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首页> 外文期刊>Bioprocess and Biosystems Engineering >Immobilized L-aspartate ammonia-lyase from Bacillus sp. YM55-1 as biocatalyst for highly concentrated L-aspartate synthesis
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Immobilized L-aspartate ammonia-lyase from Bacillus sp. YM55-1 as biocatalyst for highly concentrated L-aspartate synthesis

机译：固定的芽孢杆菌属的L-天冬氨酸氨裂合酶。 YM55-1作为高浓度L-天门冬氨酸合成的生物催化剂

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摘要

L-Aspartate ammonia-lyase from Bacillus sp. YM55-1 (AspB, EC 4.3.1.1) catalyzes the reversible conversion of L-aspartate (Asp) into fumarate and ammonia with a high specific activity toward the substrate. AspB was expressed in Escherichia coli and partially purified by heat precipitation and saturation with ammonium sulfate reaching purification factor of 7.7 and specific activity of 334 U/mg of protein. AspB was immobilized by covalent attachment on Eupergit® C (epoxy support) and MANA-agarose (amino support), and entrapment in LentiKats® (polyvinyl alcohol) with retained activities of 24, 85 and 63 %, respectively. Diffusional limitations were only observed for the enzyme immobilized in LentiKats® and were overcome by increasing substrate concentration. Free and immobilized AspB were used for the synthesis of aspartate achieving high product concentration (≥450 mM) after 24 h of reaction. Immobilized biocatalysts were efficiently reused in 5 cycles of Asp synthesis, keeping over 90 % of activity and reaching over 90 % of conversion in all the cases.
机译：来自芽孢杆菌属的L-天冬氨酸氨裂合酶。 YM55-1（AspB，EC 4.3.1.1）催化L-天冬氨酸（Asp）可逆转化为富马酸酯和氨，对底物具有很高的比活性。 AspB在大肠杆菌中表达，并通过热沉淀和硫酸铵饱和进行部分纯化，达到7.7的纯化因子和334 U / mg蛋白质的比活。通过共价键将AspB固定在Eupergit®C（环氧载体）和MANA-琼脂糖（氨基载体）上，并截留在LentiKats®（聚乙烯醇）中，保留的活性分别为24％，85％和63％。仅在固定在LentiKats®中的酶中观察到了扩散限制，并且可以通过增加底物浓度来克服。反应24小时后，游离的固定化AspB用于合成高产物浓度（≥450mM）的天冬氨酸。固定化的生物催化剂可在5个Asp合成循环中有效重复使用，在所有情况下均保持90％以上的活性并达到90％以上的转化率。

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  • 来源
    《Bioprocess and Biosystems Engineering》 |2012年第8期|p.1437-1444|共8页
  • 作者

    Max Cardenas-Fernandez; Carmen Lopez; Gregorio Alvaro; Josep Lopez-Santin;

  • 作者单位

    Applied Biocatalysis Unit associated to 1QAC (UAB-CSIC), Department of Chemical Engineering, School of Engineering,Universitat Autonoma de Barcelona, Bellaterra (Cerdanyola del Valles), 08193 Catalonia, Spain;

    Applied Biocatalysis Unit associated to 1QAC (UAB-CSIC), Department of Chemical Engineering, School of Engineering,Universitat Autonoma de Barcelona, Bellaterra (Cerdanyola del Valles), 08193 Catalonia, Spain;

    Applied Biocatalysis Unit associated to 1QAC (UAB-CSIC), Department of Chemical Engineering, School of Engineering,Universitat Autonoma de Barcelona, Bellaterra (Cerdanyola del Valles), 08193 Catalonia, Spain;

    Applied Biocatalysis Unit associated to 1QAC (UAB-CSIC), Department of Chemical Engineering, School of Engineering,Universitat Autonoma de Barcelona, Bellaterra (Cerdanyola del Valles), 08193 Catalonia, Spain;

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  • 正文语种 eng
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  • 关键词

    enzymatic L-aspartate synthesis; l-aspartate ammonia-lyase immobilization; eupergit® C; MANA-agarose; lentikats®;

    机译：酶促L-天冬氨酸合成;L-天冬氨酸氨解酶固定化;eupergit®C;MANA-琼脂糖;lentikats®;

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