壳寡糖对凡纳滨对虾生长和免疫力的影响

为研究壳寡糖（ch itooligosaccharides,COS）对凡纳滨对虾（Litopemaeus vannam ei）生长和非特异性免疫水平的影响,设置了含COS分别为0（对照）、250 mg.kg-1、500 mg.kg-1、750 mg.kg-1和1 000 mg.kg-1的5组试验饲料,进行了4周的饲养试验。结果显示,COS添加量为250 mg.kg-1和500 mg.kg-1时凡纳滨对虾血细胞总数、生长率和成活率显著提高（P〈0.05）,含COS 250 mg.kg-1时对虾血清中的过氧化物酶（peroxidase,POD）活力显著提高（P〈0.05）,添加量为500 mg.kg-1时酸性磷酸酶（ac id phosphatase,ACP）和溶菌酶（lysozym e,LSZ）活力显著提高（P〈0.05）,添加量为250 mg.kg-1和500 mg.kg-1时总超氧化物歧化酶（total superoxide d ismutase,T-SOD）活力提高不显著（P〉0.05）。由此表明饲料中添加250～500 mg.kg-1COS可使凡纳滨对虾细胞免疫和体液免疫因子总体活力达到较高水平,可显著提高凡纳滨对虾的生长率和成活率。     

Bioproduction of Chitooligosaccharides: Present and Perspectives

Chitin and chitosan oligosaccharides (COS) have been traditionally obtained by chemical digestion with strong acids. In light of the difficulties associated with these traditional production processes, environmentally compatible and reproducible production alternatives are desirable. Unlike chemical digestion, biodegradation of chitin and chitosan by enzymes or microorganisms does not require the use of toxic chemicals or excessive amounts of wastewater. Enzyme preparations with chitinase, chitosanase, and lysozymeare primarily used to hydrolyze chitin and chitosan. Commercial preparations of cellulase, protease, lipase, and pepsin provide another opportunity for oligosaccharide production. In addition to their hydrolytic activities, the transglycosylation activity of chitinolytic enzymes might be exploited for the synthesis of desired chitin oligomers and their derivatives. Chitin deacetylase is also potentially useful for the preparation of oligosaccharides. Recently, direct production of oligosaccharides from chitin and crab shells by a combination of mechanochemical grinding and enzymatic hydrolysis has been reported. Together with these, other emerging technologies such as direct degradation of chitin from crustacean shells and microbial cell walls, enzymatic synthesis of COS from small building blocks, and protein engineering technology for chitin-related enzymes have been discussed as the most significant challenge for industrial application.     

低分子质量壳寡糖对蛋鸡生产性能、蛋品质、血清生化指标、盲肠微生物数量及脾脏白细胞介素-2、肿瘤坏死因子-α基因表达的影响

本试验旨在研究饲粮中添加不同水平的低分子质量(1 000 u)壳寡糖对蛋鸡生产性能、蛋品质、血清生化指标、盲肠微生物数量以及脾脏白细胞介素-2(IL-2)和肿瘤坏死因子-α(TNF-α)基因表达的影响。选取体重和产蛋率相近的58周龄海兰褐壳蛋鸡600只,随机分为4组,每组5个重复,每个重复30只。对照组饲喂基础饲粮,试验组分别饲喂在基础饲粮中添加300、600、900 mg/kg壳寡糖的试验饲粮。预试期为7 d,正试期为42 d。结果表明:1)添加300、600和900 mg/kg壳寡糖组的产蛋率分别比对照组提高了4.52%(P0.05)、2.99%(P0.05)和4.08%(P0.05)。2)试验第3、6周末,添加600和900 mg/kg壳寡糖组的鸡蛋哈夫单位分别比对照组提高了6.87%、6.69%和6.47%、6.60%(P0.05)。3)与对照组相比,饲粮添加600、900 mg/kg壳寡糖显著降低了血清葡萄糖、胆固醇含量和谷草转氨酶活性(P0.05)。4)与对照组相比,饲粮添加600、900 mg/kg壳寡糖显著提高了盲肠双歧杆菌和乳酸杆菌的数量(P0.05),显著降低了盲肠金黄色葡萄球菌的数量(P0.05)。5)与对照组相比,饲粮添加300、600 mg/kg壳寡糖显著提高了脾脏IL-2 m RNA表达水平(P0.05),饲粮添加600 mg/kg壳寡糖显著提高了脾脏TNF-αm RNA表达水平(P0.05)。由此可见,饲粮中添加不同水平的壳寡糖,提高了蛋鸡的产蛋率和哈夫单位,调节了肠道微生物菌群,增强了蛋鸡的免疫力,适宜壳寡糖添加水平为600 mg/kg。     

Chitin Deacetylases: Properties and Applications

Chitin deacetylases, occurring in marine bacteria, several fungi and a few insects, catalyze the deacetylation of chitin, a structural biopolymer found in countless forms of marine life, fungal cell and spore walls as well as insect cuticle and peritrophic matrices. The deacetylases recognize a sequence of four GlcNAc units in the substrate, one of which undergoes deacetylation: the resulting chitosan has a more regular deacetylation pattern than a chitosan treated with hot NaOH. Nevertheless plain chitin is a poor substrate, but glycolated, reprecipitated or depolymerized chitins are good ones. The marine Vibrio sp. colonize the chitin particles and decompose the chitin thanks to the concerted action of chitinases and deacetylases, otherwise they could not tolerate chitosan, a recognized antibacterial biopolymer. In fact, chitosan is used to prevent infections in fishes and crustaceans. Considering that chitin deacetylases play very important roles in the biological attack and defense systems, they may find applications for the biological control of fungal plant pathogens or insect pests in agriculture and for the biocontrol of opportunistic fungal human pathogens.     

