Evidence for a Negative Correlation between Human Reactive Enamine-Imine Intermediate Deaminase A (RIDA) Activity and Cell Proliferation Rate: Role of Lysine Succinylation of RIDA

Reactive intermediate deaminase (Rid) proteins are enzymes conserved in all domains of life. UK114, a mammalian member of RidA subfamily, has been firstly identified as a component of liver perchloric acid-soluble proteins (L-PSP). Although still poorly defined, several functions have been attributed to the mammalian protein UK114/RIDA, including the reactive intermediate deamination activity. The expression of UK114/RIDA has been observed in some tumors, arousing interest in this protein as an evaluable tumor marker. However, other studies reported a negative correlation between UK114/RIDA expression, tumor differentiation degree and cell proliferation. This work addressed the question of UK114/RIDA expression in human non-tumor HEK293 cell lines and in some human tumor cell lines. Here we reported that human RIDA (hRIDA) was expressed in all the analyzed cell line and subjected to lysine (K-)succinylation. In HEK293, hRIDA K-succinylation was negatively correlated to the cell proliferation rate and was under the control of SIRT5. Moreover, K-succinylation clearly altered hRIDA quantification by immunoblotting, explaining, at least in part, some discrepancies about RIDA expression reported in previous studies. We found that hRIDA was able to deaminate reactive enamine-imine intermediates and that K-succinylation drastically reduced deaminase activity. As predicted by in silico analysis, the observed reduction of deaminase activity has been related to the drastic alterations of hRIDA structure inferred by K-succinylation. The role of hRIDA and the importance of its K-succinylation in cell metabolism, especially in cancer biology, have been discussed.     

琥珀酸酐改性麦草浆对Cd(Ⅱ)的吸附性能

以二甲苯作溶剂、三乙胺作催化剂,通过琥珀酸酐改性麦草浆(WSP),制备出一种新型吸附剂S-WSP。通过红外光谱对S-WSP进行表征结构分析,该酯化反应在异相催化条件下能够顺利进行。S-WSP的羧酸基团与NaOH溶液作用去质子化可成功获得NaS-WSP。NaS-WSP悬浮在水溶液中吸附Cd2+,最大吸附量可达105.7mg/g,其动力学过程遵循伪二级动力学模型。由于在离子交换过程中竞争质子,该吸附剂的吸附能力会随着pH的下降而降低。     

琥珀酰化度大小对改性小麦面筋蛋白性质影响的研究

本文采用琥珀酸酐对小麦面筋蛋白进行酰化改性，讨论了琥珀酰化度大小对面筋蛋白的溶解性、乳化性、乳化稳定性、起泡性和起泡稳定性影响，并对最佳改性条件下改性小麦面筋蛋白进行了在面条加工中的应用研究。结果表明：琥珀酰化度为66．1％时改性效果最好，改性面筋蛋白的溶解度、乳化及乳化稳定性、起泡及起泡稳定性分别为5．09mg／ml，56．8％，56．4％，44．8％，25％；利用最佳改性条件下的琥珀酰化面筋蛋白生产面条，面条的综合性能均比未改性的面筋蛋白品质高。     

琥珀酰化处理对芸豆蛋白乳化性和体外消化性的影响

本文研究了琥珀酰化对芸豆分离蛋白（KPI）乳化性和体外消化性的影响。KPI酰化包括两个主要的阶段,酸酐-蛋白比为0-0.2．2,为ε-氨基（Lys）酰化阶段（N-酰化）;再增加酸酐与蛋白比,ε-氨基酰化基本完成,反应进入羟基（Thr,Ser）酰化阶段（O-酰化）。在低离子强度下,蛋白的EAI和ESI随N-酰化度增加急剧增加,而O-酰化基本不影响蛋白的EAI和ESI;对照和酰化KPI的EAI和ESI均随溶剂体系离子强度增加而降低。酰化改善KPI的体外消化性能。     

相同条件下不同酸酐/蛋白酰化程度与谱学行为的研究

目的:研究大豆分离蛋白(SPI)经酰化改性后的不同酰化程度与蛋白谱学行为。方法:采用紫外光谱和红外光谱探讨酰化对蛋白分子结构特征的影响。结果:紫外光谱分析显示,酰化处理后SPI肽链展开;红外光谱显示,酸酐首先选择蛋白质赖氨酸上ε-NH2残基反应,当ε-NH2残基被大量修饰后,赖氨酸、丝氨酸和苏氨酸的羟基的酰基修饰程度增加。另一方面,蛋白多肽链的二级结构发生较大变化,即SPI中二级结构由β-折叠为主变为以无规则卷曲为主。结论:实验结果有助于阐明SPI酰化后功能性质发生变化的内在结构因素。     

