Virion glycosylation governs integrity and infectivity of infectious pancreatic necrosis virus

The possible importance of the O-linked glycosylation in virion stability and infectivity of infectious pancreatic necrosis virus (IPNV) was analysed. Enzymatic treatment with O-glycosidase of radiolabelled virions under different ionic conditions, to allow for possible alternative exposure of glycosidic enzyme cleavage sites, did not alter the specific infectivity of virions re-isolated after rate-zonal centrifugation in glycerol gradients. As an alternative method to assess the significance of carbohydrates in IPNV integrity, periodate oxidation in the presence of an aldehyde quencher was chosen. Following re-isolation of viruses, a 3-5 (10)log-unit reduction in specific infectivity was revealed and, at higher concentrations, a total disruption or virion aggregation was observed. The loss of infectivity of intact virions was not because of a lack of attachment to cells. Additionally, re-evaluation of reading values from UV-spectra of purified IPNV yielded a specific infectivity of 3 × 10(11) TCID(50)-units mg(-1) of protein and a ratio of 40 virions per TCID(50)-unit in the CHSE-214 cell system.     

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.     

洋葱苯丙氨酸解氨酶基因AcPAL2的克隆与表达分析

利用简并PCR技术和RACE技术克隆得到了一条洋葱的苯丙氨酸解氨酶(PAL)基因全长cDNA序列。该cDNA序列全长2349 bp,编码长685个氨基酸残基的多肽序列,命名为AcPAL2。Blast分析表明该序列与虎眼万年青Galtonia saundersiae、野蕉Musa balbisiana的相似性均较高。Real-time PCR表达及花青素含量分析表明,红皮洋葱该基因表达量最大,而黄皮和白皮洋葱表达量极低;在红皮洋葱中该基因在膨大初期大量表达,并迅速降低至一定程度后趋于相对平稳表达,且与花青素的积累过程相一致。     

Glycoproteomic profiling of serum peptides in canine lymphoma and transitional cell carcinoma

Differential expression of fucosylated glycoproteins has been correlated with malignancy and metastatic potential in various types of neoplasia. Utilizing glycoproteomics techniques, changes in fucosylated serum peptides associated with naturally occurring canine lymphoma and transitional cell carcinoma (TCC) have been evaluated. In both types of neoplasia, the majority of the fucosylated peptides that changed increased with the cancer. In one lymphoma case that was examined over the course of the disease, the same fucosylated peptides that increased during pre-chemotherapy decreased during post-chemotherapy, and then subsequently increased upon recurrence of the lymphoma. When comparing all the fucosylated peptides that increased in both types of cancer, there were only two peptides in common allowing discrimination between lymphoma and TCC based on their peptide profiles. These results emphasize the prospect of glycopeptide profiling in proteomics for use in discovering a panel of non-invasive, diagnostic or prognostic biomarkers of cancer.     

Surface Glucan Structures in Aeromonas spp.

Aeromonas spp. are generally found in aquatic environments, although they have also been isolated from both fresh and processed food. These Gram-negative, rod-shaped bacteria are mostly infective to poikilothermic animals, although they are also considered opportunistic pathogens of both aquatic and terrestrial homeotherms, and some species have been associated with gastrointestinal and extraintestinal septicemic infections in humans. Among the different pathogenic factors associated with virulence, several cell-surface glucans have been shown to contribute to colonization and survival of Aeromonas pathogenic strains, in different hosts. Lipopolysaccharide (LPS), capsule and α-glucan structures, for instance, have been shown to play important roles in bacterial–host interactions related to pathogenesis, such as adherence, biofilm formation, or immune evasion. In addition, glycosylation of both polar and lateral flagella has been shown to be mandatory for flagella production and motility in different Aeromonas strains, and has also been associated with increased bacterial adhesion, biofilm formation, and induction of the host proinflammatory response. The main aspects of these structures are covered in this review.     

乳清蛋白-低聚异麦芽糖的制备及抗原性研究

将低聚异麦芽糖通过糖基化引入乳清蛋白制备乳清蛋白-低聚异麦芽糖,用间接竞争ELISA法测定不同乳清蛋白与低聚异麦芽糖质量比,不同反应时间生成的乳清蛋白-低聚异麦芽糖中α-乳白蛋白和β-乳球蛋白抗原性的变化。结果表明:糖基化能有效降低乳清蛋白中α-乳白蛋白和β-乳球蛋白的抗原性;不同反应时间、不同乳清蛋白与低聚异麦芽糖质量比对乳清蛋白中α-乳白蛋白和β-乳球蛋白抗原性的影响不同,乳清蛋白与低聚异麦芽糖的质量比为1∶4,反应24 h时效果最好,α-乳白蛋白的抗原性从25.67μg/mL降低到9.33μg/mL,β-乳球蛋白的抗原性从97.82μg/mL降低到28.25μg/mL。     

