Labeling and Characterization of Phenol-Containing Glycopeptides Using Chemoselective Probes with Isotope Tags

Secondary metabolites are structurally diverse natural products (NPs) and have been widely used for medical applications. Developing new tools to enrich NPs can be a promising solution to isolate novel NPs from the native and complex samples. Here, we developed native and deuterated chemoselective labeling probes to target phenol-containing glycopeptides by the ene-type labeling used in proteomic research. The clickable azido-linker was included for further biotin functionalization to facilitate the enrichment of labeled substrates. Afterward, our chemoselective method, in conjunction with LC-MS and MSn analysis, was demonstrated in bacterial cultures. A vancomycin-related phenol-containing glycopeptide was labeled and characterized by our labeling strategy, showing its potential in glycopeptide discovery in complex environments.     

基于核磁共振代谢组学分析不同品种草莓的化学成分差异

目的采用基于核磁共振(nuclear magnetic resonance,NMR)的代谢组学方法对"丰香""久香""宁玉"和"甜查理"4个不同品种草莓进行指纹图谱比较和化学成分差异分析。方法采用主成分分析法综合评价4个不同品种草莓的整体分布状况;采用正交偏最小二乘判别分析法筛选并鉴别出引起不同草莓品种有所差异的潜在差异代谢物。结果研究发现,苹果酸在"丰香"草莓中含量最低,蔗糖含量则最高;蔗糖在"久香"草莓中的含量最低;丙氨酸、柠檬酸、天冬酰胺、D-葡萄糖和D-果糖在"宁玉"中含量最低;丙氨酸、苹果酸、柠檬酸、天冬氨酸和D-葡萄糖在"甜查理"中含量最高。结论基于NMR的代谢组学技术可以用于草莓品种的鉴别和化学成分的差异分析,并为草莓的品种溯源提供科学数据。     

Identification of factors that accelerate hydrogen production by Clostridium butyricum RAK25832 using casamino acids as a nitrogen source

The ability of Clostridium butyricum RAK25832 to use casamino acids as a nitrogen source was investigated. Strain RAK25832 showed the capacity to utilize different types of carbon sources. With glucose as a carbon source (10 g/L), the preferred final concentration of casamino acids was 26.67 g/L, with a cumulative hydrogen production, production rate, and yield of 2505 mL H₂/L, 160 mL/h, and 1.81 mol H₂/mol glucose, respectively. Eighteen metal elements were screened to identify the most important metals for biohydrogen production, and four elements were optimized. The optimal medium composition was MgCl₂·6H₂O (0.1 g/L), K₂HPO₄·3H₂O (6.67 g/L), NaHCO₃ (2.6 g/L), and FeCl₂·4H₂O (0.002 g/L). Vitamin supplementation of the medium showed no significant effect on hydrogen production. Under the optimized conditions, cumulative hydrogen production reached 3074 mL H₂/L. This is the first study to demonstrate the use of casamino acids as a nitrogen source by C. butyricum.     

氰霜唑及其代谢物4-氯-5-(4-甲苯基)-1H-咪唑-2-腈在苦瓜中的残留消解动态及膳食安全性评价

[目的]研究旨在优化苦瓜中氰霜唑(Cyazofamid)及其代谢物CCIM的QuEChERS-液相色谱-串联质谱残留分析方法；同时结合苦瓜的田间残留消解动态试验,对膳食摄入风险进行评价。[方法]苦瓜样品采用含1%乙酸的乙腈溶液提取,LC-MS/MS进行检测,通过基质匹配标准溶液外标法定量。通过在黑龙江、河北、河南、重庆、浙江和广东6地进行了100 g/L氰霜唑悬浮剂在苦瓜上残留的田间试验,研究了氰霜唑在苦瓜中的消解动态,对苦瓜中氰霜唑可能产生的膳食摄入风险安全进行评价。[结果]试验结果表明：1)氰霜唑在0.005-1.0 mg/L、CCIM在0.001-1.0 mg/L质量浓度范围内线性关系良好,相关系数均大于0.9987；在0.010、0.10、0.50 mg/kg 3个浓度下,苦瓜中氰霜唑及CCIM的回收率为80.0%～100.0%,相对标准偏差(RSD)为0.05%～12.1%。检出限(LOD)为0.364-0.654 μg/kg,方法定量限(LOQ)为添加的最低浓度0.010 mg/kg。2)田间试验结果表明：氰霜唑在苦瓜中消解符合一级反应动力学方程,半衰期为3.0～4.9 d,属易降解农药；使用100 g/L氰霜唑悬浮剂,施药剂量105-157.5 g a.i./hm2,分别施药2、3次,苦瓜中氰霜唑总的最终残留量最大值为6.54 mg/kg。3)膳食摄入风险评估结果表明：氰霜唑的风险概率为每人每日摄入总量的1.0%。[结论]在苦瓜生长期间按照推荐剂量合理使用氰霜唑对消费者的膳食健康风险极低,对消费者健康是安全的。     

Steam radish sprout (Raphanus sativus L.): active substances,antioxidant activities and non-targeted metabolomics analysis

Various active substances, antioxidant activities and primary differential metabolite profiles were investigated to assess the nutritional quality of radish sprouts steamed for 0–300 s. The levels of flavonoids, total phenolic content and isothiocyanate in radish sprouts decreased with increased steaming time, and DPPH radical scavenging activity levels and superoxide dismutase levels also exhibited a downward trend (P < 0.05). By contrast, total phenolic acid, ferric-reducing antioxidant power and reducing power levels exhibited significant increases in the early short-term steaming process (P < 0.05). Steaming for 30 s promoted the accumulation of vitamin B₂, whereas, steaming for 300 s promoted the accumulation of phospholipids and myristic acid to a greater extent than steaming for 30 s. Overall, there was a partial non-decreased nutritional indicator in short-term steaming of radish sprouts, and a longer steaming time positively affected the differential metabolites.     

