Browsing by Author "Mulet, Ana Paula"

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    Bacteria-polymer composite material for glycerol valorization
    (2023) Ripoll, Magdalena; Soriano, Nicolás; Ibarburu, Sofía; Dalies, Malena; Mulet, Ana Paula; Betancor, Lorena
    Bacterial immobilization is regarded as an enabling technology to improve the stability and reusability of biocatalysts. Natural polymers are often used as immobilization matrices but present certain drawbacks, such as biocatalyst leakage and loss of physical integrity upon utilization in bioprocesses. Herein, we prepared a hybrid polymeric matrix that included silica nanoparticles for the unprecedented immobilization of the industrially relevant Gluconobacter frateurii (Gfr). This biocatalyst can valorize glycerol, an abundant by-product of the biodiesel industry, into glyceric acid (GA) and dihydroxyacetone (DHA). Different concentrations of siliceous nanosized materials, such as biomimetic Si nanoparticles (SiNps) and montmorillonite (MT), were added to alginate. These hybrid materials were significantly more resistant by texture analysis and presented a more compact structure as seen by scanning electron microscopy. The preparation including 4% alginate with 4% SiNps proved to be the most resistant material, with a homogeneous distribution of the biocatalyst in the beads as seen by confocal microscopy using a fluorescent mutant of Gfr. It produced the highest amounts of GA and DHA and could be reused for up to eight consecutive 24 h reactions with no loss of physical integrity and negligible bacterial leakage. Overall, our results indicate a new approach to generating biocatalysts using hybrid biopolymer supports.
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    CRISPR tools in bacterial whole-cell biocatalysis
    (2023) Mulet, Ana Paula; Ripoll, Magdalena; Betancor, Lorena
    Biocatalysis has emerged as a promising alternative to conventional chemical processes for the production of a wide range of chemicals, providing a sustainable solution to the problem of limited resources due to an ever-increasing global population. This approach involves the use of biobased catalysts, such as whole microorganisms or enzymes, to perform chemical conversions. While whole-cell biocatalysts offer advantages over the use of free enzymes, limitations related to productivity and undesired compound production have been observed when using microorganisms. Offering high specificity, broad applicability, and increased efficiency over traditional genetic engineering methods, CRISPR-based technologies may be the quintessential tool for the fit-for-purpose design of efficient bacterial biocatalysts. In this work, we aim to demonstrate the potential of CRISPR-based technologies to enhance whole-cell bacterial biotransformations for a more sustainable obtention of industrially important products. We have included a comprehensive and in-depth analysis of the current state of the art, emphasizing challenges and opportunities for future research. Through a critical analysis of reported examples, we intend to highlight the opportunities and advantages offered by CRISPR-based technologies in the field of biocatalysis for more efficient, sustainable, and translational processes.