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New Recombinant Escherichia coli Strain Tailored for the Production of Poly(3-Hydroxybutyrate) from Agroindustrial By-Products

机译:专为从农用工业副产品生产聚3-羟基丁酸酯而设计的新型重组大肠杆菌菌株

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摘要

A recombinant E. coli strain (K24K) was constructed and evaluated for poly(3-hydroxybutyrate) (PHB) production from whey and corn steep liquor as main carbon and nitrogen sources. This strain bears the pha biosynthetic genes from Azotobacter sp. strain FA8 expressed from a T5 promoter under the control of the lactose operator. K24K does not produce the lactose repressor, ensuring constitutive expression of genes involved in lactose transport and utilization. PHB was efficiently produced by the recombinant strain grown aerobically in fed-batch cultures in a laboratory scale bioreactor on a semisynthetic medium supplemented with the agroindustrial by-products. After 24 h, cells accumulated PHB to 72.9% of their cell dry weight, reaching a volumetric productivity of 2.13 g PHB per liter per hour. Physical analysis of PHB recovered from the recombinants showed that its molecular weight was similar to that of PHB produced by Azotobacter sp. strain FA8 and higher than that of the polymer from Cupriavidus necator and that its glass transition temperature was approximately 20°C higher than those of PHBs from the natural producer strains.
机译:构建了重组大肠杆菌菌株(K24K),并评估了从乳清和玉米浆作为主要碳源和氮源的聚(3-羟基丁酸酯)(PHB)的生产。该菌株带有来自Azotobacter sp。的pha生物合成基因。由乳糖操纵者控制下的T5启动子表达的FA8菌株。 K24K不产生乳糖阻遏物,从而确保参与乳糖运输和利用的基因的组成型表达。通过在实验室规模的生物反应器中在补料分批培养中需氧生长的重组菌株在补充了农用工业副产物的半合成培养基上有效地产生有氧菌种,可以有效生产PHB。 24小时后,细胞积累的PHB达到其细胞干重的72.9%,达到了每升每小时2.13 g PHB的体积生产率。从重组物中回收的PHB的物理分析表明,其分子量类似于Azotobacter sp。生产的PHB。菌株FA8,比来自Cupriavidus necator的聚合物要高,其玻璃化转变温度比天然生产菌株的PHB高约20°C。

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