Biosynthesis of Polyhydroxyalkanoates from Defatted Chlorella Biomass as an Inexpensive Substrate

Abstract

Microalgae biomass has been recently used as an inexpensive substrate for the industrial production of polyhydroxyalkanoates (PHAs). In this work, a dilute acid pretreatment using 0.3 N of hydrochloric acid (HCl) was performed to extract reducing sugars from 10% (w/v) of defatted Chlorella biomass (DCB). The resulting HCl DCB hydrolysate was used as a renewable substrate to assess the ability of three bacterial strains, namely Bacillus megaterium ALA2, Cupriavidus necator KCTC 2649, and Haloferax mediterranei DSM 1411, to produce PHA in shake flasks. The results show that under 20 g/L of DCB hydrolysate derived sugar supplementation, the cultivated strains successfully accumulated PHA up to 29.7–75.4% of their dry cell weight (DCW). Among the cultivated strains, C. necator KCTC 2649 exhibited the highest PHA production (7.51 ± 0.20 g/L, 75.4% of DCW) followed by H. mediterranei DSM 1411 and B. megaterium ALA2, for which a PHA content of 3.79 ± 0.03 g/L (55.5% of DCW) and 0.84 ± 0.06 g/L (29.7% of DCW) was recorded, respectively. Along with PHA, a maximum carotenoid content of 1.80 ± 0.16 mg/L was produced by H. mediterranei DSM 1411 at 120 h of cultivation in shake flasks. PHA and carotenoid production increased by 1.45- and 1.37-fold, respectively, when HCl DCB hydrolysate biotransformation was upscaled to a 1 L of working volume fermenter. Based on FTIR and 1H NMR analysis, PHA polymers accumulated by B. megaterium ALA2 and C. necator KCTC 2649 were identified as homopolymers of poly(3-hydroxybutyrate). However, a copolymer of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with a 3-hydroxyvalerate fraction of 10.5 mol% was accumulated by H. mediterranei DSM 1411.