Milk metabolome reveals variations on enteric methane emissions from dairy cows fed a specific inhibitor of the methanogenesis pathway

Metabolome profiling in biological fluids is an interesting approach for exploring markers of methane emissions in ruminants. In this study, a multiplatform metabolomics approach was used for investigating changes in milk metabolic profiles related to methanogenesis in dairy cows. For this purpose, 25 primiparous Holstein cows at similar lactation stage were fed the same diet supplemented with (treated, n = 12) or without (control, n = 13) a specific antimethanogenic additive that reduced enteric methane production by 23% with no changes in intake, milk production, and health status. The study lasted 6 wk, with sampling and measures performed in wk 5 and 6. Milk samples were analyzed using 4 complementary analytical methods, including 2 untargeted (nuclear magnetic resonance and liquid chromatography coupled to a quadrupole-time-of-flight mass spectrometer) and 2 targeted (liquid chromatography-tandem mass spectrometry and gas chromatography coupled to a flame ionization detector) approaches. After filtration, variable selection and normalization data from each analytical platform were then analyzed using multivariate orthogonal partial least square discriminant analysis. All 4 analytical methods were able to differentiate cows from treated and control groups. Overall, 38 discriminant metabolites were identified, which affected 10 metabolic pathways including methane metabolism. Some of these metabolites such as dimethylsulfoxide, dimethylsulfone, and citramalic acid, detected by nuclear magnetic resonance or liquid chromatography-mass spectrometry methods, originated from the rumen microbiota or had a microbial-host animal co-metabolism that could be associated with methanogenesis. Also, discriminant milk fatty acids detected by targeted gas chromatography were mostly of ruminal microbial origin. Other metabolites and metabolic pathways significantly affected were associated with AA metabolism. These findings provide new insight on the potential role of milk metabolites as indicators of enteric methane modifications in dairy cows.     

A New and Practical Synthetic Method for the Synthesis of 6-O-Methyl-scutellarein: One Metabolite of Scutellarin in Vivo

Scutellarin (1) has been used for the treatment of angina pectoris, cerebral infarction and coronary heart disease with a large market share in China. Pharmacokinetic studies on scutellarin showed that scutellarin (1) is readily converted into its metabolites in vivo. In this paper, a new and practical synthetic method for the synthesis of 6-O-methyl-scutellarein (3) (one metabolite of scutellarin in vivo) is reported. The benzyl bromide was firstly used to selectively replace the acetyl group at C-7 in 7, and was then used to protect the hydroxy groups at C-4'' in 10, 6-O-methyl-scutellarein (3) is obtained in high yield through these methods.     

Metabonomics analysis of nonvolatile small molecules of beers during forced ageing

Three kinds of beers with different degrees of ageing were used to examine their nonvolatile small molecules by a metabonomics approach based on ultra performance liquid chromatography coupled with quadrupole time‐of‐flight tandem mass spectrometry (UPLC‐Q‐ToF‐MS/MS). Results showed that a total of 2114 compounds were detected in forced‐aged beers with the positive mode of UPLC‐Q‐ToF‐MS/MS. There was a clear separation among three groups of beers with different degrees of ageing in principal component analysis (PCA) model. Sixteen potential metabolite markers related to beer oxidative stability were identified by orthogonal partial least‐squares discriminate analysis (OPLS‐DA). Results from the changes in the spectrum also indicated that some new compounds formed in beer during forced ageing. Thus, the proposed metabonomics approach is a powerful tool to give a fully understanding the ageing process of beer based on the identification of the markers related to oxidative stability.     

吸水链霉菌A03抗菌活性产物发酵条件的优化

[目的]吸水链霉菌A03是自主分离的高效拮抗菌株,其活性代谢产物对多种植物病原真菌具有强烈抑制作用;为了建立其高产发酵工艺,并为过程放大提供技术参数,利用正交设计和单因素试验对其发酵培养基和培养条件进行优化。[结果]获得的最适发酵培养基配方(g/L):葡萄糖5,可溶性淀粉5,黄豆粉10,磷酸二氢钾0.5,碳酸钙1,七水硫酸镁0.1;最优培养条件:液体种子菌龄40 h,500 mL摇瓶装液量80 mL,接种量为体积分数12%,摇床转速180 r/min,培养温度28℃,发酵周期216 h;发酵液抑菌直径达3.04 cm。[结论]成功建立了该菌株的小试发酵工艺,发酵水平提高了52.0%,发酵周期缩短30.77%。研究结果为其发酵代谢调控和过程放大提供了基本参数。     

海洋来源真菌Fusarium sp.次级代谢产物的研究(Ⅱ)

采用多种色谱技术对海洋来源真菌Fusarium sp.的发酵物进行化学成分分离,通过波谱及理化性质分析确定了化合物的结构.从菌丝体的丙酮提取物中共分离得到10个化合物,分别为1,3,5-三羟基-7-甲基蒽醌(1)、6-羟基芦荟大黄素(2)、3-O-βD-吡喃葡萄糖基-豆甾-5,24(28)Z-二烯(3)、尿嘧啶核苷(4...