壳聚糖氧化降解制备聚合度6以下的壳寡糖

壳寡糖是壳聚糖经降解后的低聚物,其相对分子质量较小,水溶性好,易于吸收,生物活性高。本实验采用过氧化氢氧化降解脱乙酰度高于95%的壳聚糖,利用离子色谱-脉冲安培法对低聚合度的壳寡糖进行定性和定量分析,以制备聚合度在6以下的壳寡糖。以降解产物的离子色谱中聚合度6以下的峰面积和与产物得率之积作为响应指标,并结合凝胶渗透色谱来考察过氧化氢浓度、反应时间、反应温度对壳聚糖降解的影响及降解特性,并利用响应面分析法对氧化条件进行优化。研究结果表明,采用离子色谱测定聚合度6以下的壳寡糖在方法学上是可行的,具有良好的精密度、稳定性和重现性。得到的最佳降解工艺条件为:过氧化氢浓度4.50%、反应时间6 h、反应温度56℃。由最优条件下的降解产物离子色谱图可知,聚合度越低的壳寡糖越容易得到;可利用离子色谱法对壳聚糖的降解过程进行调控。     

壳聚糖氧化降解制备壳寡糖的研究

本文将脱乙酰度为96.7%的壳聚糖(COS)通过过氧化氢氧化降解制取壳寡糖,通过单因素变量的研究以及正交实验得出最优反应条件为:反应温度60℃,反应时间6 h,过氧化氢质量分数4.0%,乙酸质量分数4.0%,且四个因素对降解程度的影响为反应温度>过氧化氢质量分数>乙酸质量分数>反应时间。通过凝胶渗透色谱(GPC)对原料壳聚糖及最优条件下得到的降解产物的分子量分布进行检测,结果表明壳聚糖已完全降解且相对分子量达2000以下,降解产物经电喷雾质谱(ESI-MS)检测分析得聚合度为10以下,在控制降解的范围内,达到制备目的。     

过氧化氢法制备特定聚合度壳寡糖的工艺研究

实验对过氧化氢法氧化降解壳聚糖,制备聚合度为6～8的壳寡糖的工艺条件进行了研究。得到壳聚糖降解的最佳环境为弱酸性[3%HAc(v/v)],在料液比5%、反应时间8h、反应温度50℃、氧化剂浓度5%时,壳寡糖得率是62.70%,经GPC检测平均聚合度为6.87。聚合度符合预期要求,得率优于已报道的平均水平。     

低聚合度壳寡糖制备及其组分的色谱行为分析

利用壳聚糖酶水解壳聚糖,再通过乙醇沉淀法制备低聚合度(Dp＜6)的壳寡糖.利用Shodex NH2P-50 4E色谱柱,以乙腈-氨水为流动相梯度洗脱,建立了一种分离度好、灵敏度高的壳寡糖HPLC分析方法;利用UPLC-MS定性鉴定壳寡糖中N-乙酰化的组分.研究还探讨了壳寡糖组分的色谱行为,发现壳寡糖的N-乙酰化是导致壳寡糖不按照聚合度大小进行色谱分离的主要原因,为壳寡糖质量分析、组分分离提供了依据.     

酶膜耦合法制备高聚合度壳寡糖

为了制备高聚合度壳聚糖,本研究通过对比分批酶解法和酶膜耦合法制备所得壳寡糖产物的聚合度差异,探索了利用酶膜耦合技术高效富集制备高聚合度壳寡糖的可行性。研究结果表明,酶膜耦合方法所得壳寡糖产物中DP 4~8壳寡糖的总收率高达78.1%,DP 4~8壳寡糖所占比例分别为16.5%、35.8%、18.9%、7.81%和5.12%。同时,以卷式膜系统代替板式膜系统,通过错流过滤的方式,可以有效降低实验过程中的不可逆膜污染(R_if=1.56×106 m-1),提高了料液底物浓度(30 g/L)。综上所述,本研究建立了一种基于酶膜耦合技术的高聚合度壳寡糖连续制备工艺,为高聚合度壳寡糖的应用和功能研究提供了基础。     

聚合度为6～8的壳寡糖的制备

目的 研究聚合度6～8的壳寡糖的制备工艺.方法 选用合适的酶降解壳聚糖,超滤、干燥;用TLC、HPLC、MALDI-TOF-MS检测产品.结果 产品为白色,主要为聚合度6～8的壳寡糖.结论 此工艺能用于制备聚合度主要为6～8的壳寡糖.     

