羧甲基茯苓多糖的水媒法制备及其免疫活性研究

羧甲基茯苓多糖carboxymethyl-pachymaran（CMP）具有良好的免疫调节活性,传统方法使用两步工序在醇相体系中制得,有机溶剂消耗大、成本高。本研究报道一种新制备方法,将提取茯苓多糖以及茯苓多糖与氯乙酸的醚化反应整合在一个碱液体系中完成。动物实验表明,新方法制备的CMP能提高环磷酰胺诱导的免疫低下小鼠的细胞免疫和体液免疫功能,对S180肉瘤具有抑制活性。经口急性毒理结果显示CMP属于实际无毒物质,适合于功能性食品和保健食品开发应用。     

液体发酵茯苓胞外多糖的体外降糖效果研究

为探究液体发酵茯苓胞外多糖的体外降糖效果,本研究以高胰岛素抵抗HepG2细胞为受试对象,分析了不同浓度的茯苓胞外多糖对葡萄糖消耗、细胞增殖和糖原合成的影响,并考察了茯苓胞外多糖对α-葡萄糖苷酶的抑制活性。研究结果显示,茯苓胞外多糖可显著促进高胰岛素抵抗HepG2细胞的葡萄糖消耗(p0.05),并加速其糖原合成,但对其细胞增殖无抑制活性;此外,茯苓胞外多糖在终浓度高于2.5μg/mL时还可表现出显著的α-葡萄糖苷酶抑制活性,最大抑制率可达29.51%。研究表明,液体发酵茯苓胞外多糖具有非常好的体外降糖效果。     

羧甲基茯苓多糖体外抗氧化活性研究

目的：比较不同浓度洗脱液洗脱得到的羧甲基茯苓多糖的抗氧化活性。方法：以茯苓为原料提取茯苓多糖,进行羧甲基取代反应,分离和纯化得到了均一性羧甲基茯苓多糖CMP-1、CMP-2、CMP-3、CMP-4,通过测定还原能力、DPPH自由基清除率、羟基自由基清除率、超氧阴离子自由基清除率比较其体外抗氧化活性。结果：羧甲基茯苓多糖均表现出与浓度正相关的体外抗氧化活性,其中CMP-4具有相对更强的体外抗氧化活性。结论：所得羧甲基茯苓多糖样品具有不同的体外抗氧化能力,随着洗脱液浓度增加,抗氧化活性增强。     

螺旋藻多糖对核酸内切酶活性和DNA修复合成的增强作用

本文用核酸内切酶实验和放射自显影术研究了螺旋藻水溶性多糖对DNA切除修复的效应。结果表明,该多糖能显著增强辐射引起DNA损伤的切除修复活性和程序外DNA合成(UDS)。考察切除修复的时程,发现螺旋藻多糖的存在不但能加快损伤DNA切除反应和UDS的初时速度,而且能延缓以上两个重要修复反应的饱和。     

发酵茯苓菌丝体和天然茯苓多糖的研究

本文对发酵茯苓菌丝体和天然茯苓中多糖的提取分离进行了研究,并分别测定了二者总多糖的提取率及总糖的平均含量。发酵茯苓菌丝体和天然茯苓中总多糖的提取率分别为24.47%和82.93%;发酵茯苓菌丝体和天然茯苓总糖的平均含量分别为45.17%和87.24%。     

倒卵叶五加多糖的乙酰化工艺研究

本文采用响应面法研究了倒卵叶五加多糖乙酰化的最优合成工艺,以多糖乙酰化的取代度为评价指标,考察了反应时间、乙酸酐与多糖的摩尔比和反应温度对多糖乙酰化取代度的影响。通过响应面法得到的最优工艺条件为:反应3.0 h,配料比3.7,反应温度70℃。初步的抗氧化性实验表明,乙酰化后的倒卵叶五加多糖的抗氧化性能有了明显提高。     