Anti-Inflammatory Activity of Chitooligosaccharides in Vivo

All the reports to date on the anti-inflammatory activity of chitooligosaccharides (COS) are mostly based on in vitro methods. In this work, the anti-inflammatory activity of two COS mixtures is characterized in vivo (using balb/c mice), following the carrageenan-induced paw edema method. This is a widely accepted animal model of acute inflammation to evaluate the anti-inflammatory effect of drugs. Our data suggest that COS possess anti-inflammatory activity, which is dependent on dose and, at higher doses, also on the molecular weight. A single dose of 500 mg/kg b.w. weight may be suitable to treat acute inflammation cases; however, further studies are needed to ascertain the effect upon longer inflammation periods as well as studies upon the bioavailability of these compounds.     

黄粉虫水溶性低聚糖的提取及其对大肠杆菌抑菌作用研究

为开发昆虫几丁质资源,扩大其应用范围,本试验采用酸碱法从烘干的黄粉虫幼虫体内提取几丁质粗品,再经碱处理得到壳聚糖,壳聚糖经过适宜的温度和过氧化氢浓度处理后得到水溶性的低聚糖。黄粉虫水溶性低聚糖的浓度为0.06～0.3 mg.mL~(-1)对大肠杆菌均有很强的抑制效果。浓度为0.12、0.18、0.24、0.3 mg.mL~(-1)抑制作用均达到极显著水平,浓度为0.06 mg.mL~(-1)达到显著水平。利用黄粉虫提取的水溶性低聚糖,具有较好的溶解性能,在一定浓度下对大肠杆菌有较强的抑菌作用。     

酶法制备壳寡糖研究现状

对不同酶法制备壳寡糖的研究现状与进展进行了综述,这些酶分为专一性酶和非专一性酶.专一性酶介绍了壳聚糖酶、几丁质酶及溶菌酶,非专一性酶介绍了纤维素酶及混合酶,并将这些酶的部分性质与作用机理也进行简单介绍.     

壳寡糖对三疣梭子蟹免疫力的影响

为研究壳寡糖对三疣梭子蟹免疫功能的影响,设置了含壳寡糖0.0、0.5、1.0、1.5和2.0 g/kg（分别为T0、T1、T2、T3和T4组）的5组试验饲料饲喂三疣梭子蟹,分别在第10 d、20 d和30 d取血清进行酚氧化酶（PO）、酸性磷酸酶（ACP）、碱性磷酸酶（AKP）、总超氧化物歧化酶（T-SOD）、溶菌酶（LSZ）和过氧化物酶（POD）活性的测定,试验结束时测定血细胞的密度。结果显示,在基础饲料中添加适量的壳寡糖可以显著性提高ACP、T-SOD、LSZ、POD活力（P〈0.05）,对PO和AKP活力促进效果不显著（P〉0.05）。壳寡糖可以显著提高三疣梭子蟹血细胞密度（P〈0.05）,对3种血细胞所占比例有一定的影响（P〉0.05）。综合考虑,T2、T3组对三疣梭子蟹免疫促进效果较好,建议基础饲料中壳寡糖添加量为1.0～1.5 g/kg。     

壳寡糖对小鼠胚胎体外发育的影响

本研究旨在通过向小鼠体内受精和体外受精原核胚胎的培养基中添加壳寡糖(对照组:0μg·m L-1;试验组:5、10、50、100和200μg·m L-1),研究其对小鼠胚胎着床前发育效率(卵裂率、囊胚率)和囊胚质量(囊胚总细胞数、内细胞团数(ICM)与总细胞数的比率)的影响。结果表明,对于自然受精胚胎,添加10μg·m L-1壳寡糖组的囊胚总细胞数显著高于对照组(P0.05),添加200μg·m L-1壳寡糖组的卵裂率和囊胚率均显著低于对照组(P0.05)。对于体外受精胚胎,添加10μg·m L-1壳寡糖组的囊胚率显著高于对照组(P0.05),添加200μg·m L-1壳寡糖组的囊胚率显著低于对照组(P0.05)。本试验表明,在化学限定培养基中添加适量的壳寡糖有利于改善小鼠胚胎的质量,但高剂量的壳寡糖可能不利于小鼠胚胎的早期发育。     

6-羧基壳寡糖螯合钙制备工艺与络合性能研究

以壳寡糖(COS)为试验材料,采用TEMPO氧化法将壳寡糖C6位上的羟甲基直接氧化成为羧基,制备6-羧基壳寡糖(CCOS)。以Ca Cl2作为钙源,通过探讨体系pH值、反应温度、反应时间、CCOS与Ca Cl2质量比对CCOS-Ca(Ⅱ)得率的影响,采用三因素五水平中心旋转组合设计优化了制备工艺。并对产物进行傅里叶变换红外光谱扫描及元素分析。使用Design-Expert软件对响应面试验数据进行回归分析。所得CCOS-Ca(Ⅱ)制备最佳工艺参数为:反应pH值为8.3,反应时间60 min,反应温度50℃,CCOS与Ca Cl2质量比为34。在此优化工艺条件下,验证试验螯合率为88.86%。通过红外光谱及元素分析,证明了钙螯合物的形成,CCOS-Ca(Ⅱ)配合物的结构初步认为CCOS与Ca2+摩尔比为2∶1。CCOS分子除了含有—OH、—NH2外,还含有—COOH,能够对钙进行有效螯合,体系pH值、反应温度、配体质量比对螯合结果影响显著,所得CCOS-Ca(Ⅱ)络合物不仅能发挥壳寡糖本身独特的生物活性,而且有可能起到协同效应,增强各自的生物活性。