聚合度为6～8的壳寡糖的制备

目的研究聚合度6～8的壳寡糖的制备工艺。方法选用合适的酶降解壳聚糖,超滤、干燥;用TLC、HPLC、MALDI—TOF—MS检测产品。结果产品为白色,主要为聚合度6～8的壳寡糖。结论此工艺能用于制备聚合度主要为6～8的壳寡糖。     

南极磷虾壳聚糖、壳寡糖的制备与品质鉴定

本研究以南极磷虾壳为原料,制备较高品质的壳聚糖与壳寡糖,并对二者的品质进行鉴定。南极磷虾壳经脱钙、脱蛋白处理,探索脱乙酰反应条件（碱溶液浓度、反应温度与反应时间）,制备具有较高脱乙酰度的南极磷虾壳聚糖,并对壳聚糖的理化指标进行鉴定;探索酶法降解条件（壳聚糖酶添加量、酶解时间）,制备较高纯度的南极磷虾壳寡糖,并对壳寡糖的结构特征进行鉴定。结果表明,使用60%的氢氧化钠于110 ℃脱乙酰处理4 h制备的南极磷虾壳聚糖脱乙酰度为85.74%,粘均分子量为 305.65 kDa,水分含量4.66%,灰分含量0.98%,酸不溶物含量0.40%,各项理化指标均符合食品级壳聚糖的要求;使用壳聚糖酶水解南极磷虾壳聚糖制备壳寡糖,在壳聚糖酶添加量为0.2% （m/V）,酶解16 h条件下,南极磷虾壳寡糖产品得率为46.0%,红外光谱与NMR谱图显示了表征壳寡糖结构的全部特征峰,质谱结果显示南极磷虾壳寡糖主要由二糖(GlcN)₂、三糖(GlcN)₂-GlcNAc与四糖(GlcN)₃-GlcNAc构成。本研究通过制备较高品质的壳聚糖与壳寡糖,为南极磷虾壳的高值综合利用与南极磷虾新产品开发提供了技术支持。     

高效液相色谱法测定壳寡糖的含量

探讨高效液相色谱(HPLC)法在壳寡糖含量测定中的应用.选用Shodex Asahipak NH2P-50 4E色谱柱(4.6 mm×250 mm,5μm)测定壳寡糖样品中2～6个聚合度壳寡糖的含量.检测器为示差折光检测器,流动相V(乙腈):V(水)=75:25,流速1.2 mL/min,柱温30℃.液相检测壳寡糖的回收率为96.9％～98.2％,相对标准偏差为0.1％～1.0％.结果表明:该定量分析方法可快速、高效地测定壳寡糖样品中2～6个聚合度壳寡糖的含量.     

用于壳聚糖降解的复合酶制剂产生菌的筛选

以壳聚糖为唯一碳源,结合固态培养基发酵培养,从自然界中筛选得到一株产复合酶的菌株,初步鉴定为曲霉属.该菌产复合酶中以壳聚糖酶、纤维素酶和α-淀粉酶为主,它们的酶活分别为2.206、0.790、3.229U/mL.该复合酶降解壳聚糖的最适温度为50℃,最适pH为5.6,最适时间为4h.通过有机溶剂分级沉淀,发现该复合酶降解壳聚糖的最终产物以聚合度为8～16的壳寡糖为主,表明该复合酶在壳寡糖制备方面具有一定的应用价值.     

