Chemicals Affecting Cyanobacterial Poly(3-hydroxybutyrate) Accumulation: 2-Phenylethanol Treatment Combined with Nitrogen Deprivation Synergistically Enhanced Poly(3-hydroxybutyrate) Storage in Synechocystis sp. PCC6803 and Anabaena sp. TISTR8076

Abstract

Abstract Various photoautotrophic cyanobacteria increase the accumulation of bioplastic poly(3-hydroxybutyrate) (PHB) under nitrogen deprivation (–N) for energy storage. Several metabolic engineering enhanced cyanobacterial PHB accumulation, but these strategies are not applicable in non-gene-transformable strains. Alternatively, stimulating PHB levels by chemical exposure is desirable because it might be applied to various cyanobacterial strains. However, the study of such chemicals is still limited. Here, 19 compounds previously reported to affect bacterial cellular processes were evaluated for their effect on PHB accumulation in Synechocystis sp. PCC6803, where 3-(3,4-dichlorophenyl)-1,1-dimethylurea, methyl viologen, arsenite, phenoxyethanol and 2-phenylethanol were found to increase PHB accumulation. When cultivated with optimal nitrate supply, Synechocystis contained less than 0.5% [w/w dry weight (DW)] PHB, while cultivation under –N conditions increased the PHB content to 7% (w/w DW). Interestingly, the –N cultivation combined with 2-phenylethanol exposure reduced the Synechocystis protein content by 27% (w/w DW) but significantly increased PHB levels up to 33% (w/w DW), the highest ever reported photoautotrophic cyanobacterial PHB accumulation in a wild-type strain. Results from transcriptomic and metabolomic analysis suggested that under 2-phenylethanol treatment, Synechocystis proteins were degraded to amino acids, which might be subsequently utilized as the source of carbon and energy for PHB biosynthesis. 2-Phenylethanol treatment also increased the levels of metabolites required for Synechocystis PHB synthesis (acetyl-CoA, acetoacetyl-CoA, 3-hydroxybutyryl-CoA and NADPH). Additionally, under –N, the exposure to phenoxyethanol and 2-phenylethanol increased the PHB levels of Anabaena sp. from 0.4% to 4.1% and 6.6% (w/w DW), respectively. The chemicals identified in this study might be applicable for enhancing PHB accumulation in other cyanobacteria.