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  1. Article ; Online: Engineering cyanobacterial chassis for improved electron supply toward a heterologous ene-reductase.

    Spasic, Jelena / Oliveira, Paulo / Pacheco, Catarina / Kourist, Robert / Tamagnini, Paula

    Journal of biotechnology

    2022  Volume 360, Page(s) 152–159

    Abstract: Cyanobacteria are noteworthy hosts for industrially relevant redox reactions, owing to a light-driven cofactor recycling system using water as electron donor. Customizing Synechocystis sp. PCC 6803 chassis by redirecting electron flow offers a ... ...

    Abstract Cyanobacteria are noteworthy hosts for industrially relevant redox reactions, owing to a light-driven cofactor recycling system using water as electron donor. Customizing Synechocystis sp. PCC 6803 chassis by redirecting electron flow offers a particularly interesting approach to further improve light-driven biotransformations. Therefore, different chassis expressing the heterologous ene-reductase YqjM (namely ΔhoxYH, Δflv3, ΔndhD2 and ΔhoxYHΔflv3) were generated/evaluated. The results showed the robustness of the chassis, that exhibited growth and oxygen evolution rates similar to Synechocystis wild-type, even when expressing YqjM. By engineering the electron flow, the YqjM light-driven stereoselective reduction of 2-methylmaleimide to 2-methylsuccinimide was significantly enhanced in all chassis. In the best performing chassis (ΔhoxYH, lacking an active bidirectional hydrogenase) a 39 % increase was observed, reaching an in vivo specific activity of 116 U g
    MeSH term(s) Oxidoreductases ; Electrons ; Oxygen
    Chemical Substances Oxidoreductases (EC 1.-) ; Oxygen (S88TT14065)
    Language English
    Publishing date 2022-11-09
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 843647-2
    ISSN 1873-4863 ; 0168-1656 ; 1389-0352
    ISSN (online) 1873-4863
    ISSN 0168-1656 ; 1389-0352
    DOI 10.1016/j.jbiotec.2022.11.005
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 2299: Show issues Location:
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  2. Article ; Online: Looking Outwards: Isolation of Cyanobacterial Released Carbohydrate Polymers and Proteins.

    Flores, Carlos / Tamagnini, Paula

    Journal of visualized experiments : JoVE

    2019  , Issue 147

    Abstract: Cyanobacteria can actively secrete a wide range of biomolecules into the extracellular environment, such as heteropolysaccharides and proteins. The identification and characterization of these biomolecules can improve knowledge about their secretion ... ...

    Abstract Cyanobacteria can actively secrete a wide range of biomolecules into the extracellular environment, such as heteropolysaccharides and proteins. The identification and characterization of these biomolecules can improve knowledge about their secretion pathways and help to manipulate them. Furthermore, some of these biomolecules are also interesting in terms of biotechnological applications. Described here are two protocols for easy and rapid isolation of cyanobacterial released carbohydrate polymers and proteins. The method for isolation of released carbohydrate polymers is based on conventional precipitation techniques of polysaccharides in aqueous solutions using organic solvents. This method preserves the characteristics of the polymer and simultaneously avoids the presence of contaminants from cell debris and culture medium. At the end of the process, the lyophilized polymer is ready to be used or characterized or can be subjected to further rounds of purification, depending on the final intended use. Regarding the isolation of the cyanobacterial exoproteome, the technique is based on the concentration of the cell-free medium after removal of the major contaminants by centrifugation and filtration. This strategy allows for reliable isolation of proteins that reach the extracellular milieu via membrane transporters or outer membrane vesicles. These proteins can be subsequently identified using standard mass spectrometry techniques. The protocols presented here can be applied not only to a wide range of cyanobacteria, but also to other bacterial strains. Furthermore, these procedures can be easily tailored according to the final use of the products, purity degree required, and bacterial strain.
    MeSH term(s) Carbohydrates/isolation & purification ; Cyanobacteria/metabolism ; Filtration ; Polymers/isolation & purification ; Proteins/isolation & purification
    Chemical Substances Carbohydrates ; Polymers ; Proteins
    Language English
    Publishing date 2019-05-27
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Video-Audio Media
    ZDB-ID 2259946-0
    ISSN 1940-087X ; 1940-087X
    ISSN (online) 1940-087X
    ISSN 1940-087X
    DOI 10.3791/59590
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Development of an ecotoxicological test procedure for soil microalgae.

