Raman-deuterium isotope probing to study metabolic activities of single bacterial cells in human intestinal microbiota

Wang, Yi1,2,3; Xu, Jiabao3; Kong, Lingchao1; Liu, Tang2; Yi, Lingbo4; Wang, Hongjuan4; Huang, Wei E.3; Zheng, Chunmiao2

2020-03

10.1111/1751-7915.13519

MICROBIAL BIOTECHNOLOGY   影响因子和分区

1751-7915

Human intestinal microbiota is important to host health and is associated with various diseases. It is a challenge to identify the functions and metabolic activity of microorganisms at the single-cell level in gut microbial community. In this study, we applied Raman microspectroscopy and deuterium isotope probing (Raman-DIP) to quantitatively measure the metabolic activities of intestinal bacteria from two individuals and analysed lipids and phenylalanine metabolic pathways of functional microorganisms in situ. After anaerobically incubating the human faeces with heavy water (D2O), D2O with specific substrates (glucose, tyrosine, tryptophan and oleic acid) and deuterated glucose, the C-D band in single-cell Raman spectra appeared in some bacteria in faeces, due to the Raman shift from the C-H band. Such Raman shift was used to indicate the general metabolic activity and the activities in response to the specific substrates. In the two individuals' intestinal microbiota, the structures of the microbial communities were different and the general metabolic activities were 76 +/- 1.0% and 30 +/- 2.0%. We found that glucose, but not tyrosine, tryptophan and oleic acid, significantly stimulated metabolic activity of the intestinal bacteria. We also demonstrated that the bacteria within microbiota preferably used glucose to synthesize fatty acids in faeces environment, whilst they used glucose to synthesize phenylalanine in laboratory growth environment (e.g. LB medium). Our work provides a useful approach for investigating the metabolic activity in situ and revealing different pathways of human intestinal microbiota at the single-cell level.

SCI

英语

通讯

Guangdong Provincial Key Laboratory of Soil and Groundwater Pollution Control[2017B030301012]

Biotechnology & Applied Microbiology ; Microbiology

Biotechnology & Applied Microbiology ; Microbiology

WOS:000513075000026

WILEY

Web of Science

1.Harbin Inst Technol, Sch Environm, Harbin 150090, Peoples R China
2.Southern Univ Sci & Technol, Sch Environm Sci & Engn, Guangdong Prov Key Lab Soil & Groundwater Pollut, Shenzhen 518055, Peoples R China
3.Univ Oxford, Dept Engn Sci, Parks Rd, Oxford OX1 3PJ, England
4.Hlth Time Gene Inst, Shenzhen 518000, Peoples R China

Wang, Yi,Xu, Jiabao,Kong, Lingchao,et al. Raman-deuterium isotope probing to study metabolic activities of single bacterial cells in human intestinal microbiota[J]. MICROBIAL BIOTECHNOLOGY,2020,13(2):572-583.

Wang, Yi.,Xu, Jiabao.,Kong, Lingchao.,Liu, Tang.,Yi, Lingbo.,...&Zheng, Chunmiao.(2020).Raman-deuterium isotope probing to study metabolic activities of single bacterial cells in human intestinal microbiota.MICROBIAL BIOTECHNOLOGY,13(2),572-583.

Wang, Yi,et al."Raman-deuterium isotope probing to study metabolic activities of single bacterial cells in human intestinal microbiota".MICROBIAL BIOTECHNOLOGY 13.2(2020):572-583.