Method of delivery--Detection and significance of metabolites in intestinal flora
Professor Jia Wei from the Translational Medicine Center of the Sixth People's Hospital affiliated to Shanghai Jiaotong University has made new progress in the detection of intestinal flora metabolites, and developed a gas chromatography-time-of-flight mass spectrometry (GC-TOF/ MS) High-throughput absolute quantitative detection of 150 important gut microbial metabolites by microbial metabolomics analysis in samples of serum, urine, feces or bacteria (eg E. coli) in 15 minutes The intestinal flora metabolites were subjected to fully automated chemical derivatization and quantitative analysis. These metabolites include amino acids, fatty acids, organic acids, phenols, phenyl or benzyl derivatives, hydrazine, etc., and are involved in a number of important metabolic pathways involved in the metabolism of the gut flora. The research team also constructed a database of methyl chloroformate and ethyl chloroformate derivatives of the above metabolites. All metabolites contained in the library have complete information on mass spectrometry fragments and retention indices of the above two derivatives. The analysis of the mass spectrometry fragmentation patterns of these two derivatives not only greatly improved the accuracy of metabolite identification, but also provided a reference for speculating the structure of unknown compounds. This method has been published online in the international analytical chemistry mainstream journal Analytical Chemistry. The establishment of this automated analytical platform is of great significance for the study of the relationship between human intestinal microecology and health and disease.
Why do you need a simple, fast, and accurate method for detecting intestinal flora metabolites?
In the human gastrointestinal tract, a complex and large number of microbial populations have been born. Through long-term co-evolution with humans (hosts), they have become important "functional organs" that can not be ignored in human health. More and more research evidence indicates that intestinal flora imbalance is closely related to the occurrence of various diseases such as obesity, diabetes, nonalcoholic fatty liver, inflammatory bowel disease and digestive tract cancer. Therefore, exploring the relationship between changes in intestinal flora metabolites and host disease is of great significance for the prevention and treatment of diseases.
The emerging field of microbiology has received increasing attention in recent years, but the current characterization of bacteria in this field is basically based on sequencing technologies such as 16S rRNA sequencing or NextGen macro sequencing. Sequencing can tell us only the type information of intestinal bacteria, the classification and abundance values ​​from the door to the genus to the species (sometimes even to the strain), but the functional information of the bacteria, especially the composition of the bacterial community. We do not know the overall metabolic function of the ecology and how it interacts with our human body (host). We can't judge the difference between the two intestines with different composition of the gut flora. What are the structural differences? It is difficult to predict what kind of function these two groups will have in the metabolism and physiology of the two people on the same diet. Sexual change. The most straightforward method of characterizing metabolic function is to measure the metabolome. The data obtained are the integrated functions of various bacteria and the final result of working with the host!
Microbial metabolomics uses metabolomics analysis techniques and data processing methods to understand the physiological state of microorganisms through qualitative and quantitative analysis of microbial metabolites, and to explore the potential interaction mechanisms between intestinal microbes and hosts and intestinal tracts. The effect of bacterial metabolism on host health and disease.
What are our intestinal bacterial metabolites?
The main substrate for intestinal flora metabolism is carbohydrates, proteins and peptides in food that cannot be digested and absorbed by the small intestine. Dietary fiber in food can be fermented and utilized by bacteria to produce monosaccharide and oligosaccharide molecules, or organic acids such as ethanol, lactic acid, succinic acid, etc., and further form short-chain fatty acids such as acetic acid, propionic acid, butyric acid. These short-chain fatty acids are not only a source of energy for the host and microorganisms, but also can be metabolized by a variety of peripheral tissues. Proteins in food can be broken down by intestinal bacteria to produce peptides and amino acids. Among them, branched chain amino acids such as proline, leucine and isoleucine can be further metabolized by bacteria to form branched fatty acids, and bacterial metabolites of aromatic amino acids such as phenylalanine, tryptophan and tyrosine are Phenols and anthraquinones. In addition, the synthesis of various vitamins such as vitamin K and some B vitamins in the human body is also associated with intestinal flora metabolism.