羧甲基茯苓多糖的制备及体内抗肿瘤作用的实验研究

目的 :探讨羧甲基茯苓多糖的制备方法及体内抗肿瘤作用。方法 :选择性氧化茯苓菌发酵液得羧甲基茯苓多糖 ,检测三个剂量的羧甲基茯苓多糖 (2 mg/ ml、4mg/ ml、6mg/ ml)对 H2 2 荷瘤小鼠淋巴细胞转化率和 NK细胞杀伤活性以及血清中 TNF-α含量的影响。结果 :羧甲基茯苓多糖三个剂量组均可提高荷瘤小鼠的淋巴细胞转化率和 NK细胞杀伤活性 ,与生理盐水组相比差异有显著性 (P<0 .0 5) ,中剂量组抗肿瘤的效果显著 ,与其他两组相比差异有显著性 (P<0 .0 5) ;羧甲基茯苓多糖三个剂量组均可提高荷瘤小鼠血清中 TNF-α的含量 ,与生理盐水组相比差异有显著性 (P<0 .0 5) ,但三个剂量组之间比较差异无显著性 (P<0 .0 5)。结论 :羧甲基茯苓多糖可改善荷瘤小鼠的免疫功能 ,具有抗肿瘤作用 ,且功效与作用剂量间有一定的关系 ,在最佳剂量时活性最高     

茯苓菌丝发酵液粗多糖对肠道益生菌的影响

目的探究茯苓菌丝发酵液中提取的粗多糖对肠道益生菌的影响。方法运用发酵工程相关技术获得茯苓菌丝发酵液,提纯分离以获取茯苓菌丝中分泌的粗多糖。将提取出的茯苓粗多糖以10.0%、5.0%、2.5%和空白对照(0.0%)的浓度对4种肠道益生菌——嗜酸乳杆菌、干酪乳杆菌、乳杆菌MZ900、青春双歧杆菌的生长进行观察。结果在茯苓粗多糖含量10.0%以下的范围内,10.0%、5.0%、2.5%浓度下肠道益生菌数量均高于对照组,并且随着茯苓发酵液粗多糖浓度的含量增加,4种肠道益生菌数量呈正相关增长。结论茯苓菌丝发酵液粗多糖对以上4种肠道益生菌有增菌效果。实验结果为茯苓菌丝发酵液粗多糖有保护人类肠道的作用提供理论依据,并为后续的生化研究与动物实验提供理论基础。     

款冬花多糖的羧甲基化衍生物合成工艺研究

本文应用响应面法优化制备款冬花多糖羧甲基化衍生物的合成工艺条件。采用碱性条件的氯乙酸法,以羧甲基的取代度为考察指标,最佳合成工艺为:氯乙酸浓度为2.5 mol/L,反应温度52℃,反应时间3.1 h。羧甲基化后的多糖清除DPPH的能力提高,抗氧化性提高,为今后多糖衍生物的产业化工艺开发有较好的参考价值和依据。     

茯苓多糖结构修饰及其与AA相互作用机理的研究

将茯苓多糖硫酸化得到硫酸化茯苓多糖(SP),应用光谱法研究了SP与天青A(AA)相互作用机理,并通过理论模型测得SP与AA最大结合数N=62,结合常数K=3.703×105。考察了反应体系中AA/SP摩尔比、NaCl、乙醇、羟丙基β环糊精以及TritonX 100对相互作用的影响。并对SP与AA相互作用机理提出了合理的解释。     

Structure characterization and carboxymethylation of arabinoxylan isolated from Ispaghula (Plantago ovata) seed husk

As revealed by NMR spectroscopy (after ultrasonic degradation) and HPLC (after total hydrolysis) an arabinoxylan (AX) containing 74.8% Xylp and 23.2% Araf was isolated from Ispaghula (Plantago ovata) by soaking the seed husk with water, extraction with aqueous sodium hydroxide and coagulation with acetic acid. The AX with a molar mass of 364,470 g/mol shows high swelling ability in water. The carboxymethylation of AX was carried out heterogeneously with sodium monochloroacetate in the presence of aqueous sodium hydroxide. The reaction parameters were varied in terms of slurry medium, molar ratio, temperature, time, and sodium hydroxide concentration. For comparative studies, carboxymethylation of arabinan was carried out. In order to determine the total degree of substitution (DS) and mole fractions of the repeating units of carboxymethyl arabinoxylan (CMAX) and of carboxymethyl arabinan, HPLC and ¹H NMR spectroscopic investigations after total hydrolysis were carried out. DS values for CMAX as high as 1.81 were achieved. CMAX is water soluble starting at DS of 0.33.     

