙酮(13)、2-羟基-3',4'-二羟基苯乙酮(14)、原儿茶酸(15)、对羟基苯甲酸甲酯(16)和丁香酸(17)。其中,化合物1~14均为首次从美洲大蠊中分离得到。此外,对化合物促进创面愈合作用进行了观察,化合物13显示较强的抑制NO生成活性。     

氨基酸类及其衍生物类的酶抑制剂

<正> 从细菌、动物、植物体中分离得到240多种氨基酸,其中只有20种用于蛋白质合成,另外一些象乌氨酸、瓜氨酸、5羟基色氨酸、二羟基苯丙氨酸和高丝氨酸是代谢的中间产物,生物有自生的保护作用,防止其他生物的侵害,其主要机制是氨基酸及其衍生物产生了对酶的抑制。     

DL-5-甲基色氨酸的化学合成

<正> DL-5-甲基色氨酸是色氨酸的一种衍生物,它和其它衍生物一起(如DL-5-氟-色氨酸)主要用于处理色氨酸发酵产生菌,从而获得抗类似物高产菌株,在此基础上,利用重组DNA技术,进一步获得“工程菌株”,因此,它是色氨酸发酵科研中的一     

岩木瓜茎干化学成分及PTP1B抑制活性研究

综合运用天然产物化学分离、纯化技术从岩木瓜茎干中分离得到18个化合物,结合各化合物理化性质和光谱数据鉴定其结构,依次为对羟基苯乙酮(1)、对羟基苯甲酸(2)、对羟基苯甲酸乙酯(3)、3-hydroxy-1-(4-hydroxy-3-methoxyphenyl)propan-1-one(4)、咖啡酸(5)、咖啡酸乙酯(6)、对羟基苯甲醛(7)、咖啡酸甲酯(8)、对羟基桂皮酸(9)、水杨酸(10)、原儿茶酸(11)、3,5-二羟基-4-甲氧基苯甲酸(12)、对醛基苯基-1-O-葡萄糖苷(13)、色氨酸(14)、豆甾-3,6-二酮(15)、β-谷甾酮(16)、5α-9(11)-豆甾烯-3β-醇(17)和胡萝卜苷(18)。这些化合物均为首次从该植物分离得到,其中化合物4,6,16和17为首次从该属植物中分离得到。对从岩木瓜中分离得到的46个化合物进行PTP1B抑制活性筛选,11个化合物具有PTP1B抑制活性,IC50值在1.0~37.0μM之间。     

维药瘤果黑种草籽化学成分研究

为研究维药瘤果黑种草籽的化学成分,采用硅胶、ODS柱色谱等方法分离纯化,分离并鉴定了16个化合物,利用理化性质及波谱方法鉴定化学成分的结构分别为:Nigeglanine(1)、附子碱(2)、L-色氨酸(3)、L-色氨酸甲酯(4)、5,7-二羟基-异苯并呋喃酮(5)、豆甾醇(6)、豆甾醇-3-O-β-D-葡萄糖苷(7)、β-谷甾醇(8)、β-胡萝卜苷(9)、亚油酸(10)、邻苯二甲酸二丁酯(11)、棕榈酸(12)、十六烷酸1-甘油酯(13)、硬酯酸(14)、硬酯酸2-甘油酯(15)和蔗糖(16)。其中化合物3～5、7、9、11和15为首次从该种药材中得到。     

色氨酸, 天然产物生物合成中的重要结构单元

氨基酸作为生物体内组成生命物质的小分子化合物, 在天然产物生物合成中扮演了非常重要的作用。色氨酸含有一个独特的吲哚环, 相对复杂的吲哚环平面结构使得色氨酸相比其他氨基酸具有更多的修饰空间。在微生物天然产物生物合成研究中, 色氨酸及其衍生物经常作为组成模块参与到天然产物的生物合成中, 本文概述了色氨酸几种不同的生物修饰方式, 包括烷基化修饰、卤化修饰、羟基化修饰、以及吲哚环的开环重排反应等。分析并总结色氨酸在天然产物生物合成中的作用可以增加我们对天然产物结构多样性的认识和推动天然产物生物合成机制的研究。     

灰毛豆叶片的化学成分研究

从灰毛豆叶片中分离得到10个化合物,通过波谱数据和理化性质分别鉴定为:stigmast-1,5-dien-3β-ol(1),clemaphenol A(2),松脂醇(3),桉素(4),3-吲哚甲醛(5),色氨酸(6),(S)-4-苄基-2-噁唑烷酮(7),4-羟基苯基丙酸(8),buteaspermanol(9),bovatachalcone(10)。其中化合物1、2、4、5、7、8、9首次从该植物中分离得到。     

