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  • 1.
    Gustavsson, Martin
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Hörnström, David
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Lundh, Susanna
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Belotserkovsky, Jaroslav
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Larsson, Gen
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Biocatalysis on the surface of Escherichia coli: melanin pigmentation of the cell exterior2016Inngår i: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 6, artikkel-id 36117Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Today, it is considered state-of-the-art to engineer living organisms for various biotechnology applications. Even though this has led to numerous scientific breakthroughs, the enclosed interior of bacterial cells still restricts interactions with enzymes, pathways and products due to the mass-transfer barrier formed by the cell envelope. To promote accessibility, we propose engineering of biocatalytic reactions and subsequent product deposition directly on the bacterial surface. As a proof-of-concept, we used the AIDA autotransporter vehicle for Escherichia coli surface expression of tyrosinase and fully oxidized externally added tyrosine to the biopolymer melanin. This resulted in a color change and creation of a black cell exterior. The capture of ninety percent of a pharmaceutical wastewater pollutant followed by regeneration of the cell bound melanin matrix through a simple pH change, shows the superior function and facilitated processing provided by the surface methodology. The broad adsorption spectrum of melanin could also allow removal of other micropollutants.

  • 2.
    Jarmander, Johan
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Belotserkovsky, Jaroslav
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Sjöberg, Gustav
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Guevara-Martínez, Mónica
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi. Universidad Mayor de San Simón, Bolivia .
    Zabaleta, Mariel Perez
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi. Universidad Mayor de San Simón, Bolivia .
    Quillaguaman, Jorge
    Universidad Mayor de San Simón, Bolivia .
    Larsson, Gen
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Cultivation strategies for production of (R)-3-hydroxybutyric acid from simultaneous consumption of glucose, xylose and arabinose by Escherichia coli2015Inngår i: Microbial Cell Factories, ISSN 1475-2859, E-ISSN 1475-2859, Vol. 14, nr 1, s. 51-Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Background

    Lignocellulosic waste is a desirable biomass for use in second generation biorefineries. Up to 40 % of its sugar content consist of pentoses, which organisms either take up sequentially after glucose depletion, or not at all. A previously described Escherichia coli strain, PPA652ara, capable of simultaneous consumption of glucose, xylose and arabinose was in the present work utilized for production of (R)-3-hydroxybutyric acid (3HB) from a mixture of glucose, xylose and arabinose.

    Results

    The Halomonas boliviensis genes for 3HB production were for the first time cloned into E. coli PPA652ara leading to product secretion directly into the medium. Process design was based on comparisons of batch, fed-batch and continuous cultivation, where both excess and limitation of the carbon mixture was studied. Carbon limitation resulted in low specific productivity of 3HB (< 2 mg g-1 h-1) compared to carbon excess (25 mg g-1 h-1), but the yield of 3HB/cell dry weight (Y3HB/CDW) was very low (0.06 g g-1)during excess. Nitrogen-exhausted conditions could be used to sustain a high specific productivity (31 mg g-1 h-1) and to increase the yield of 3HB/cell dry weight to 1.38 g g-1. Nitrogen-limited fed-batch process design lead to further increased specific productivity (38 mg g-1 h-1) but also to additional cell growth (Y3HB/CDW = 0.16 g g-1). Strain PPA652ara did under all processing conditions simultaneously consume glucose, xylose and arabinose, which was not the case for a reference wild type E. coli, which also gave a higher carbon flux to acetic acid.

    Conclusions

    It was demonstrated that by using the strain E. coli PPA652ara it was possible to design a production process for 3HB from a mixture of glucose, xylose and arabinose where all sugars were consumed. An industrial 3HB production process is proposed to be divided into a growth and a production phase, and nitrogen depletion/limitation is a potential strategy to maximize the yield of 3HB/CDW in the latter. The specific productivity of 3HB by E. coli reported here from glucose, xylose and arabinose is further comparable to the current state of the art for production of 3HB from glucose sources.

  • 3.
    Jarmander, Johan
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Guevara, Mónica
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Zabaleta, Mariel Perez
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Sjöberg, Gustav
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Belotserkovsky, Jaroslav
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Quillaguaman, Jorge
    Larsson, Gen
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Production of 3-hydroxybutyrate from waste biomass by metabolically engineered Escherichia coli2014Inngår i: New Biotechnology, ISSN 1871-6784, E-ISSN 1876-4347, Vol. 31, s. S94-S95Artikkel i tidsskrift (Annet vitenskapelig)
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