Heterologous production of Escherichia coli penicillin G acylase in Pseudomonas aeruginosa

Krzeslak, Joanna; Braun, Peter; Voulhoux, Romé; Cool, Robbert H.; Quax, Wim J.

doi:10.1016/j.jbiotec.2009.05.005

Cited by 14 publications

(2 citation statements)

References 61 publications

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“…It thus seems more likely that PA does not rely fully (if at all) on the Tat pathway, but mainly uses the most common route for export in E. coli, the Sec pathway (Natale et al 2008). Interestingly, E. coli PA is also transported independently of the Tat pathway in Pseudomonas aeruginosa (Krzeslak et al 2009), which corroborates our suggestion that PA is a Sec-dependent protein. Tables Table 1 Primers used in this study…”

Section: Discussionsupporting

confidence: 91%

Mutations in the translation initiation region of the pac gene resulting in increased levels of activity of penicillin G acylase

Akkaya

Öztürk

Bolhuis

et al. 2012

World J Microbiol Biotechnol

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Penicillin G acylase (PA) is an important enzyme used in the industrial production of b-lactam antibiotics. In this study, the effects of mutations in the translation initiation region of the Escherichia coli pac gene, encoding periplasmic PA, were examined. Several mutations led to increased amounts of PA activity, including those that lengthened the spacer region between the ribosome binding site and the ATG start codon, and those with altered codons on positions +2 and +4 relative to the start codon. These results indicated that the wild-type sequence of the pac gene does not provide maximum expression levels and that the strategies applied in this study can be used to improve production of PA in E. coli. Unexpectedly, our study also suggested that translocation of PA was, in contrast to earlier reports, shown not to require the Twin-arginine translocation pathway for transport into the periplasm.

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Section: Discussionsupporting

confidence: 91%

Mutations in the translation initiation region of the pac gene resulting in increased levels of activity of penicillin G acylase

Akkaya

Öztürk

Bolhuis

et al. 2012

World J Microbiol Biotechnol

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“…Para solucionar dicho problema, fusionaron la proteína PGA con péptidos señales Sec o Tat de Pseudomonas, logrando que la proteína quimérica TatProPGA se acumulara en el periplasma. 53 Streptomyces lividans. Uno de los aspectos críticos en la producción de proteínas recombinantes es la identificación y la purificación.…”

Section: Sistema De Expresión Inducción (Intervalo) Nivel De Expresió...unclassified

Las bacterias como sistema de expresión de proteínas heterólogas terapéuticas: una revisión bibliográfica

N.²

2014

Hech Microb.

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En la actualidad la producción de proteínas terapéuticas se ha convertido en uno de los campos de mayor impacto a nivel científico y biotecnológico. La expresión de proteínas recombinantes en sistemas procariontes, particularmente en E. coli han permitido el desarrollo de una gran variedad de proteínas terapéuticas. Sin embargo, para poder producir proteínas funcionales ha sido necesario buscar intensamente un equilibrio entre la calidad y la producción; por lo que es necesario innovar nuevas estrategias que permitan superar las dificultades inherentes a los sistemas de expresión procariote, por esto es esencial delimitar los alcances que pueden tener las bacterias más empleadas y aquellas que podrían representar alternativas prometedoras para la producción de proteínas de interés. La presente revisión bibliográfica está dirigida a realizar un estudio del estado del arte y de la técnica de los sistemas de expresión de proteínas heterólogas que existen en torno al empleo de bacterias, con impacto en la industria farmacéutica y biotecnológica.

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Protein Complex Production in Alternative Prokaryotic Hosts

Gómez

López-Estepa

Fernández

et al. 2016

Advanced Technologies for Protein Complex Production and Characterization

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Research for multiprotein expression in nonconventional bacterial and archaeal expression systems aims to exploit particular properties of "alternative" prokaryotic hosts that might make them more efficient than E. coli for particular applications, especially in those areas where more conventional bacterial hosts traditionally do not perform well. Currently, a wide range of products with clinical or industrial application have to be isolated from their native source, often microorganisms whose growth present numerous problems owing to very slow growth phenotypes or because they are unculturable under laboratory conditions. In those cases, transfer of the gene pathway responsible for synthesizing the product of interest into a suitable recombinant host becomes an attractive alternative solution. Despite many efforts dedicated to improving E. coli systems due to low cost, ease of use, and its dominant position as a ubiquitous expression host model, many alternative prokaryotic systems have been developed for heterologous protein expression mostly for biotechnological applications. Continuous research has led to improvements in expression yield through these non-conventional models, including Pseudomonas, Streptomyces and Mycobacterium as alternative bacterial expression hosts. Advantageous properties shared by these systems include low costs, high levels of secreted protein products and their safety of use, with non-pathogenic strains been commercialized. In addition, the use of extremophilic and halotolerant archaea as expression hosts has to be considered as a potential tool for the production of mammalian membrane proteins such as GPCRs.

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Heterologous production of Escherichia coli penicillin G acylase in Pseudomonas aeruginosa

Cited by 14 publications

References 61 publications

Mutations in the translation initiation region of the pac gene resulting in increased levels of activity of penicillin G acylase

Mutations in the translation initiation region of the pac gene resulting in increased levels of activity of penicillin G acylase

Las bacterias como sistema de expresión de proteínas heterólogas terapéuticas: una revisión bibliográfica

Protein Complex Production in Alternative Prokaryotic Hosts

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