糖基化鸡蛋清大豆复合蛋白饮料加工技术研究

研究糖基化鸡蛋清大豆复合蛋白饮料的加工工艺,通过单因素及正交试验,确定了最佳配方及工艺条件。试验结果表明,选择合适的复合乳化稳定剂和采用预乳化及二次高压均质工艺,均有助于提高复合蛋白饮料的稳定性。     

马传染性贫血病毒囊膜基因gp90V3区糖化回复突变感染性克隆的构建

为阐明我国马传染性贫血病毒（Equine infectious anemia virus,EIAV）弱毒疫苗致弱及免疫保护的分子机制,作者分析了疫苗株及其亲本强毒株共34个囊膜基因序列。结果发现疫苗株在膜基因gp90V3区的潜在N-连接糖基化位点出现稳定的碱基替换,使该位点消失。为研究囊膜糖基化的作用,以疫苗株全长感染性克隆pLG—FD3—8为亲本,利用反向遗传技术对该突变位点进行糖基化序列回复操作,构建全基因感染性克隆pLGFDg5。将pLGFDg5转染驴胎皮肤细胞（FDD）,通过逆转录酶活性和RT-PCR方法评价其感染性。结果表明,将pLGFDg5在FDD细胞中盲传3代后,可在细胞培养上清中检测到逆转录酶活性,用RT—PCR检测到EIAV保守基因片段,在电镜下可见典型的EIAV粒子。在FDD细胞上的病毒复制动力学分析显示,与其亲本克隆pLGFD3—8的衍生病毒pLGFD3-V相比,回复突变克隆衍生的pLGFDg5-V复制速度较慢,获得较低的病毒载量。体外抗体中和试验表明,与其亲本pLGFD3-V相比,引入潜在的糖基化位点g5降低了衍生病毒对中和抗体的敏感性。这一结果为N-连接糖基化在我国马传贫弱毒疫苗致弱机理的作用研究提供了重要依据。     

Evidence for the Non-Glycoprotein Nature of High Molecular Weight Glutenin Subunits of Wheat

An isolation and purification procedure for wheat high molecular weight glutenin subunits (HMW-GS) involving no steps expected to cause deglycosylation of glycoproteins was developed to allow removal of the maximum amount of carbohydrates not covalently linked to HMW-GS. Flour was defatted and then extracted with 50%n-propanol (50%n-PrOH) to remove arabinogalactans and glucose-containing material. HMW-GS were extracted with 50%n-PrOH containing 5% β-mercaptoethanol, precipitated in 60%n-PrOH, alkylated and recovered by precipitation. The partially purified HMW-GS were separated by cation exchange chromatography and resulting fractions purified by RP–HPLC. The purified HMW-GS contained only 0·1% of glucose (corresponding to 51–59 monosaccharide residues per 100 HMW-GS molecules) and no other amino or neutral sugars. The HMW-GS gave a positive reaction in a periodate–digoxigenin glycan detection assay when labelled in solution (with or without periodate oxidation) but not when labelled directly on-the-blot. HMW-GS expressed inEscherichia coli, a micro-organism without glycosylation capabilities, reacted in an identical way. Possible reasons are given for the difference in results obtained after labelling in solution or on-the-blot. No positive reaction between the purified HMW-GS and mannose-specificGalanthus nivalisagglutinin was observed. The presence of N-acetylglucosamine in HMW-GS has been previously reported²⁰. Since neither mannose nor N-acetylgalactosamine were found in the HMW-GS, and these proteins lack the sequon necessary for N-glycosylation, only O-linked GlcNAc moieties are possible. This modification could not be detected using a galactosyltransferase labelling assay. Taken together these results suggest that HMW-GS are not glycosylated.     

葡萄中糖基化花色苷研究进展

葡萄果实与葡萄酒的颜色由其所含花色苷的种类和含量决定,而花色苷是由花色素经过糖基化修饰转变而成,所以糖基化修饰在葡萄果实花色苷合成途径中起着重要作用。葡萄果实中的糖基化花色苷主要包括花色素的3–O–葡萄糖基和3,5–O–双葡萄糖基,即花色素单糖苷和花色素双糖苷,糖基化花色苷的组成是决定红葡萄酒品质的关键因素之一。对糖基化花色苷在葡萄果实中的组成及其对葡萄酒颜色和稳定性的影响进行了简要介绍,重点对花色素单糖苷和花色素双糖苷合成的关键酶基因以及转录因子进行了综述,以期为葡萄果实糖基化花色苷合成的调控机理的全面揭示和优质红色酿酒葡萄品种的选育提供信息。