代谢组学在乳酸菌发酵食品研究中的应用

乳酸菌是具有调节免疫、防癌抗突变的益生菌,也是食品生产中的重要工业发酵菌群。代谢组学技术已在乳酸菌发酵食品的进程监控、产品分型、风味形成机理等研究中广泛应用。本研究概述了代谢组学在食品与营养科学研究中的分离和检测技术平台以及数据分析方法,着重阐述了代谢组学技术在乳酸菌发酵食品行业中的应用现状,包括乳制品、豆制品、腌渍食品和饮品。未来代谢组学在乳酸菌发酵食品中的研究将集中在功能性乳酸菌的代谢机理、乳酸菌代谢组学数据库建设和扩容、代谢组学与基因组学、蛋白质组学等系统生物学技术结合应用等领域,尤其是代谢组学技术引入腐乳、酸菜等中国传统乳酸菌发酵食品的产品开发和品质监控,完善行业规范并提升行业发展水平。     

基于GC-MS分析两地白色藜麦种子的代谢差异

为了探究四川盐源县和青海都兰县白色藜麦种子的代谢物质差异,对其进行非靶向代谢组学的分析。利用GC-MS技术以及多元统计分析对两地的白色藜麦种子进行比较,发现两地的藜麦种子代谢物质有差异,共检测到29个显著差异物质,主要包括15个氨基酸、6个有机酸和一些糖醇、胺类、多酚、生物碱。其中有22个代谢物质表达上调,7个表达下调,在上调表达代谢物中有12个氨基酸(甘氨酸、L-缬氨酸、谷胱甘肽、GABA等)和5个有机酸(乳清酸、肉桂酸、2-氧戊二酸等)。通过MetaboAnalyst进行通路分析,发现有24条代谢通路与上调表达代谢物相关,其中有7条通路影响较显著,分别为谷胱甘肽代谢,氮代谢,丙氨酸、天冬氨酸和谷氨酸代谢,氨酰基-tRNA生物合成,苯基丙氨酸代谢,硫代谢,甘氨酸、丝氨酸和苏氨酸代谢。谷胱甘肽代谢途径的影响最为显著,而此途径与植物的抗逆性密切相关,其中检测到的代谢物主要有甘氨酸和谷胱甘肽在四川盐源的藜麦中有更高的表达水平,表明四川盐源的白色藜麦较青海都兰的白色藜麦有更强的抗氧化能力,使得植物有较好的抗逆性能在逆境中生存,这为指导两地白色藜麦的栽培育种和抗逆性机理的研究提供了理论依据。     

基于 UPLC-MS/MS 技术的线纹香茶菜 白花型与紫花型种质代谢物的比较分析酶基因转录调控研究进展

【目的】溪黄草是一种我国民间习用传统中草药,其基源植物有 3~4 种。现有的鉴别和考证研究认 为线纹香茶菜是“溪黄草”的正品之一。广泛栽培的线纹香茶菜多为紫花型,利用代谢组学技术对获得的白花型种 质与常规紫花型种质的代谢产物开展比较研究,可为正品“溪黄草”种质创制、新品种选育提供依据。【方法】采 用 UPLC-MS/MS 技术,以广泛栽培的紫花型线纹香茶菜种质为对照,比较白花型与紫花型线纹香茶菜间的差异代 谢物,并进行差异代谢物富集通路分析。【结果】从两份种质检测到的 868 种代谢物中筛选到 125 种差异代谢物, 其中,有 67 种在白花型种质中显著上调、58 种表现下调。两种质的差异代谢物主要包括黄酮类、有机酸、氨基酸 及其衍生物、酚酸类、生物碱、游离脂肪酸等,其中黄酮类化合物占比最高、为 45.6%。黄酮类、黄酮糖苷和异黄 酮三类差异代谢物在白花型种质中上调数量高于紫花型种质;而黄酮醇类化合物是两种质差异代谢物最多的类别, 且多数在紫花型种质中呈显著上调。进一步通路富集分析发现,125 种差异代谢物共注释到 47 条代谢通路,其中, 显著富集的有类黄酮（Flavonoid）、异黄酮（Isoflavonoid）、花青素（Anthocyanin）、黄酮（Flavone）、黄酮醇（Flavonol）、 芪类化合物（Stilbenoid）、二芳基庚烷（diarylheptanoid）和姜酚（Gingerol）生物合成及嘌呤（Purine）代谢等途径。 【结论】白花型线纹香茶菜种质中黄酮类、黄酮醇类化合物含量显著多于紫花型种质,将为后期线纹香茶菜种质资 源的挖掘、创制和新品种选育研究提供良好材料。     

