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Biotechnological synthesis of water-soluble food-grade polyphosphate with Saccharomyces cerevisiae.
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2020-03-19 , DOI: 10.1002/bit.27337
Jonas Johannes Christ 1 , Stephanie A Smith 2 , Sabine Willbold 3 , James H Morrissey 2 , Lars Mathias Blank 1
Affiliation  

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.

中文翻译:

用酿酒酵母生物技术合成水溶性食品级多磷酸盐。

无机多磷酸盐 (polyP) 是磷酸盐的聚合物。平均链长为 2-40 个 P 亚基的水溶性聚磷被广泛用作食品添加剂,目前是通过化学合成的。本研究提出了一种环境友好的高度可扩展的粉末形式水溶性食品级聚磷(称为生物聚磷)的生物合成工艺。在无磷酸盐培养基中孵育后,通常被认为是安全的野生型面包酵母(酿酒酵母)吸收磷酸盐并在细胞内聚合成 26.5% 的聚磷(作为 KPO 3,以细胞干重计)。通过冻融和温和热处理(10 分钟,70°C)裂解细胞。通过用 50 mM HCl 沉淀从可溶性细胞成分中去除蛋白质和核酸。通过分级 polyP 沉淀(组分 1 用 100 mM NaCl 和 0.15 体积乙醇沉淀,组分 2 用 1 vol 乙醇),干燥和研磨。bio-polyP 的理化性质通过酶分析、31 P 核磁共振光谱和聚丙烯酰胺凝胶电泳等进行分析。该工艺的一个设想应用是将废物流中的磷酸盐回收成高价值的生物多聚磷。
更新日期:2020-03-19
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