Stepwise synthesis of oligopeptides with N-carboxy alpha-amino acid anhydrides

Oligopeptides were synthesized in high yields by the controlled coupling reaction of N-carboxy α-amino acid anhydrides (NCA's) with amino acid and peptide sodium salts in the heterogeneous reaction medium of acetonitrile–water which contained sodium carbonate. In this solvent, it could be considered that the reaction could occur at the interface of acetonitrile and aqueous layers, and that NCA's could be protected by the organic layer from side reactions such as hydrolysis and polymerization. The careful control of reaction conditions such as pH of the solution was not necessary when the heterogeneous mixed solvent was used. Sodium carbonate which had not been used for a reaction of this kind could be satisfactorily used for stabilizing the carbamate intermediates in the aqueous layer of the heterogeneous reaction medium.     

Preparation and characterization of cationic corn starch with a high degree of substitution in dioxane-THF-water media

Cationic corn starch derivatives with a high degree of substitution are prepared in alkaline solution or in mixed media of organic solvent and water with different levels of the cationic reagent, 2,3-epoxypropyltrimethylammonium chloride. The starch cationization yield is investigated, and the results indicate that the degree of substitution (DS) of the samples depends on the reaction conditions and reaction media. The maximum DS values are up to 1.37 in 1,4-dioxane alkaline-aqueous solution. Meanwhile, the structures of the cationic starch derivatives are characterized by elemental analyses, FTIR spectroscopy, X-ray diffraction, and ¹³C NMR spectroscopy, as well as by SEM techniques.     

高取代度CMS合成新工艺研究

研究以玉米淀粉为原料,用溶煤法合成CMS(2%水溶液,DS≥0.90,η≥120.0mpa.s)的新工艺;讨论了影响其合成的几种因素,确定了最佳技术参数和工艺条件,经工业性试验,证明本法切实可行。     

Single and multi-step carboxymethylation of water yam (Dioscorea alata) starch: synthesis and characterization

Water yam (Dioscorea alata) starch was carboxymethylated by single and multi-step processes for nine times successively. Optimal degree of substitution (DS) of 0.98 and reaction efficiency (RE) of 70.5% were obtained in the single-step carboxymethylation at 40 degrees C, 3 h in isopropanol-water reaction mixture ratio 1:0.16. The ratio of sodium hydroxide and sodium monochloroacetate moles to anhydroglucose unit moles for the optimal DS and RE were 1.62 and 1.39, respectively. The DS increased progressively as the steps of carboxymethylation increased from 2 to 9 and an optimal DS of 2.24 was obtained. Initial increases in carboxymethylation step increased the RE progressively up to 82.1% after the seventh carboxymethylation step but declined with further increases in the carboxymethylation step. The wide angle X-ray diffractometry and the DSC revealed remarkable changes after carboxymethylation because starch crystallinity reduced significantly. Thermogram of native starch showed a characteristic three-step decomposition with 13.16%, 61.54% and 24.79% weight losses progressively while carboxymethyl derivative showed four decomposition stages with 9.86%, 36.57%, 3.04% and 23.07% weight losses progressively. Studies revealed that carboxymethylation improved thermal stability of the native starch. IR spectrometry revealed that carboxymethyl starch showed new bands at nu=1600, 1426 and 1324 cm(-1). The broad band (13)C NMR spectra of the ultrasonically degraded carboxymethyl starch showed a peak at delta=180 ppm which was assigned to carbonyl carbon.     

Synthesis and characterization of a novel potato starch derivative with cationic acetylcholine groups

A novel substance, cationic acetylcholine potato starch (CAPS), was developed for the first time. The synthesis process had three steps: first, carboxymethyl potato starch (CMPS) was synthesized under sodium hydroxide alkaline condition and in isopropyl alcohol organic media; second, bromocholine chloride (BCC) was synthesized with sulphuric acid as a catalytic agent; finally, CAPS was synthesized by the reaction of CMPS with BCC in N,N'-dimethylformamide (DMF). The degree of substitution (DS) of CAPS was determined by ammonia gas-sensing electrode and elemental analysis. CAPS was characterized by Fourier transformed infrared (FTIR) and near infrared (FTNIR) spectroscopy, scanning electron microscopy (SEM), X-ray diffraction (XRD) and differential scanning calorimetry (DSC).     