基于UHPLC-QTOF-MS代谢组学方法筛选奶牛不同妊娠状态时的候选生物标志物

旨在筛选人工授精后第17天奶牛在不同妊娠状态时的候选生物标志物。本研究以宁夏某奶牛场体重为（550±50） kg,体况评分相近的经产（2~3胎次）健康荷斯坦奶牛为试验对象。同期发情后,在人工授精后第17天晨饲前对奶牛进行尾静脉采血,后期采用计步器和B超仪诊断出奶牛的妊娠状态。根据诊断结果将奶牛分为妊娠组（A组,n=12）和未妊娠返情组（B组,n=24）,将这两组血样进行代谢轮廓及代谢物变化分析。主成分分析（PCA）和正交偏最小二乘法-判别分析（OPLS-DA）结果显示,A、B两组血浆代谢轮廓均发生了明显变化,A、B两组之间共检测出8种ROC （Receiver operating characteristic curve）曲线下面积（area under the ROC curve,AUC）＞0.8的差异代谢物。丙氨酸-亮氨酸、甘氨酸、脯氨酸、L-正亮氨酸、DL-苯丙氨酸、肌氨酸、吡咯-2-羧酸和缬氨酸-蛋氨酸有望成为识别妊娠组（A组）和未妊娠返情组（B组）的差异代谢物。综上表明,血浆中的这8种代谢物有望成为奶牛妊娠识别阶段潜在的生物标志物,为配种早期妊娠识别提供新思路。     

Volatile Metabolite Profiling for the Discrimination of Onion Bulbs Infected by Erwinia carotovora ssp. carotovora, Fusarium oxysporum and Botrytis allii

The volatile metabolites of the headspace gas of onion bulbs inoculated with three different pathogens, Erwinia carotovora ssp. carotovora, Fusarium oxysporum and Botrytis allii, were profiled using gas chromatography/mass spectrometry. Differences in the number and amount of volatile metabolites were observed. Two hundred and fifty three volatile metabolites were detected in bulbs inoculated with three pathogens or sterile distilled water. On day three, 202 volatile metabolites were observed, compared to 166 on day six. Of the 253 compounds, however, only 59 occurred relatively consistently over replications, of which 25 compounds were specific to one or more pathogens, including 10 that were unique to a pathogen. Metabolites such as 1-Oxa-4,6-diazacyclooctane-5-thione and 4-mercapto-3-(methylthio)--(thio-lactone)-crotonic acid were exclusive to onions inoculated with F. oxysporum. Acetone, acetic acid-hydrazide, propylcarbamate, 1-bromo-1-propene, thiirane, 1-(methylthio)-E-1-propene and 1-ethenyl-4-ethyl-benzene were specific to B. allii. 3-bromo-furan was specific to E. carotovora ssp. carotovora. Sterile water-inoculated bulbs produced 3,3-dioxy-1,2-propanediol-tetranitrate. Highest amount of sulfurs was found in pathogen-inoculated, while highest amounts of terpenes, aromatics and aliphatics were found in sterile distilled water-inoculated bulbs. The possible use of these differences in the volatile metabolites for detecting and discriminating diseases of onion in storage is discussed.     

Biochemical and Metabolomics on Rice Cultivars

Plant metabolites are important for both plant life and human nutrition. However, the genetic control of plant metabolome remains largely unknown. Here, we performed a genetic analysis of the different rice metabolome and isozymes which are highly versatile and non-destructive as bio-markers. Five isozymes peroxidase, catalase, malate dehydrogenase, alcohol dehydrogenase(ADH), polyphenol oxidase were studied to characterize the thirty rice cultivars and two hybrids KRH-2 and KRH-4 along with their parental lines. Among the zymograms, ADH was found to be useful for the detection of cultivars, like CTH1, IR64, IR30864, with an Rm value of 0.549. The metabolomics of rice cultivars by using gas chromatography coupled with mass spectrometry instrument with selected reaction monitoring mode software identified the 66 metabolites in the rice samples, including amino, organic, fatty acids, alcohols and sugars(mono-/dis-accharides). All metabolites investigated varied significantly among rice samples. Jaya had the higher number of metabolites(15) with a peak for each metabolite, followed by Jyothi(13). This study demonstrated a powerful tool and provided a high-quality data for understanding the plant metabolome and isozymes, which may help bridge the gap between the genome and phenome.     