超滤法和盐析法粗提基因工程干扰素其比活的比较

我们的工作是应用ＹＦ２ＰＳＫ３１Ｒ型超滤器超滤粗提基因工程干扰素,并与盐析法对比。超滤法简便快速条件温和,超滤后干扰素平均收率为９３．９５％±２．４８％,盐析法则为７５．９３％±３．９％,前者比后者提高１８．０２％,差异显著。比活前为：４．４５５Ｅ６±７．５８Ｅ５ＩＵ／ｍｇ蛋白,后者为：１．６１１７Ｅ６±２．０７９Ｅ５ＩＵ／ｍｇ蛋白,差异也是显著的     

Stereoselective HPLC separation of alvimopan on cellulose‐based immobilized polysaccharide as a chiral stationary phase

Chiral separation by normal phase high performance liquid chromatography is one of the most powerful technique to quantify the chiral purity of the compounds. In this study, a novel, simple, and specific analytical method was proposed to ascertain the chiral purity of alvimopan (ALV). The normal phase HPLC method was developed based on cellulose tris (3,5‐dichlorophenylcarbamate) stationary phase. The separation of ALV isomers achieved by using column CHIRALPAK IC (250 × 4.6 mm, 5 μm), mobile phase n‐hexane: isopropyl alcohol: ethanol: diethylamine (650:200:150:5 v/v), column oven temperature 30°C, flow rate 1.0 mL min^?1, injection volume was 10 μL, chromatographic response monitored at 273 nm. The developed method was validated as per the ICH guidelines and found precise, accurate, and linear. The advantage of the method is a good separation of ALV isomers within 35 minutes of the analysis time. Therefore, this method is suitable for routine determination of chiral purity of ALV active pharmaceutical ingredient.     

Two methods for determination of transketolase activity

Two new optical methods for transketolase activity assay using only one substrate, xylulose 5-phosphate or glycol aldehyde, have been developed. For transketolase activity assay in the first method, it is necessary to add auxiliary enzyme, glyceraldehyde phosphate dehydrogenase. It is not needed in the second method. The range of transketolase concentration in the activity assay is 0.036–0.144 U/ml for the first method and 1.8–6.8 U/ml for the second one. Published in Russian in Biokhimiya, 2006, Vol. 71, No. 5, pp. 693–696.     

两种精原干细胞高效能纯化方法的比较研究*

目的: 比较贴壁分离法和免疫磁珠法纯化小鼠精原干细胞(mSSCs) 的优缺点。方法: 分别选取10只12-15日龄的雄性C57BL/6小鼠,颈椎脱臼法处死,摘取睾丸用酶消化法获得曲细精管单细胞悬液,分别用贴壁分离法和免疫磁珠法从单细胞悬液中分离纯化mSSCs,并针对两种方法在细胞数量、分离效率以及对细胞增殖生长的影响等方面进行比较。结果: 两种纯化方法均可从小鼠曲细精管单细胞悬液中分离纯化得到干细胞,并可在体外培养后呈现出典型的精原干细胞特有的葡萄串状克隆,体外连续培养增殖超3个月。10只小鼠的睾丸经差异贴壁法纯化后可以得到3×10⁵±0.4×10⁵个mSSCs(n=5),细胞回收率(纯化后细胞数/曲细精管单细胞悬液细胞数)为1.5%±0.1%(n=5);经免疫磁珠法可以得到6×10⁵±0.4×10⁵个mSSCs(n=5),细胞回收率为3.0%±0.1%(n=5),免疫磁珠法得到的干细胞数量更高。差异贴壁法得到的干细胞更纯,因为体外培养5 d左右即得到干细胞集落,而免疫磁珠法得到的干细胞则约10 d才可以看到明显的细胞集落,但是两种纯化方法对细胞体外长期增殖生长没有明显的影响。结论: 两种方法均可以纯化得到高质量的mSSCs,,但两种方法各有优缺点。差异贴壁法较免疫磁珠法经济、实用,无需购买专门的设备和抗体磁珠,但获得的细胞数量相对较低,用时也较长。     

Silicon: An indigestible marker for measuring food consumption and utilization by insects

Silicon (Si) is a useful indigestible marker in plants for measuring food consumption and nutritional indices. Si concentrations in HF:HCl extracts of small (5–20 mg) samples may be measured easily using atomic emission spectroscopy. The Si marker and gravimetric methods were compared usingLymantria dispar (L.) larvae fed artificial diet with 2.5% silica (SiO₂). Both methods produced equivalent measures of consumption and nutritional indices. The Si marker method was also used to measure the consumption of six grass species by larvae ofParatrytone melane Edwards. The results were similar overall when compared with estimates of consumption based on total fecal weight, but varied significantly for some grass species. The Si marker method may be a useful alternative to the gravimetric method, especially in field situations.     

