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<w:instrText>ADDIN CSL_CITATION { ... }</w:instrText>

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 {
    "citationItems": [{
        "id": "ITEM-1",
        "itemData": {
            "DOI": "10.1128/AEM.02591-07",
            "abstract": "Malic acid is a potential biomass-derivable \u201cbuilding block\u201d for chemical synthesis. Since wild-type Saccharomyces cerevisiae strains produce only low levels of malate, metabolic engineering is required to achieve efficient malate production with this yeast. A promising pathway for malate production from glucose proceeds via carboxylation of pyruvate, followed by reduction of oxaloacetate to malate. This redox- and ATP-neutral, CO2-fixing pathway has a theoretical maximum yield of 2 mol malate (mol glucose)\u22121. A previously engineered glucose-tolerant, C2-independent pyruvate decarboxylase-negative S. cerevisiae strain was used as the platform to evaluate the impact of individual and combined introduction of three genetic modifications: (i) overexpression of the native pyruvate carboxylase encoded by PYC2, (ii) high-level expression of an allele of the MDH3 gene, of which the encoded malate dehydrogenase was retargeted to the cytosol by deletion of the C-terminal peroxisomal targeting sequence, and (iii) functional expression of the Schizosaccharomyces pombe malate transporter gene SpMAE1. While single or double modifications improved malate production, the highest malate yields and titers were obtained with the simultaneous introduction of all three modifications. In glucose-grown batch cultures, the resulting engineered strain produced malate at titers of up to 59 g liter\u22121 at a malate yield of 0.42 mol (mol glucose)\u22121. Metabolic flux analysis showed that metabolite labeling patterns observed upon nuclear magnetic resonance analyses of cultures grown on 13C-labeled glucose were consistent with the envisaged nonoxidative, fermentative pathway for malate production. The engineered strains still produced substantial amounts of pyruvate, indicating that the pathway efficiency can be further improved.",
            "author": [{
                "dropping-particle": "",
                "family": "Zelle",
                "given": "Rintze M.",
                "non-dropping-particle": "",
                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Hulster",
                "given": "Erik",
                "non-dropping-particle": "de",
                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Winden",
                "given": "Wouter A.",
                "non-dropping-particle": "van",
                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Waard",
                "given": "Pieter",
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                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Dijkema",
                "given": "Cor",
                "non-dropping-particle": "",
                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Winkler",
                "given": "Aaron A.",
                "non-dropping-particle": "",
                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Geertman",
                "given": "Jan-Maarten A.",
                "non-dropping-particle": "",
                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Dijken",
                "given": "Johannes P.",
                "non-dropping-particle": "van",
                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Pronk",
                "given": "Jack T.",
                "non-dropping-particle": "",
                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Maris",
                "given": "Antonius J. A.",
                "non-dropping-particle": "van",
                "parse-names": false,
                "suffix": ""
            }],
            "container-title": "Applied and Environmental Microbiology",
            "id": "ITEM-1",
            "issue": "9",
            "issued": {
                "date-parts": [
                    ["2008"]
                ]
            },
            "page": "2766-2777",
            "title": "Malic Acid Production by Saccharomyces cerevisiae: Engineering of Pyruvate Carboxylation, Oxaloacetate Reduction, and Malate Export",
            "type": "article-journal",
            "volume": "74"
        },
        "uris": ["http://www.mendeley.com/documents/?uuid=23d11864-e841-3d60-8a56-32eefa603948"]
    }, {
        "id": "ITEM-2",
        "itemData": {
            "DOI": "10.1128/AEM.01077-10",
            "abstract": "Pyruvate carboxylase is the sole anaplerotic enzyme in glucose-grown cultures of wild-type Saccharomyces cerevisiae. Pyruvate carboxylase-negative (Pyc \u060a) S. cerevisiae strains cannot grow on glucose unless media are supplemented with C 4 compounds, such as aspartic acid. In several succinate-producing prokaryotes, phos-phoenolpyruvate carboxykinase (PEPCK) fulfills this anaplerotic role. However, the S. cerevisiae PEPCK encoded by PCK1 is repressed by glucose and is considered to have a purely decarboxylating and gluconeogenic function. This study investigates whether and under which conditions PEPCK can replace the anaplerotic function of pyruvate carboxylase in S. cerevisiae. Pyc \u060a S. cerevisiae strains constitutively overexpressing the PEPCK either from S. cerevisiae or from Actinobacillus succinogenes did not grow on glucose as the sole carbon source. However, evolutionary engineering yielded mutants able to grow on glucose as the sole carbon source at a maximum specific growth rate of ca. 0.14 h \u060a1 , one-half that of the (pyruvate carboxylase-positive) reference strain grown under the same conditions. Growth was dependent on high carbon dioxide concentrations, indicating that the reaction catalyzed by PEPCK operates near thermodynamic equilibrium. Analysis and reverse engineering of two independently evolved strains showed that single point mutations in pyruvate kinase, which competes with PEPCK for phosphoenolpyruvate, were sufficient to enable the use of PEPCK as the sole anaplerotic enzyme. The PEPCK reaction produces one ATP per carboxylation event, whereas the original route through pyruvate kinase and pyruvate carboxylase is ATP neutral. This increased ATP yield may prove crucial for engineering of efficient and low-cost anaerobic production of C 4 dicarboxylic acids in S. cerevisiae. Interest in biotechnological production of the four-carbon dicarboxylic acids fumarate, succinate, and malate from sugars has strongly increased in recent years (19), as these sugar-derived acids are seen as potential replacements for oil-derived chemical intermediates such as maleic anhydride (41). Meta-bolic engineering of Escherichia coli has resulted in strains capable of producing 73 g liter \u03ea1 succinate at pH 7.0 with a yield that, at 1.61 mol per mol glucose (21), is at 94% of the theoretical maximum. The crucial role of carboxylation reac-tions in this biotechnological process is illustrated by a carbon yield of 1.07 C mol succinate per C mol of glucose\u2026",
            "author": [{
                "dropping-particle": "",
                "family": "Zelle",
                "given": "Rintze M",
                "non-dropping-particle": "",
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                "dropping-particle": "",
                "family": "Trueheart",
                "given": "Josh",
                "non-dropping-particle": "",
                "parse-names": false,
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            }, {
                "dropping-particle": "",
                "family": "Harrison",
                "given": "Jacob C",
                "non-dropping-particle": "",
                "parse-names": false,
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            }, {
                "dropping-particle": "",
                "family": "Pronk",
                "given": "Jack T",
                "non-dropping-particle": "",
                "parse-names": false,
                "suffix": ""
            }, {
                "dropping-particle": "",
                "family": "Maris",
                "given": "Antonius J A",
                "non-dropping-particle": "van",
                "parse-names": false,
                "suffix": ""
            }],
            "container-title": "Applied and Environmental Microbiology",
            "id": "ITEM-2",
            "issue": "16",
            "issued": {
                "date-parts": [
                    ["2010"]
                ]
            },
            "page": "5383-5389",
            "title": "Phosphoenolpyruvate Carboxykinase as the Sole Anaplerotic Enzyme in Saccharomyces cerevisiae",
            "type": "article-journal",
            "volume": "76"
        },
        "uris": ["http://www.mendeley.com/documents/?uuid=1916c171-8e1e-3ad9-b16b-f99beef40f81"]
    }],
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    },
    "properties": {
        "noteIndex": 0
    },
    "schema": "https://github.com/citation-style-language/schema/raw/master/csl-citation.json"
 }

rmzelle / ref-extractor

Support Mendeley fields #3