    Carvalhido, Vânia / Bessa da Silva, Márcia / Santos, Marina / Tamagnini, Paula / Melo, Paula / Pereira, Ruth

    The Science of the total environment

    2021  Volume 783, Page(s) 147006

    Abstract: Since the 80s, ISO and OECD organizations have been developing guidelines for assessing the toxicity of new and existing chemical substances to soil biota. Up to now, any of these guidelines had soil algae as test organisms. Nevertheless, microalgae are ... ...

    Abstract Since the 80s, ISO and OECD organizations have been developing guidelines for assessing the toxicity of new and existing chemical substances to soil biota. Up to now, any of these guidelines had soil algae as test organisms. Nevertheless, microalgae are relevant components of soil microbial communities and soil biological crusts (BSC) with a great contribution to different soil functions and ecosystem services. In an attempt to bridge the gap, the present work aimed to develop, describe and validate a standard operating procedure for an ecotoxicological test with soil microalgae. Three phases were performed, each one with specific objectives. First, soil microalgae and cyanobacteria were isolated from BSC and then genetically and morphologically characterized. The green microalga Micractinium inermum was selected because it is a species with a wide geographic distribution. Secondly, M. inermum growth curves were obtained in liquid (BG
    MeSH term(s) Ecosystem ; Ecotoxicology ; Microalgae ; Reproducibility of Results ; Soil
    Chemical Substances Soil
    Language English
    Publishing date 2021-04-10
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 121506-1
    ISSN 1879-1026 ; 0048-9697
    ISSN (online) 1879-1026
    ISSN 0048-9697
    DOI 10.1016/j.scitotenv.2021.147006
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 949: Show issues Location:
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  4. Article ; Online: Assessing the Antitumor Potential of Variants of the Extracellular Carbohydrate Polymer from

    Mota, Rita / Lima, Raquel T / Flores, Carlos / Silva, Juliana F / Cruz, Beatriz / Alves, Bárbara / Pinto, Marta T / Adessi, Alessandra / Pereira, Sara B / De Philippis, Roberto / Soares, Paula / Tamagnini, Paula

    Polymers

    2023  Volume 15, Issue 6

    Abstract: Cancer is a leading cause of death worldwide with a huge societal and economic impact. Clinically effective and less expensive anticancer agents derived from natural sources can help to overcome limitations and negative side effects of chemotherapy and ... ...

    Abstract Cancer is a leading cause of death worldwide with a huge societal and economic impact. Clinically effective and less expensive anticancer agents derived from natural sources can help to overcome limitations and negative side effects of chemotherapy and radiotherapy. Previously, we showed that the extracellular carbohydrate polymer of a
    Language English
    Publishing date 2023-03-10
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2527146-5
    ISSN 2073-4360 ; 2073-4360
    ISSN (online) 2073-4360
    ISSN 2073-4360
    DOI 10.3390/polym15061382
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Untargeted Lipidomics Analysis of the Cyanobacterium

    Hewelt-Belka, Weronika / Kot-Wasik, Ágata / Tamagnini, Paula / Oliveira, Paulo

    International journal of molecular sciences

    2020  Volume 21, Issue 23

    Abstract: Cyanobacteria play an important role in several ecological environments, and they are widely accepted to be the ancestors of chloroplasts in modern plants and green algae. Cyanobacteria have become attractive models for metabolic engineering, with the ... ...

