Citation: ZENG Ting-Lan, YE Yang-Fang, MU Chang-Kao, WANG Kai, LI Rong-Hua, WANG Chun-Lin. Gut Microbiota and Metabolic Phenotype of Portunus trituberculatus[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(12): 1867-1873. doi: 10.11895/j.issn.0253-3820.160432 shu

Gut Microbiota and Metabolic Phenotype of Portunus trituberculatus

1.
Key Laboratory of Applied Marine Biotechnology(Ningbo University), Ministry of Education, Ningbo 315211, China;
2.
State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan 430071, China

Received Date: 6 June 2016
Revised Date: 30 August 2016

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 41673076, 41476124), the sci. & Tech. Project for Common Wealth of Ningbo (No. 2016C10062), and the K.C, Wang Magna Found of Ningbo University.

16S rRNA gene high-throughput sequencing and nuclear magnetic resonance (NMR)-based metabolomics techniques were established to understand the composition of gut microbiota and metabolites of Portunus trituberculatus. Swimming crab gut samples were used for the extraction of bacterial genomic DNA, followed by the amplification of hypervariable domain V3-V4 of 16 S rRNA gene. The obtained PCR products were used for analysis on an Illumina MiSeq platform. The bacterial phylotypes were then clustered and assigned. The results showed that swimming crab gut was dominated by proteobacteria, bacteroidetes, fusobacteria, tenericutes, and acidobacteria at the phylum level. At the genus level, Photobacterium, Paludibacter, and Propionigenium were enriched in it. Crab intestinal samples were extracted with CH₃OH/H₂O (2:1, V/V) solution by shaking with a tissuelyser. After the removal of methanol, the resultant supernatants were lyophilized. Each of lyophilized extract was dissolved into Na⁺/K⁺ phosphate buffer and then centrifuged at high speed for NMR analysis. A standard noesypr1D was used to acquire ¹H NMR spectra. The 90° pulse length was adjusted to approximately 10 μs. The recycle delay and mixing time were set to 2 s and 100 ms in sequence. The spectral width was set to 20 ppm. Sixty-four transients were collected into 32 k data points for each spectrum. A range of two-dimensional NMR spectra were acquired for the resonance assignment. The results showed that swimming crab gut metabolome comprised 30 metabolites including some amino acids, organic acids, and amines. In summary, this study provided a method for the systematic analysis of the compositions of gut bacterial community and metabolites of swimming crab.
- Portunus trituberculatus,
- Gut microbiota,
- High-throughput sequencing,
- Metabolic phenotype,
- Nuclear magnetic resonance
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沈阳化工大学材料科学与工程学院沈阳 110142