Effects of high ambient temperature on plasma metabolomic profiles in chicks

Exposure to high ambient temperature is detrimental to the poultry industry. To understand the influence from a metabolic perspective, we investigated the effects of exposure to high ambient temperature on plasma low‐molecular‐weight metabolite levels in chicks using gas chromatography/mass spectrometry‐based non‐targeted metabolomic analysis. Heat exposure for 4 days suppressed growth and food intake. Of the 92 metabolites identified, the levels of 29 decreased, whereas the levels of nine increased. We performed an enrichment analysis on the identified metabolites and found 35 candidates for metabolic processes affected by heat exposure. Among them, the sulfur amino acid metabolic pathway was clearly detected and the levels of the following related metabolites were decreased: cystathionine, cysteine, cystine, homocysteine and hypotaurine. Changes in the kynurenine pathway in tryptophan metabolism, which is linked to the immune system and oxidative stress, were also observed: kynurenine and quinolinic acid levels increased, whereas nicotinamide levels decreased. These results suggest the possible involvement of various metabolic processes in heat‐exposed chicks. Some of these metabolites would be important to understand the mechanism of biological responses to high ambient temperature in chicks.     

关中奶山羊睾丸不同发育阶段代谢物的差异分析

【目的】 试验旨在从代谢组学角度探究关中奶山羊睾丸发育代谢机制。【方法】 选择1月龄断奶雄性关中奶山羊和24月龄体成熟的雄性关中奶山羊各6只,采集睾丸组织,使用液相色谱-质谱（LC-MS）技术对睾丸组织进行化学检测,通过多维和单维分析相结合的方式对两组不同代谢物进行化学检测、对比与鉴别,通过富集差异代谢物分析筛选关中奶山羊发育代谢中的潜在关键通路。【结果】 在1和24月龄关中奶山羊睾丸组织中共筛选出334个差异代谢物,其中显著上调137个,显著下调197个。对前36个差异代谢物进行比较鉴定发现,分别为脂质和类脂质分子、有机酸及其衍生物、未分类化合物和苯丙烷和聚酮化合物四大类。经相关性分析发现,在1和24月龄关中奶山羊睾丸发育过程中,脂质和类脂质分子与甘油磷脂呈最大正相关,羧基及其衍生物与甘油磷脂呈最大负相关。对不同的代谢物进行富集分析,筛选出7条潜在的重要代谢通路,分别为肿瘤中的胆碱代谢、精氨酸和脯氨酸代谢、肿瘤中的中心碳代谢、铁死亡、谷胱甘肽代谢、氨酰-tRNA生物组成以及牛磺酸和亚牛磺酸代谢。【结论】 本试验从代谢组学角度揭示了1和24月龄关中奶山羊睾丸的差异代谢物及代谢机制,为今后哺乳动物睾丸的发育研究奠定了基础。     

苦参碱对昆明小鼠粪便及血浆代谢物的影响

基于非靶向代谢组学技术,分析腹腔注射苦参碱前后昆明小鼠粪便和血浆代谢物的差异,并通过与此前16S rDNA测序结果联合分析探究苦参碱发挥药理作用的可能机理。将20只昆明小鼠随机分为2组,分别是苦参碱处理组(MT)和生理盐水处理组(NC)。苦参碱处理组每天腹腔注射40 mg·kg^-1的苦参碱,连续给药5 d,每天给药2次。生理盐水处理组按照同样方式和体积腹腔注射生理盐水。给药第6天,分别收集各组小鼠的粪便和血浆,进行非靶向代谢组学检测,并与苦参碱处理组肠道菌群的16S rDNA测序结果进行联合分析。苦参碱处理组的粪便及血浆中均存在显著差异代谢物,粪便中共鉴定出97种,血浆中有44种。聚类分析显示,粪便中苦参碱组有35种代谢物上调,104种代谢物下调;血浆中苦参碱组有20种代谢物上调,35种代谢物下调。KEGG通路分析显示粪便及血浆差异代谢物被映射到Protein digestion and absorption等代谢途径。与16S rDNA测序分析得到的显著差异菌种嗜酸乳杆菌关联分析,结果表明粪便及血浆代谢物与嗜酸乳杆菌存在相关性。苦参碱可调节昆明小鼠的体内代谢,在粪便和血浆中均存在显著差异代谢物。这些差异代谢物及与菌群之间的互作可能是其发挥药理作用的关键。