两种甲壳胺纤维的结构和性能比较

对2种甲壳胺纤维的结构和性能进行比较研究,发现不同企业生产的甲壳胺纤维的脱乙酰度、分子质量、结晶度、可染性和对双氧水的稳定性存在很大差别,脱乙酰度程度大和分子质量高的甲壳胺纤维具有更高的结晶度、更好的可染性和对双氧水的稳定性,在氧漂时的失重率更低。     

羊毛的过氧化氢—脱乙酰甲壳质防缩整理

采用双氧水和天然高分子聚合物壳聚糖对羊毛纤维进行防缩整理。研究了双氧水处理的pH值、时间、温度和浓度以及壳聚糖的分子量及其用量对防缩效果的影响。实验结果表明，采用这种方法处理后的羊毛在不明显损害其良好性能的基础上具有较好的防缩效果。     

低聚壳聚糖与葡萄糖和麦芽糖的美拉德反应及其衍生物的抗氧化性能

低聚壳聚糖(COS)作为氨基供体分别与提供羰基的葡萄糖和麦芽糖进行美拉德反应(氨基与羰基的物质量比均为3:1),考察了反应过程中吸光度和荧光值的变化。醇沉法提取低聚壳聚糖美拉德反应衍生物CG和CM。对两种衍生物进行红外表征和相对分子质量测定,并研究其对超氧阳离子O2.-、DPPH自由基的清除能力以及还原能力。结果显示:抗氧化能力强弱次序为CG>CM>COS,即美拉德反应后低聚壳聚糖衍生物抗氧化能力得到显著提高,且CG的抗氧化活性优于CM,表明与单糖进行美拉德反应制得的壳聚糖衍生物具有更好的抗氧化性。     

Efficacy of chitosan, carvacrol, and a hydrogen peroxide-based biocide against foodborne microorganisms in suspension and adhered to stainless steel

The ability of natural compounds to inactivate foodborne organisms adhered to surfaces was investigated with the ultimate aim of replacing synthetic biocides by more environmentally friendly, natural alternatives. The antimicrobial efficacy of 0.5, 1.0, and 2.0% chitosan and Spor-Klenz RTU (a commercial biocide based on hydrogen peroxide and peroxyacetic acid) and 0.5, 1.25, and 2.0 mM carvacrol was determined at 20 degrees C against Listeria monocytogenes, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, and Saccharomyces cerevisiae adhered to stainless steel disks. Treatment with up to 2.0% chitosan reduced the viable cell count in the microbial films of the four test organisms by 2.4, 1.8, 2.3, and 0.9 log CFU/test surface (t.s.), respectively. By contrast, planktonic counts of the same organisms were reduced by 0.8 to 1.7 log CFU/ml at 2.0% chitosan. Treatment with 2 mM carvacrol reduced the viable counts of adhered listeriae, salmonellae, and yeasts by 2 to 3 log CFU/t.s. but S. aureus counts were reduced by only 0.9 log CFU/t.s. The efficacy of any single compound was species specific. In the case of microbial films prepared using listeriae and salmonellae, Spor-Klenz RTU was most biocidal, followed by carvacrol and then chitosan. However, dried films of S. aureus were most sensitive to chitosan and relatively resistant to carvacrol and Spor-Klenz RTU. By contrast, yeast films were most sensitive to carvacrol and least sensitive to chitosan. It was concluded that carvacrol and chitosan may have potential for use as natural biocides although optimization of conditions would be necessary.     