    Abstract Cyanobacteria play an important role in several ecological environments, and they are widely accepted to be the ancestors of chloroplasts in modern plants and green algae. Cyanobacteria have become attractive models for metabolic engineering, with the goal of exploring them as microbial cell factories. However, the study of cyanobacterial lipids' composition and variation, and the assessment of the lipids' functional and structural roles have been largely overlooked. Here, we aimed at expanding the cyanobacterial lipidomic analytical pipeline by using an untargeted lipidomics approach. Thus, the lipid composition variation of the model cyanobacterium
    MeSH term(s) Cyanobacteria/genetics ; Cyanobacteria/metabolism ; Fatty Acids/genetics ; Gene Deletion ; Gene Expression Regulation, Bacterial/genetics ; Lipidomics ; Lipids/genetics ; Membrane Lipids/genetics ; Metabolic Engineering ; Photosynthesis/genetics
    Chemical Substances Fatty Acids ; Lipids ; Membrane Lipids
    Language English
    Publishing date 2020-11-24
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms21238883
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article: Extracellular Vesicles: An Overlooked Secretion System in Cyanobacteria.

    Lima, Steeve / Matinha-Cardoso, Jorge / Tamagnini, Paula / Oliveira, Paulo

    Life (Basel, Switzerland)

    2020  Volume 10, Issue 8

    Abstract: In bacteria, the active transport of material from the interior to the exterior of the cell, or secretion, represents a very important mechanism of adaptation to the surrounding environment. The secretion of various types of biomolecules is mediated by a ...

    Abstract In bacteria, the active transport of material from the interior to the exterior of the cell, or secretion, represents a very important mechanism of adaptation to the surrounding environment. The secretion of various types of biomolecules is mediated by a series of multiprotein complexes that cross the bacterial membrane(s), each complex dedicated to the secretion of specific substrates. In addition, biological material may also be released from the bacterial cell in the form of vesicles. Extracellular vesicles (EVs) are bilayered, nanoscale structures, derived from the bacterial cell envelope, which contain membrane components as well as soluble products. In cyanobacteria, the knowledge regarding EVs is lagging far behind compared to what is known about, for example, other Gram-negative bacteria. Here, we present a summary of the most important findings regarding EVs in Gram-negative bacteria, discussing aspects of their composition, formation processes and biological roles, and highlighting a number of technological applications tested. This lays the groundwork to raise awareness that the release of EVs by cyanobacteria likely represents an important, and yet highly disregarded, survival strategy. Furthermore, we hope to motivate future studies that can further elucidate the role of EVs in cyanobacterial cell biology and physiology.
    Language English
    Publishing date 2020-07-31
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2662250-6
    ISSN 2075-1729
    ISSN 2075-1729
    DOI 10.3390/life10080129
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article: The Role of Outer Membrane Protein(s) Harboring SLH/OprB-Domains in Extracellular Vesicles' Production in

    Cardoso, Delfim / Lima, Steeve / Matinha-Cardoso, Jorge / Tamagnini, Paula / Oliveira, Paulo

    Plants (Basel, Switzerland)

    2021  Volume 10, Issue 12

    Abstract: Cyanobacteria are a group of photosynthetic prokaryotes that contribute to primary production on a global scale. These microorganisms release vesicles to the extracellular environment, spherical nanosized structures, derived essentially from the outer ... ...

    Abstract Cyanobacteria are a group of photosynthetic prokaryotes that contribute to primary production on a global scale. These microorganisms release vesicles to the extracellular environment, spherical nanosized structures, derived essentially from the outer membrane. Even though earlier works in model Gram-negative bacteria have hypothesized that outer membrane stability is crucial in vesicle formation, the mechanisms determining vesicle biogenesis in cyanobacteria remain unknown. Here, we report on the identification of six candidate genes encoding outer membrane proteins harboring SLH/OprB-domains in the genome of the model cyanobacterium
    Language English
    Publishing date 2021-12-14
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2704341-1
    ISSN 2223-7747
    ISSN 2223-7747
    DOI 10.3390/plants10122757
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article: CRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium

    Santos, Marina / Pacheco, Catarina C / Yao, Lun / Hudson, Elton P / Tamagnini, Paula

    Life (Basel, Switzerland)

    2021  Volume 11, Issue 11

    Abstract: The use of the versatile cyanobacterial extracellular polymeric substances (EPS) for biotechnological/biomedical applications implies an extensive knowledge of their biosynthetic pathways to improve/control polymer production yields and characteristics. ... ...

