Metabolomic Analysis of Three Mollicute Species

Abstract

We present a systematic study of three bacterial species that belong to the class Mollicutes, the smallest and simplest bacteria, Spiroplasma melliferum, Mycoplasma gallisepticum, and Acholeplasma laidlawii. To understand the difference in the basic principles of metabolism regulation and adaptation to environmental conditions in the three species, we analyzed the metabolome of these bacteria. Metabolic pathways were reconstructed using the proteogenomic annotation data provided by our lab. The results of metabolome, proteome and genome profiling suggest a fundamental difference in the adaptation of the three closely related Mollicute species to stress conditions. As the transaldolase is not annotated in Mollicutes, we propose variants of the pentose phosphate pathway catalyzed by annotated enzymes for three species. For metabolite detection we employed high performance liquid chromatography coupled with mass spectrometry. We used liquid chromatography method - hydrophilic interaction chromatography with silica column - as it effectively separates highly polar cellular metabolites prior to their detection by mass spectrometer.

Reconstruction of Transcription Control Networks in Mollicutes by High-Throughput Identification of Promoters

“…Spiroplasma melliferum KC3 was cultivated in SP4 medium (liquid): tryptone 10 g/L, peptone 10 g/L, sorbitol 70 g/L, yeast extract 7 g/L, brain-heart infusion 2.5 g/L, NaCl 4.5 g/L, sucrose 1%, fructose 1%, glucose 0.8%, horse serum 10%, and pH 7.6 at 30°C in aerobic conditions (Vanyushkina et al, 2014). For the experiment, cells were cultivated until stationary phase (approximately 24 h), and the next passage was performed in 1:10 dilution.…”

“…Mollicutes show a significant reduction in metabolic pathways, which primarily concentrate around the generation of energy (Fisunov et al, 2011). Most Mollicutes import all nutrients and cofactors due to a complete loss of respective synthetic pathways (Vanyushkina et al, 2014). Recent progress of bacterial genomics indicate that a minimal set of genes can instead be viewed as a minimal set of functions in which the same function may be carried out by different sets of genes.…”

Draft Genome Sequence of “ Candidatus Hepatoplasma crinochetorum” Ps, a Bacterial Symbiont in the Hepatopancreas of the Terrestrial Isopod Porcellio scaber

“…Nucleotide synthesis and many pathways for amino acid, vitamin, and cofactor biosynthesis and cell wall synthesis are degenerated. Energy is likely produced by the introduction of various sugars into glycolysis and the arginine deiminase pathway ( 10 ). The draft genome sequence of “ Ca.…”

GC/MS-based metabolic profiling reveals important metabolic pathways in microsporidia, Nosema antheraeae

“…It becomes a major component of systems biology. Up to now, it is a powerful tool not only for metabolite identification (VANYUSHKINA & al [6]) , but also for the areas closely associated with human health and nursing, such as disease diagnosis (HUANG & al [7]), nutrition and food science, environmental science and so on. Most notably, metabolomics could not only play an important role in exploring the mechanisms of biological response (Barding & al [8]), but also provide clues to mechanistic research (CHEN & al [9]).…”