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      A review on Lactococcus lactis: from food to factory

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          Abstract

          Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features of this gram-positive lactic acid non-colonizing gut bacteria include its generally recognized as safe (GRAS) status, probiotic properties, the absence of inclusion bodies and endotoxins, surface display and extracellular secretion technology, and a diverse selection of cloning and inducible expression vectors. This have made L. lactis a desirable and promising host on par with other well established model bacterial or yeast systems such as Escherichia coli, Salmonella cerevisiae and Bacillus subtilis. In this article, we review recent technological advancements, challenges, future prospects and current diversified examples on the use of L. lactis as a microbial cell factory. Additionally, we will also highlight latest medical-based applications involving whole-cell L. lactis as a live delivery vector for the administration of therapeutics against both communicable and non-communicable diseases.

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          Most cited references151

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          A phase I trial with transgenic bacteria expressing interleukin-10 in Crohn's disease.

          The use of living, genetically modified bacteria is an effective approach for topical delivery of immunomodulatory proteins. This strategy circumvents systemic side effects and allows long-term treatment of chronic diseases. However, treatment of patients with a living, genetically modified bacterium raises questions about the safety for human subjects per se and the biologic containment of the transgene. We treated Crohn's disease patients with genetically modified Lactococcus lactis (LL-Thy12) in which the thymidylate synthase gene was replaced with a synthetic sequence encoding mature human interleukin-10. Ten patients were included in a placebo-uncontrolled trial. Patients were assessed daily for the presence of potential adverse effects by direct questioning and assessment of disease activity. We evaluated the presence and kinetics of LL-Thy12 release in the stool of patients by conventional culturing and quantitative polymerase chain reaction of LL-Thy12 gene sequences. Treatment with LL-Thy12 was safe because only minor adverse events were present, and a decrease in disease activity was observed. Moreover, fecally recovered LL-Thy12 bacteria were dependent on thymidine for growth and interleukin-10 production, indicating that the containment strategy was effective. Here we show that the use of genetically modified bacteria for mucosal delivery of proteins is a feasible strategy in human beings. This novel strategy avoids systemic side effects and is biologically contained; therefore it is suitable as maintenance treatment for chronic intestinal disease.
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            10 years of the nisin-controlled gene expression system (NICE) in Lactococcus lactis.

            Lactococcus lactis is a Gram-positive lactic acid bacterium that, in addition to its traditional use in food fermentations, is increasingly used in modern biotechnological applications. In the last 25 years great progress has been made in the development of genetic engineering tools and the molecular characterization of this species. A new versatile and tightly controlled gene expression system, based on the auto-regulation mechanism of the bacteriocin nisin, was developed 10 years ago-the NIsin Controlled gene Expression system, called NICE. This system has become one of the most successful and widely used tools for regulated gene expression in Gram-positive bacteria. The review describes, after a brief introduction of the host bacterium L. lactis, the fundaments, components and function of the NICE system. Furthermore, an extensive overview is provided of the different applications in lactococci and other Gram-positive bacteria: (1) over-expression of homologous and heterologous genes for functional studies and to obtain large quantities of specific gene products, (2) metabolic engineering, (3) expression of prokaryotic and eukaryotic membrane proteins, (4) protein secretion and anchoring in the cell envelope, (5) expression of genes with toxic products and analysis of essential genes and (6) large-scale applications. Finally, an overview is given of growth and induction conditions for lab-scale and industrial-scale applications.
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              Treatment of murine colitis by Lactococcus lactis secreting interleukin-10.

              The cytokine interleukin-10 (IL-10) has shown promise in clinical trials for treatment of inflammatory bowel disease (IBD). Using two mouse models, we show that the therapeutic dose of IL-10 can be reduced by localized delivery of a bacterium genetically engineered to secrete the cytokine. Intragastric administration of IL-10-secreting Lactococcus lactis caused a 50% reduction in colitis in mice treated with dextran sulfate sodium and prevented the onset of colitis in IL-10(-/-) mice. This approach may lead to better methods for cost-effective and long-term management of IBD in humans.
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                Author and article information

                Contributors
                adelene@upm.edu.my
                lionelin@ucsiuniversity.edu.my
                erinlim@perdanauniveristy.edu.my
                raha@upm.edu.my
                Journal
                Microb Cell Fact
                Microb. Cell Fact
                Microbial Cell Factories
                BioMed Central (London )
                1475-2859
                4 April 2017
                4 April 2017
                2017
                : 16
                : 55
                Affiliations
                [1 ]GRID grid.11142.37, Department of Microbiology, Faculty of Biotechnology & Biomolecular Sciences, , University Putra Malaysia, ; 43400 Serdang, Selangor Malaysia
                [2 ]GRID grid.444472.5, Functional Food Research Group, Department of Biotechnology, Faculty of Applied Sciences, , UCSI University, ; Kuala Lumpur, Malaysia
                [3 ]GRID grid.261834.a, , Perdana University-Royal College of Surgeons in Ireland, Perdana University, ; Block B and D, MAEPS Building, MARDI Complex, Jalan MAEPS Perdana, 43400 Serdang, Selangor Malaysia
                [4 ]GRID grid.11142.37, Department of Cell & Molecular Biology, Faculty of Biotechnology & Biomolecular Sciences, , University Putra Malaysia, ; Serdang, Selangor Malaysia
                Article
                669
                10.1186/s12934-017-0669-x
                5379754
                28376880
                7dbbf75c-6e8b-4cd4-af5c-d1afea6b6829
                © The Author(s) 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 13 November 2016
                : 28 March 2017
                Funding
                Funded by: MOSTI
                Award ID: 02-02-22-SF0011
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100003093, Ministry of Higher Education, Malaysia;
                Award ID: FRGS/1/2014/SG05/UCSI/03/1
                Award Recipient :
                Categories
                Review
                Custom metadata
                © The Author(s) 2017

                Biotechnology
                lactococcus lactis,heterologous proteins,recombinant proteins,expression systems,secretion,surface display,microbial cell factory

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