Intestinal flora can not only metabolize substances that the host itself cannot metabolize, but also participate in the metabolism of the host and co-metabolize with the host to form a series of intestinal flora-host co-metabolites, such as cholesterol and bile acid metabolism, hormones. Metabolism and the like are all done together by the intestinal flora and the host. So far, studies have confirmed that metabolites related to intestinal flora metabolism and its co-metabolism with the host mainly include short-chain fatty acids, bile acids (especially secondary bile acids), and choline metabolites (such as trimethylamine-N- Oxide), phenols, phenyl (or benzyl) derivatives, terpenoids, polyamines, lipids (such as conjugated fatty acids), vitamins, hormones, and the like.
Accurate, sensitive, fast, and high-throughput microbial metabolomics is a core tool for precision medicine in the future!
Due to the complexity and diversity of the structure and properties of the intestinal flora-host co-metabolites, it is technically challenging to simultaneously measure numerous co-metabolites with as few detection platforms as possible. In view of the fact that most of these metabolites are highly polar and difficult to volatilize, we have developed this proprietary fully automated derivatization method to selectively derivate the microbial-host co-metabolites effectively, effectively eliminating exogenous drugs and The interference of other sugar substances, the method is easy to operate, and is beneficial to the processing of large quantities of samples. This method based on GC-TOF/MS platform for high-throughput absolute quantitative detection of intestinal flora-host co-metabolites in biological samples has the characteristics of high practicability, short analysis time and high detection flux, which can be used for microbial-host Metabolites were subjected to rapid quasi-deterministic quantitative analysis.
Combine metabolomics with microbiome and molecular biology methods to find co-variation of metabolic phenotypic changes and bacterial structure changes, identify and identify functional bacteria that have significant effects on host physiological metabolism, and establish them at the system biology level. The association model between host metabolism and intestinal flora can objectively detect the metabolic components and concentration changes of intestinal microbes and display the metabolic status of intestinal bacteria, thus allowing us to study the relationship between intestinal flora and host more deeply. A complex metabolic system that understands how the intestinal microbiota affects the metabolic state of the host through its own metabolism and co-metabolism with the host.
The construction of microbial metabolomics analysis platform provides a basis for studying the correlation between metabolic phenotypic dynamics and microbial composition changes. By analyzing the interaction of the flora and the host at the metabolic level, observing the changes of key metabolites, and facilitating researchers to understand more deeply the complex mechanism of intestinal flora, this system biology research model can accelerate non- The development of invasive disease diagnostic tools, promote individualized medical processes, improve the effectiveness of individualized therapeutics, and accelerate the precise medical process of intestinal flora research to clinical transformation.
Related literature:
- Zhao, LJ, Ni, Y., Su, MM, Li, HS, Dong, FC, Chen, WL, Wei, RM,, Zhang, LL, Guiraud, SP, Martin, FJ, Rajani, C., Xie, GX , Jia, W.* High throughput and quantitative measurement of microbial metabolome by gas chromatography/mass spectrometry using automated alkyl chloroformate derivatization. Analytical Chemistry . doi: 10.1021/acs.analchem.7b00660.
- Nicholson, JK, Holmes, E., Kinross, J., Burcelin, R., Gibson, G., Jia, W., Pettersson, S. Host-gut microbiota metabolic interactions. Science, 336(6086): 1262-7 , 2012.
- Zheng, XJ, Xie, GX,, Zhao, AH, Zhao, LJ, Yao, C., Chiu, NH, Zhou, ZX, Bao, YQ, Jia, WP, Nicholson, JK, Jia, W*. The footprints of Gut microbial-mammalian co-metabolism. Journal of Proteome Research , 10(12): 5512-22, 2011.
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