    Abstract The use of the versatile cyanobacterial extracellular polymeric substances (EPS) for biotechnological/biomedical applications implies an extensive knowledge of their biosynthetic pathways to improve/control polymer production yields and characteristics. The multiple copies of EPS-related genes, scattered throughout cyanobacterial genomes, adds another layer of complexity, making these studies challenging and time-consuming. Usually, this issue would be tackled by generating deletion mutants, a process that in cyanobacteria is also hindered by the polyploidy. Thus, the use of the CRISPRi multiplex system constitutes an efficient approach to addressing this redundancy. Here, three putative
    Language English
    Publishing date 2021-11-06
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2662250-6
    ISSN 2075-1729
    ISSN 2075-1729
    DOI 10.3390/life11111198
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  9. Article ; Online: The role of positive charged residue in the proton-transfer mechanism of two-domain laccase from

    Gabdulkhakov, Azat / Kolyadenko, Ilya / Oliveira, Paulo / Tamagnini, Paula / Mikhaylina, Alisa / Tishchenko, Svetlana

    Journal of biomolecular structure & dynamics

    2021  Volume 40, Issue 18, Page(s) 8324–8331

    Abstract: Multi-copper oxidases are capable of coupling the one-electron oxidation of four substrate equivalents to the four-electron reduction of dioxygen to two molecules of water. This process takes place at the trinuclear copper center of the enzymes. ... ...

    Abstract Multi-copper oxidases are capable of coupling the one-electron oxidation of four substrate equivalents to the four-electron reduction of dioxygen to two molecules of water. This process takes place at the trinuclear copper center of the enzymes. Previously, the main catalytic stages for three-domain (3D) laccases have been identified. However, for bacterial small two-domain (2D) laccases several questions remain to be answered. One of them is the nature of the protonation events upon the reductive cleavage of dioxygen to water. In 3D laccases, acidic residues play a key role in the protonation mechanisms. In this study, the role of the Arg240 residue, located within the T2 tunnel of 2D laccase from
    MeSH term(s) Laccase/chemistry ; Laccase/metabolism ; Oxidation-Reduction ; Oxygen ; Protons ; Streptomyces ; Water
    Chemical Substances Protons ; Water (059QF0KO0R) ; Laccase (EC 1.10.3.2) ; Oxygen (S88TT14065)
    Language English
    Publishing date 2021-04-19
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 49157-3
    ISSN 1538-0254 ; 0739-1102
    ISSN (online) 1538-0254
    ISSN 0739-1102
    DOI 10.1080/07391102.2021.1911852
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 2093: Show issues Location:
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  10. Article: Heterologous Production of Glycine Betaine Using

    Ferreira, Eunice A / Pacheco, Catarina C / Rodrigues, João S / Pinto, Filipe / Lamosa, Pedro / Fuente, David / Urchueguía, Javier / Tamagnini, Paula

    Frontiers in bioengineering and biotechnology

    2022  Volume 9, Page(s) 821075

    Abstract: Among compatible solutes, glycine betaine has various applications in the fields of nutrition, pharmaceuticals, and cosmetics. Currently, this compound can be extracted from sugar beet plants or obtained by chemical synthesis, resulting in low yields or ... ...

    Abstract Among compatible solutes, glycine betaine has various applications in the fields of nutrition, pharmaceuticals, and cosmetics. Currently, this compound can be extracted from sugar beet plants or obtained by chemical synthesis, resulting in low yields or high carbon footprint, respectively. Hence, in this work we aimed at exploring the production of glycine betaine using the unicellular cyanobacterium
    Language English
    Publishing date 2022-01-07
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2719493-0
    ISSN 2296-4185
    ISSN 2296-4185
    DOI 10.3389/fbioe.2021.821075
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