H_2O_2/HAc氧化降解壳聚糖的探讨

采用过氧化氢-醋酸氧化降解法,探讨冰醋酸用量、反应温度、过氧化氢用量和反应时间对降解壳聚糖分子量的影响。     

木瓜蛋白酶-双氧水制备低分子量壳聚糖

该文对木瓜蛋白酶-双氧水降解壳聚糖的情况进行了研究,考察了温度、pH值、双氧水加入时间以及反应时间对壳聚糖分子量大小的影响,然后通过红外光谱分析壳聚糖降解产物的结构。结果表明:降解反应的最佳pH值为4,酶温度为45℃,双氧水反应温度为60℃,反应时间为24h;分子量降低导致晶格结构发生了变化,但是化学结构没有发生改变。     

猪小肠粘膜中肝素钠提取与精制工艺研究

以猪小肠粘膜为原料，进行肝素钠的提取与纯化，采用两种方法：一种是乙醇沉淀法，另一种是大孔阴离子树脂交换法。主要是通过树脂交换法提取肝素钠粗品，然后采用高锰酸钾和过氧化氢两步氧化法对粗品肝素进行除杂、提纯和精制，最后低温离心得到精品肝素钠。对传统的提取工艺条件进行了改进，并测得粗品平均收率为2．257％，精品平均收率为0．741％，其中粗品效价值为73．19U／mg，精品效价值为146．38U／mg。     

Preparation of antibacterial chito‐oligosaccharide by altering the degree of deacetylation of β‐chitosan in a Trichoderma harzianum chitinase‐hydrolysing process

BACKGROUND: Chito‐oligosaccharide (COS) is generally known to possess many specific biological functions, especially antibacterial activity, depending on its size. To prepare a specific size range of COS, however, has proved difficult. The aim of this study was to establish a method for preparing a specific size range of antibacterially active COS by adjusting the degree of deacetylation (DD) of β‐chitosan in a Trichoderma harzianum chitinase‐hydrolysing process. RESULTS: The molecular weight spectrum, elucidated by viscosity‐average molecular weight, high‐performance liquid chromatography and thin layer chromatography, of COS in chitosan hydrolysate was significantly related to the DD of its original chitosan. Compared with the original form, COS produced at 90% DD showed superior activity against most Gram‐negative bacteria tested, with a minimum inhibition concentration (MIC) ranging from 55 ± 27 to 200 ± 122 µ g mL^?1. Conversely, most Gram‐positive strains tested were less sensitive to COS (MIC > 880 ± 438 µ g mL^?1) than to its original form. Among the Gram‐positive strains, Staphylococcus xylosus was the only exception in that it showed a high susceptibility to COS and had an MIC as low as 45 ± 11 µ g mL^?1. CONCLUSION: The results indicate that the production of a specific size range of COS product is possible by altering the DD of chitosan in the chitinase‐catalysed process. To produce various sizes of COS for versatile biological functions, as seen in this study to inhibit various types of bacteria, is made possible in this established process. Copyright © 2008 Society of Chemical Industry     

Anticoagulant activity of heterochitosans and their oligosaccharide sulfates

Three kinds of partially deacetylated heterochitosans, 90% deacetylated chitosan, 75% deacetylated chitosan and 50% deacetylated chitosan, were prepared from crab chitin by N-deacetylation with 40% sodium hydroxide solution for different durations. Nine kinds of heterochitooligosaccharides (hetero-COSs) with relatively high molecular weights (5,000–10,000 Da; 90-HMWCOSs, 75-HMWCOSs, and 50-HMWCOSs), medium molecular weights (1,000–5,000 Da; 90-MMWCOSs, 75-MMWCOSs, and 50-MMWCOSs), and low molecular weights (below 1,000 Da; 90-LMWCOSs, 75-LMWCOSs, and 50-LMWCOSs) were prepared using an ultrafiltration membrane reactor system, respectively. In addition, their sulfated derivatives were prepared by a method using a trimethylamine-sulfur trioxide, and the anticoagulant properties of the heterochitosans and their COS sulfates with different chain lengths and degrees of deacetylation were investigated. Clotting times in thrombin-time assay were prolonged in the presence of various concentrations of the heterochitosans and their COS sulfates using normal human plasma. The 90% deacetylated chitosan sulfate exhibited the highest anticoagulant activity among all the heterochitosans and their COS sulfates.