Overproduction, purification, and characterization of nanosized polyphosphate bodies from Synechococcus sp. PCC 7002

Microbial Cell Factories
Fengzheng GaoGuangxin Feng

Abstract

Inorganic polyphosphate bodies (PPB) have recently been linked to a variety of functions in mammalian cells. To improve the yield of PPB from Synechococcus sp. PCC 7002 and characterize its form, in this study, a recombinant plasmid containing a polyphosphate kinase (ppk) gene was generated and transformed into Synechococcus sp. PCC 7002. PPB separated by Sephadex G-100 was characterized and added to polarized human intestinal epithelial (Caco-2) cells, and the absorption effect was assessed. The ppk gene was stably expressed by induction with 1 μM nickel, and the resulting PPB yield from Synechococcus sp. PCC 7002 cells increased by 89.66%. Transmission electron microscopy and dynamic light scattering analyses showed that PPB from these cells were nanosized, ranging from a few to approximately 100 nanometres in diameter. PPB can be taken up by Caco-2 cells and are mainly distributed around lipid droplets. We determined that PPB can be overproduced in Synechococcus sp. PCC 7002 and that the resulting PPB were well absorbed by Caco-2 cells. Microalgae provide a promising "cell factory" for PPB production.

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Citations

Jan 22, 2019·Food & Function·Fengzheng GaoGuangxin Feng
Jul 23, 2020·Frontiers in Plant Science·Stephen P SlocombeAlison Baker
Jul 17, 2020·Frontiers in Plant Science·Emanuel Sanz-LuqueArthur R Grossman

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Methods Mentioned

BETA
PCR
electrophoresis
dynamic light scattering
transgenic
Fluorescence
genetic modification

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