Effect of gelatinisation on slowly digestible starch and resistant starch of heat-moisture treated and chemically modified canna starches

The effects of gelatinisation on slowly digestible (SDS) and resistant starch (RS) of native and modified canna starches were investigated. Starch slurries (10% w/w) were gelatinised at 100 °C for 5, 10, 20 and 40 min using a rapid visco analyzer (RVA). Significant change in the degree of gelatinisation (DG) values of all starch samples was observed during the initial 10 min of gelatinisation; after that the DG values increased gradually with gelatinisation time. The RS contents in all gelatinised starches decreased with increasing gelatinisation time, while the SDS values fluctuated. Chemical modification affected DG values as well as RS/SDS contents. The RS contents in 10% (w/w) acetylated, hydroxypropylated, octenyl succinylated and cross-linked canna starches gelatinised at 100 °C for 40 min were 26.6%, 32.0%, 45.3% and 19.8%, respectively, which were higher than that of the native starch (12.4%). Canna starch modified by crosslinking had the highest SDS content when gelatinised for 20-40 min. Modification of canna starch by heat-moisture treatment resulted in a lower content of RS for all treated samples. However, the Vt-HMT25 (canna starch containing moisture content of 25% during heat treatment) when gelatinised for 5-20 min contained a higher amount of SDS, compared to unmodified starch. The most effective modification method for RS and SDS formation was octenyl succinylation, where the sum of RS and SDS approached that of Novelose260.     

Structure,Biosynthesis, and Biological Activity of Succinylated Forms of Bacteriocin BacSp222

BacSp222 is a multifunctional peptide produced by Staphylococcus pseudintermedius 222. This 50-amino acid long peptide belongs to subclass IId of bacteriocins and forms a four-helix bundle molecule. In addition to bactericidal functions, BacSp222 possesses also features of a virulence factor, manifested in immunomodulatory and cytotoxic activities toward eukaryotic cells. In the present study, we demonstrate that BacSp222 is produced in several post-translationally modified forms, succinylated at the ε-amino group of lysine residues. Such modifications have not been previously described for any bacteriocins. NMR and circular dichroism spectroscopy studies have shown that the modifications do not alter the spatial structure of the peptide. At the same time, succinylation significantly diminishes its bactericidal and cytotoxic potential. We demonstrate that the modification of the bacteriocin is an effect of non-enzymatic reaction with a highly reactive intracellular metabolite, i.e., succinyl-coenzyme A. The production of succinylated forms of the bacteriocin depends on environmental factors and on the access of bacteria to nutrients. Our study indicates that the production of succinylated forms of bacteriocin occurs in response to the changing environment, protects producer cells against the autotoxicity of the excreted peptide, and limits the pathogenicity of the strain.     

小麦面筋蛋白琥珀酰化修饰研究

采用琥珀酸酐对小麦面筋蛋白进行改性修饰，系统研究了底物浓度、改性剂用量和反应温度对反应程度、溶解性、乳化性、起泡性和泡沫稳定性等蛋白功能特性的影响。优化出小麦面筋蛋白琥珀酰化的最佳条件：琥珀酰化底物浓度10％，琥珀酸酐用量15％，反应温度45℃。应用红外光谱对最优条件下改性的面筋蛋白结构进行了表征。将琥珀酰化改性后的小麦面筋蛋白添加至小麦粉中，面团粘着性提高32％。     

Physicochemical Properties of Acylated Casein Micelles in Milk

ABSTRACT: The objective of this study was to determine the effects of acetylation or succinylation on the physicochemical properties of casein micelles in milk. Analysis of untreated and modified milk samples revealed solubilization of caseins and calcium phosphate from the micelle under mild acylation treatments. Solvation of micelle was affected, whereas no influence in size dia and potential zeta arose from the acylation treatments. Physicochemical characteristics of acetylated milk sample were intermediate between untreated and succinylated ones. Changes were also observed after calcium addition to reconstituted milk. Calcium supplementation counteracted the effect of chemical modification of caseins by reducing the susceptibility of casein micelle to disassembly.