Abstract
A series of culture media for haloarchaea were evaluated to optimize the production of ultrahigh-molecular-weight (UHMW) poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by Haloferax mediterranei. Cells of H. mediterranei grew (> 1 g/L of dry cell weight) and accumulated PHBV upon flask cultivation in 10 medium types with neutral pH and NaCl concentration > 100 g/L. Molecular weight and compositional analysis revealed that the number-average molecular weight (Mn) of PHBV produced with six selected types of media ranged from 0.8 to 3.5 × 106 g/mol and the 3-hydroxyvalerate (3HV) composition ranged from 8 to 36 mol%. Cultivation in two NBRC media, 1214 and 1380, resulted in the production of PHBV with an Mn of more than 3.0 × 106 g/mol and a weight-average molecular weight of more than 5.0 × 106 g/mol, indicating the production of UHMW-PHBV. These culture media contained small amount of complex nutrients like yeast extract and casamino acids, suggesting that H. mediterranei likely produced UHMW-PHBV on poor nutrient condition. Haloferax mediterranei grown in NBRC medium 1380 produced PHBV with the highest 3HV composition. A solvent-cast film of UHMW-PHBV with 26.4 mol% 3HV produced from 1-L flask cultivation with NBRC medium 1380 was found to be flexible and semi-transparent. Thermal analysis of the UHMW-PHBV cast film revealed melting and glass-transition temperatures of 90.5 °C and − 2.7 °C, respectively.
Key points
• Haloarchaeal culture media were evaluated to produce UHMW-PHBV by H. mediterranei.
• UHMW-PHBV with varied molecular weight was produced dependent on culture media.
• Semi-transparent film could be made from UHMW-PHBV with 26.4 mol% 3HV.
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The authors thank Ms. Hiroko Shinozaki for technical assistance.
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SS conceived and designed research, conducted experiments, and drafted the manuscript. KI conducted experiments and drafted the manuscript. KU discussed the results and drafted the manuscript. KW, AS, TF, KO, and TM discussed the results. All authors read and approved the manuscript.
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Sato, S., Ino, K., Ushimaru, K. et al. Evaluating haloarchaeal culture media for ultrahigh-molecular-weight polyhydroxyalkanoate biosynthesis by Haloferax mediterranei. Appl Microbiol Biotechnol 105, 6679–6689 (2021). https://doi.org/10.1007/s00253-021-11508-3
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DOI: https://doi.org/10.1007/s00253-021-11508-3