一种简便制作小型鱼类形态标本的方法

本文介绍一种简便制作小型鱼类形态标本的方法.包括以下步骤:1.制作前准备;2.剥离皮肤;3.支撑架制作与放置;4.缝合;5.配制填充材料;6.填充;7.装义眼与整理姿态;8.干燥,上色,生境制作与固定.与其他制作小型鱼类形态标本的方法相比,新方法中先缝合再填充的步骤,难度小,容易控制填充物的量,做出的标本形态更逼真.     

注射用转移因子不同生产方法比较

比较从猪脾中提取活性转移因子的不同生产方法,以建立适合产业化的制造工艺。将猪脾匀浆后,分别冻融0、3、5次,再以超滤法或Lawence法提取转移因子。与lawrence法相比,经5次冻融并超滤制备的产品收率高,符合国家标准。匀浆冻融次数对转移因子的收率和质量有一定的影响,超滤法适合于转移因子的大规模生产。     

PCR-DGGE detection of the bacterial community structure in the Inner Mongolia steppe with two different DNA extraction methods

Compared with conventional methods, molecular biological technique, such as PCR-DGGE (denaturing gradient gel electrophoresis), is informative in examining the structure of the soil bacterial community through the extraction of microbial DNA from soil and generation of bacterial community profiles by PCR amplification of 16S rRNA genes. Extraction efficiency of soil microbial DNA is the most important step in these methods. At present, the frozen-thawing method and bead-beating method are most widely used in genomic extraction. Nevertheless, comparison of these two methods has not been conducted in different soil types, especially in humus-rich soil. In this study, extraction efficiencies of the two methods were compared in humus-rich steppe soil in Inner Mongolia based on the PCR-DGGE analysis of bacterial community structure. The results indicated that the bead-beating method is better than the frozen-thawing method in genomic DNA extraction efficiency. In addition, 21 bands in the DGGE pattern with the bead-beating method were further selected, cloned and sequenced. Based on similarity matching, all the sequences formed five major clusters: Actinobacteria; α-, β-, γ-, Proteobacteria ; Bacteriodetes ; Gemmatimonadetes and Acidobacteria . Of the 21 clones obtained from DGGE patterns, YC4 showed 99.7% similarity to Pseudomonas sp. (DQ339153); YC5, YC18 and YC19 showed 99.9 % similarity to Gram-positive bacterium (AB008510), Virgisporangium ochraceum (AB006162) and Micromonospora chalcea (X92613), respectively.     

Detection of Salmonella enterica in food using two-step enrichment and real-time polymerase chain reaction

Aims: A new real‐time polymerase chain reaction‐based method was developed for the detection of Salmonella enterica in food. Methods and Results: The method consisted of a novel two‐step enrichment involving overnight incubation in buffered peptone water and a 5‐h subculture in Rappaport–Vassiliadis medium, lysis of bacterial cells and a Salmonella‐specific 5′‐nuclease real‐time PCR with an exogenous internal amplification control. Because a two‐step enrichment was used, the detection limit for dead S. enterica cells in artificially contaminated ice cream and salami samples was high at 10⁷ CFU (25 g)^?1, eliminating potential false‐positive results. When the method was evaluated with a range of 100 naturally contaminated food samples, three positive samples were detected by both the real‐time PCR‐based method and by the standard microbiological method, according to EN ISO 6579. When the real‐time PCR‐based method was evaluated alongside the standard microbiological method according to EN ISO 6579 with 36 food samples artificially contaminated at a level of 10⁰ CFU (25 g)^?1, identical results were obtained from both methods. Conclusions: The real‐time PCR‐based method involving a two‐step enrichment produced equivalent results to EN ISO 6579 on the day after sample receipt. Significance and Impact of the Study: The developed method is suitable for rapid detection of S. enterica in food.     

Quantitation of heparosan with heparin lyase III and spectrophotometry

Heparosan is Escherichia coli K5 capsule polysaccharide, which is the key precursor for preparing bioengineered heparin. A rapid and effective quantitative method for detecting heparosan is important in the large-scale production of heparosan. Heparin lyase III (Hep III) effectively catalyzes the heparosan depolymerization, forming unsaturated disaccharides that are measurable using a spectrophotometer at 232 nm. We report a new method for the quantitative detection of heparosan with heparin lyase III and spectrophotometry that is safer and more specific than the traditional carbazole assay. In an optimized detection system, heparosan at a minimum concentration of 0.60 g/L in fermentation broth can be detected.