Transcriptome analysis of polysaccharide-based microbial flocculant MBFA9 biosynthesis regulated by nitrogen source

Microbial flocculant (MBF), an environmentally friendly water treatment agent, can be widely used in various water treatments. However, its use is limited by low yield and high cost. This problem can be solved by clarifying its biosynthesis mechanism and regulating it. Paenibacillus shenyangensis A9...

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Bibliographic Details
Published in:	Scientific reports 2020-02, Vol.10 (1), p.2918, Article 2918
Main Authors:	Fu, Lili, Jiang, Binhui, Wei, Jianwei, Liu, Jinliang, Hu, Xiaomin, Zhang, Li
Format:	Article
Language:	eng
Subjects:	Bacteria Biosynthesis Biosynthetic Pathways - drug effects Biosynthetic Pathways - genetics Carbon - pharmacology Fermentation Flocculation Gene expression Gene Expression Profiling Gene Expression Regulation, Bacterial - drug effects Gene Ontology Gene regulation Genes, Bacterial Genomes Metabolic pathways Metabolism Next-generation sequencing Nitrogen Nitrogen - pharmacology Paenibacillus Paenibacillus - chemistry Paenibacillus - drug effects Paenibacillus - genetics Paenibacillus - growth & development Polysaccharides Polysaccharides - pharmacology Ribonucleic acid RNA Water treatment
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Summary:	Microbial flocculant (MBF), an environmentally friendly water treatment agent, can be widely used in various water treatments. However, its use is limited by low yield and high cost. This problem can be solved by clarifying its biosynthesis mechanism and regulating it. Paenibacillus shenyangensis A9, a flocculant-producing bacterium, was used to produce polysaccharide-type MBFA9 by regulating the nitrogen source (nitrogen adequacy/nitrogen deficiency). In this study, RNA-Seq high-throughput sequencing technology and bioinformatic approaches were used to investigate the fermentation and biosynthesis of polysaccharide-type MBFA9 by regulating the nitrogen source (high nitrogen/low nitrogen) in the flocculant-producing bacteria Paenibacillus shenyangensis A9. Differentially expressed genes, functional clustering, and functional annotation of key genes were assessed. Then the MBFA9 biosynthesis and metabolic pathway were reconstructed. Our results showed that when cultured under different nitrogen conditions, bacterial strain A9 had a greater ability to synthesize polysaccharide-type MBFA9 under low nitrogen compared to high nitrogen conditions, with the yield of MBFA9 reaching 4.2 g/L at 36 h of cultivation. The quality of transcriptome sequencing data was reliable, with a matching rate of 85.38% and 85.48% when L36/H36 was mapped to the reference genome. The total expressed genes detected were 4719 and 4730, with 265 differentially expressed genes. The differentially expressed genes were classified into 3 categories: molecular function (MF), cell component (CC), and biological process (BP), and can be further divided into 22 subcategories. There were 192 upregulated genes and 73 downregulated genes, with upregulation being predominant under low nitrogen. UDP-Gal, UDP-Glc, UDP-GlcA, and UDP-GlcNAc, which are in the polysaccharide metabolic pathway, could all be used as precursors for MBFA9 biosynthesis, and murA, wecB, pgm, galU/galF, fcl, gmd, and glgC were the main functional genes capable of affecting the growth of bacteria and the biosynthesis of MBF. Results from this study provide evidence that high-level expression of key genes in MBFA9 biosynthesis, regulation, and control can achieve MBFA9 directional synthesis for large-scale applications.
ISSN:	2045-2322 2045-2322