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      Whole cell biocatalysts: essential workers from Nature to the industry

      review-article
      1 ,
      Microbial Biotechnology
      John Wiley and Sons Inc.

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          Summary

          Microorganisms have been exposed to a myriad of substrates and environmental conditions throughout evolution resulting in countless metabolites and enzymatic activities. Although mankind have been using these properties for centuries, we have only recently learned to control their production, to develop new biocatalysts with high stability and productivity and to improve their yields under new operational conditions. However, microbial cells still provide the best known environment for enzymes, preventing conformational changes in the protein structure in non‐conventional medium and under harsh reaction conditions, while being able to efficiently regenerate necessary cofactors and to carry out cascades of reactions. Besides, a still unknown microbe is probably already producing a compound that will cure cancer, Alzeihmer's disease or kill the most resistant pathogen. In this review, the latest developments in screening desirable activities and improving production yields are discussed.

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          Most cited references63

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          Big effects from small changes: possible ways to explore nature's chemical diversity.

          Fungi or bacteria that produce secondary metabolites often have the potential to bring up various compounds from a single strain. The molecular basis for this well-known observation was confirmed in the last few years by several sequencing projects of different microorganisms. Besides well-known examples about induction of a selected biosynthesis (for example, by high- or low-phosphate cultivation media), no overview about the potential in this field for finding natural products was given. We have investigated the systematic alteration of easily accessible cultivation parameters (for example, media composition, aeration, culture vessel, addition of enzyme inhibitors) in order to increase the number of secondary metabolites available from one microbial source. We termed this way of revealing nature's chemical diversity the 'OSMAC (One Strain-Many Compounds) approach' and by using it we were able to isolate up to 20 different metabolites in yields up to 2.6 g L(-1) from a single organism. These compounds cover nearly all major natural product families, and in some cases the high production titer opens new possibilities for semisynthetic methods to enhance even more the chemical diversity of selected compounds. The OSMAC approach offers a good alternative to industrial high-throughput screening that focuses on the active principle in a distinct bioassay. In consequence, the detection of additional compounds that might be of interest as lead structures in further bioassays is impossible and clearly demonstrates the deficiency of the industrial procedure. Furthermore, our approach seems to be a useful tool to detect those metabolites that are postulated to be the final products of an amazing number of typical secondary metabolite gene clusters identified in several microorganisms. If one assumes a (more or less) defined reservoir of genetic possibilities for several biosynthetic pathways in one strain that is used for a highly flexible production of secondary metabolites depending on the environment, the OSMAC approach might give more insight into the role of secondary metabolism in the microbial community or during the evolution of life itself.
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            Industrial biocatalysis today and tomorrow.

            The use of biocatalysis for industrial synthetic chemistry is on the verge of significant growth. Biocatalytic processes can now be carried out in organic solvents as well as aqueous environments, so that apolar organic compounds as well as water-soluble compounds can be modified selectively and efficiently with enzymes and biocatalytically active cells. As the use of biocatalysis for industrial chemical synthesis becomes easier, several chemical companies have begun to increase significantly the number and sophistication of the biocatalytic processes used in their synthesis operations.
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              Cultivating the uncultured.

              The recent application of molecular phylogeny to environmental samples has resulted in the discovery of an abundance of unique and previously unrecognized microorganisms. The vast majority of this microbial diversity has proved refractory to cultivation. Here, we describe a universal method that provides access to this immense reservoir of untapped microbial diversity. This technique combines encapsulation of cells in gel microdroplets for massively parallel microbial cultivation under low nutrient flux conditions, followed by flow cytometry to detect microdroplets containing microcolonies. The ability to grow and study previously uncultured organisms in pure culture will enhance our understanding of microbial physiology and metabolic adaptation and will provide new sources of microbial metabolites. We show that this technology can be applied to samples from several different environments, including seawater and soil.
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                Author and article information

                Contributors
                ccarvalho@tecnico.ulisboa.pt
                Journal
                Microb Biotechnol
                Microb Biotechnol
                10.1111/(ISSN)1751-7915
                MBT2
                Microbial Biotechnology
                John Wiley and Sons Inc. (Hoboken )
                1751-7915
                03 May 2016
                March 2017
                : 10
                : 2 ( doiID: 10.1111/mbt2.2017.10.issue-2 )
                : 250-263
                Affiliations
                [ 1 ] iBB‐Institute for Bioengineering and Biosciences Department of Bioengineering Instituto Superior TécnicoUniversidade de Lisboa Av. Rovisco Pais Lisbon 1049‐001Portugal
                Author notes
                [*] [* ]For correspondence. E‐mail: ccarvalho@ 123456tecnico.ulisboa.pt ; Tel. +351 21 841 95 94; Fax +351 21 841 90 62.
                Article
                MBT212363
                10.1111/1751-7915.12363
                5328830
                27145540
                f69b9bf6-1603-43c7-9b77-81e4cdcb370c
                © 2016 The Author. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

                This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 27 January 2016
                : 28 March 2016
                : 31 March 2016
                Page count
                Figures: 1, Tables: 2, Pages: 14, Words: 10597
                Funding
                Funded by: ʻFCT Investigator 2013ʼ
                Award ID: IF/01203/2013/CP1163/CT0002
                Funded by: European Union's Horizon 2020 Research and Innovation Programme
                Award ID: 634486
                Categories
                Minireview
                Minireviews
                Custom metadata
                2.0
                mbt212363
                March 2017
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.0.7 mode:remove_FC converted:27.02.2017

                Biotechnology
                Biotechnology

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