Improving the industrial production of 6-APA: enzymatic hydrolysis of penicillin G in the presence of organic solvents

The hydrolysis of penicillin G in the presence of an organic solvent, used with the purpose of extracting it from the culture medium, may greatly simplify the industrial preparation of 6-APA. However, under these conditions, PGA immobilized onto Eupergit displays very low stability (half-life of 5 h in butanone-saturated water) and a significant degree of inhibition by the organic solvent (30%). The negative effect of the organic solvent strongly depended on the type of solvent utilized: water saturated with butanone (around 28% v/v) had a much more pronounced negative effect than that of methylisobutyl ketone (MIBK) (solubility in water was only 2%). These problems were sorted out by using a new penicillin G acylase derivative designed to work in the presence of organic solvents (with each enzyme molecule surrounded by an hydrophilic artificial environment) and a suitable organic solvent (MIBK). Using such solvent, this derivative kept its activity unaltered for 1 week at 32 degrees C. Moreover, the enzyme activity was hardly inhibited by the presence of the organic solvent. In this way, the new enzyme derivative thus prepared enables simplification of the industrial hydrolysis of penicillin G.     

羧甲基壳聚糖的制备及其在保鲜中的应用

研究了在碱性条件,无溶剂体系下壳聚糖羧甲基化制备工艺,通过IR对反应前后壳聚糖结构进行了表征。最佳反应条件：2g壳聚糖,在NaOH加入量为5g、碱化时间为6h、氯乙酸用量为6g、3％KI、反应时间为8h、反应温度60℃,产品取代度为1．27。同时研究了羧甲基壳聚糖对辣椒、猪肉涂膜保鲜实验,结果显示对辣椒、猪肉具有较好的涂膜保鲜作用。     

Preparation of carboxymethyl chitosan in aqueous solution under microwave irradiation

Carboxymethyl chitosan was prepared by reacting chitosan with chloroacetic acid in water under microwave irradiation. The effect of the reaction conditions was investigated and optimal conditions were identified. The influence of mass ratio of chloroacetic acid to chitosan, microwave power and pH on the degree of substitution or intrinsic viscosity were further studied. The degree of substitution of the carboxymethyl chitosan synthesized exceeded 0.85.     

Thermodynamic parameters monitoring the equilibrium shift of enzyme-catalyzed hydrolysis/synthesis reactions in favor of synthesis in mixtures of water and organic solvent

The main strategy developed to shift the equilibrium state of a hydrolase-catalyzed hydrolysis/synthesis reaction consists in reducing water activity by addition of organic solvents in the reaction medium. We have used several mixtures of water and 1,4-butanediol, ranging from pure water to pure 1,4-butanediol, to study the hydrolysis/synthesis reaction of the N-Cbz-L-tryptophanyl-glycineamide dipeptide, catalyzed by alpha-chymotrypsin. In the presence of 1,4-butanediol, alpha-chymotrypsin also catalyzed the esterification reaction between this diol and N-Cbz-L-tryptophan; this ester hydrolysis/synthesis reaction has thus also been examined. The dipeptide and ester equilibrium concentrations increase when the water content of the reaction medium is decreased. Using our experimental data, we have determined the equilibrium constants of the hydrolysis/synthesis equilibria involving the nonionized forms of the protected amino acids, the estimated values of which are Ksp = 8 10(5) for the dipeptide and Kse = 78 for the ester respectively. They are true thermodynamic equilibrium constants, each related to a single, well-defined reaction equilibrium and with water activity being taken into account. If an organic solvent is added to the reaction medium these equilibria can be shifted towards synthesis by decreasing the water activity but also by modifying the ionization/neutralization equilibrium constant of the ionizable groups. These two effects depend both on the water content and on the nature of the organic solvent used, and, in particular, on its dielectric constant. Because of the importance of this parameter in our study, we discuss using it as an indicator to select an appropriate organic solvent to perform an enzyme-catalyzed synthesis.