Mezzina Mariela P, Álvarez Daniela S, Egoburo Diego E, Díaz Peña Rocío, Nikel Pablo I, Pettinari M Julia
Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Departamento de Química Biológica, Buenos Aires, Argentina.
Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales (CONICET-Universidad de Buenos Aires), Buenos Aires, Argentina.
Appl Environ Microbiol. 2017 Jun 30;83(14). doi: 10.1128/AEM.00662-17. Print 2017 Jul 15.
The microbial production of biofuels and other added-value chemicals is often limited by the intrinsic toxicity of these compounds. The phasin PhaP from the soil bacterium sp. strain FA8 is a polyhydroxyalkanoate granule-associated protein that protects recombinant against several kinds of stress. PhaP enhances growth and poly(3-hydroxybutyrate) synthesis in polymer-producing recombinant strains and reduces the formation of inclusion bodies during overproduction of heterologous proteins. In this work, the heterologous expression of this phasin in was used as a strategy to increase tolerance to several biotechnologically relevant chemicals. PhaP was observed to enhance bacterial fitness in the presence of biofuels, such as ethanol and butanol, and other chemicals, such as 1,3-propanediol. The effect of PhaP was also studied in a mutant strain, in which both GroELS and PhaP were observed to exert a beneficial effect that varied depending on the chemical tested. Lastly, the potential of PhaP and GroEL to enhance the accumulation of ethanol or 1,3-propanediol was analyzed in recombinant Strains that overexpressed either or had increased growth, reflected in a higher final biomass and product titer than the control strain. Taken together, these results add a novel application to the already multifaceted phasin protein group, suggesting that expression of these proteins or other chaperones can be used to improve the production of biofuels and other chemicals. This work has both basic and applied aspects. Our results demonstrate that a phasin with chaperone-like properties can increase bacterial tolerance to several biochemicals, providing further evidence of the diverse properties of these proteins. Additionally, both the PhaP phasin and the well-known chaperone GroEL were used to increase the biosynthesis of the biotechnologically relevant compounds ethanol and 1,3-propanediol in recombinant These findings open the road for the use of these proteins for the manipulation of bacterial strains to optimize the synthesis of diverse bioproducts from renewable carbon sources.
生物燃料和其他增值化学品的微生物生产常常受到这些化合物内在毒性的限制。来自土壤细菌sp. 菌株FA8的相蛋白PhaP是一种与聚羟基脂肪酸酯颗粒相关的蛋白质,可保护重组体免受多种应激。PhaP可促进聚合物生产重组菌株的生长和聚(3-羟基丁酸酯)合成,并减少异源蛋白过量生产期间包涵体的形成。在这项工作中,该相蛋白在中的异源表达被用作提高对几种生物技术相关化学品耐受性的策略。观察到PhaP在生物燃料(如乙醇和丁醇)以及其他化学品(如1,3-丙二醇)存在的情况下可增强细菌适应性。还在一个突变菌株中研究了PhaP的作用,在该菌株中观察到GroELS和PhaP均发挥了有益作用,其作用因所测试的化学品而异。最后,在重组菌株中分析了PhaP和GroEL增强乙醇或1,3-丙二醇积累的潜力。过表达或的菌株生长增加,表现为最终生物量和产物滴度高于对照菌株。综上所述,这些结果为已经具有多方面功能的相蛋白家族增加了一种新的应用,表明这些蛋白或其他伴侣蛋白的表达可用于改善生物燃料和其他化学品的生产。这项工作具有基础和应用两个方面。我们的结果表明,具有伴侣样性质的相蛋白可提高细菌对几种生化物质的耐受性,为这些蛋白质的多样性质提供了进一步证据。此外,PhaP相蛋白和著名的伴侣蛋白GroEL均被用于增加重组中生物技术相关化合物乙醇和1,3-丙二醇的生物合成。这些发现为利用这些蛋白质操纵细菌菌株以优化从可再生碳源合成多种生物产品开辟了道路。