Abstract:

A high-throughput method for cell phenotype analysis was developed based on 96-well plate culture and gas chromatography coupled with mass spectrometry (GC-MS). Forty-eight metabolites were selected as the sole energy for wild-type and its yfcC mutation E. coli (yfcC over-expression and yfcC deletion mutants) culture. The high-throughput phenotype analysis was achieved by investigating the catabolism difference of these metabolites among the three strains by GC-MS. The results showed that there were 14 metabolites changed significantly between wild-type and yfcC over-expression mutant E. coli. The metabolism ability of yfcC over-expression mutant E. coli for glycine and citric acid was much stronger than wild-type E. coli. For other metabolites, the wild-type strain showed stronger capabilities in metabolism. Among the 16 metabolites metabolized differentially between wild-type E. coli and its yfcC deletion mutant, metabolism ability of alanine, lactose, myo-inositol and citric acid in yfcC deletion mutant was much stronger. Other metabolites displayed the opposite results. Among the results acquired, we speculated that the faster metabolism of glycine in yfcC over-expression mutant E. coli might be caused by its promoted glyoxylate shunt. This high-throughput method for cell phenotype analysis was simple and efficient. It could provide more useful information about metabolism for gene study with unknown function.

Key words: gas chromatography-mass spectrometry (GC-MS), gene function, high-throughput, phenotype