Biotechnological synthesis of water‐soluble food‐grade polyphosphate with Saccharomyces cerevisiae

Abstract

Inorganic polyphosphate (polyP) is the polymer of phosphate. Water‐soluble polyPs with average chain lengths of 2–40 P‐subunits are widely used as food additives and are currently synthesized chemically. An environmentally friendly highly scalable process to biosynthesize water‐soluble food‐grade polyP in powder form (termed bio‐polyP) is presented in this study. After incubation in a phosphate‐free medium, generally regarded as safe wild‐type baker's yeast (Saccharomyces cerevisiae) took up phosphate and intracellularly polymerized it into 26.5% polyP (as KPO3, in cell dry weight). The cells were lyzed by freeze‐thawing and gentle heat treatment (10 min, 70°C). Protein and nucleic acid were removed from the soluble cell components by precipitation with 50 mM HCl. Two chain length fractions (42 and 11P‐subunits average polyP chain length, purity on a par with chemically produced polyP) were obtained by fractional polyP precipitation (Fraction 1 was precipitated with 100 mM NaCl and 0.15 vol ethanol, and Fraction 2 with 1 final vol ethanol), drying, and milling. The physicochemical properties of bio‐polyP were analyzed with an enzyme assay, 31P nuclear magnetic resonance spectroscopy, and polyacrylamide gel electrophoresis, among others. An envisaged application of the process is phosphate recycling from waste streams into high‐value bio‐polyP.

Characterization of Agrobacterium tumefaciens PPKs reveals the formation of oligophosphorylated products up to nucleoside nona-phosphates

“…In most prokaryotes, polyP is present in form of granule-like inclusions (polyP granules or voluntin granules) with diameters mostly in the range of 50 to 200 nm. In yeasts, polyP can be accumulated up to ≈ one quarter of the cellular weight in vacuole-like compartments (Christ et al 2020a). PolyP represents a reservoir of phosphorous and apparently is also involved in various forms of stress resistance such as tolerance against heavy metals, elevated temperature, or reactive oxygen species and can fulfill functions in virulence, motility, biofilm formation, and cell cycle control Nikel et al 2013;Chuang et al 2013;Gray and Jakob 2015;Racki et al 2017).…”

New developments in biological phosphorus accessibility and recovery approaches from soil and waste streams

“…The highest amount of phytase‐released P was 9.97 g P kg –1 of pressed cake. Poly‐P can also be enriched in Saccharomyces cerevisiae after a starvation procedure and extracted and purified as poly‐P chains of different lengths [101, 102]. For plants, a bioengineering approach in conventional plant breeding can be steered into a direction where plants can produce higher amounts of organic acids and enzymes to facilitate better soil P immobilization and utilization [1].…”

Sustainable Production of Reduced Phosphorus Compounds: Mechanochemical Hydride Phosphorylation Using Condensed Phosphates as a Route to Phosphite

“…In biological systems, long-chain polyphosphate (polyP) is involved in ATP synthesis catalyzed by polyP kinases, in which polyP is the source of a third PO 3 – group leading to ATP. , Inspired by the biochemical reactivity of polyP, we turned our attention to the application of bioproduced polyP as a source of PO 3 – . A recent protocol of polyP accumulation using Saccharomyces cerevisiae (baker’s yeast) followed by isolation allowed us to obtain bioproduced polyP (bio-polyP, Figure ) in multigram quantities. , The composition and structure of the isolated bio-polyP was analyzed by 31 P NMR spectroscopy in conjunction with several elemental analytical methods. Linear phosphates (with Na + , Mg 2+ , K + , and Ca 2+ as counterions) constitute 77.8 wt % of bio-polyP with an average degree of polymerization of 37, whereas residual orthophosphate and trimetaphosphate are <1.5 wt %.…”

“…A recent protocol of polyP accumulation using Saccharomyces cerevisiae (baker's yeast) followed by isolation allowed us to obtain bioproduced polyP (bio-polyP, Figure 3) in multigram quantities. 82,83 The composition and structure of the isolated bio-polyP was analyzed by 31 P NMR spectroscopy in conjunction with several elemental analytical methods. Linear phosphates (with Na + , Mg 2+ , K + , and Ca 2+ as counterions) constitute 77.8 wt % of bio-polyP with an average degree of polymerization of 37, whereas residual orthophosphate and trimetaphosphate are <1.5 wt %.…”