Rapid Quantification of Polyhydroxybutyrate Polymer from Single Bacterial Cells with Mass Spectrometry.

Abstract

Polyhydroxyalkanoate (PHA) is one of the biocompatible and biodegradable plastics that can be produced and accumulated as granules inside microorganisms. In this study, a new approach to rapidly quantify a short-chain-length PHA, polyhydroxybutyrate (PHB), produced from genetically engineered Escherichia coli containing phaCAB is presented. The mass of each bacterial cell was measured using a laser-induced radio frequency (rf) plasma charge detection quadrupole ion trap mass spectrometer (LIRFP CD QIT-MS), and then, the PHB contents were determined by calculating the change in cellular mass. The quantitative results showed that the PHB contents measured by LIRFP CD QIT-MS were consistent with those by reference analysis, gas chromatography (GC). The PHB content of each bacterial sample can be obtained within 20 min from sampling using LIRFP CD QIT-MS while GC analysis takes 2 days. In addition, LIRFP CD QIT-MS does not use any hazardous chemicals in cellular mass quantification as compared to GC. This indicates that LIRFP CD QIT-MS has potential in routine monitoring of PHB production.