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首页> 外文期刊>Bioprocess and Biosystems Engineering >Microscale methods to rapidly evaluate bioprocess options for increasing bioconversion yields: application to the ω-transaminase synthesis of chiral amines
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Microscale methods to rapidly evaluate bioprocess options for increasing bioconversion yields: application to the ω-transaminase synthesis of chiral amines

机译：快速评估生物工艺选择以提高生物转化产量的微型方法：在手性胺的ω-转氨酶合成中的应用

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摘要

This work aims to establish microscale methods to rapidly explore bioprocess options that might be used to enhance bioconversion reaction yields: either by shifting unfavourable reaction equilibria or by overcoming substrate and/or product inhibition. As a typical and industrially relevant example of the problems faced we have examined the asymmetric synthesis of (2S,3R)-2-amino-1,3,4-butanetriol from L-erythrulose using the ω-transaminase from Chromobacterium violaceum DSM30191 (CV2025 ω-Tam) and methylbenzylamine as the amino donor. The first process option involves the use of alternative amino donors. The second couples the CV2025 ω-Tam with alcohol dehydrogenase and glucose dehydrogenase for removal of the acetophenone (AP) byproduct by in situ conversion to ®-1-phenylethanol. The final approaches involve physical in-situ product removal methods. Reduced pressure conditions, attained using a 96-well vacuum manifold were used to selectively increase evaporation of the volatile AP while polymeric resins were also utilised for selective adsorption of AP from the bioconversion medium. For the particular reaction studied here the most promising bioprocess options were use of an alternative amino donor, such as isopropylamine, which enabled a 2.8-fold increase in reaction yield, or use of a second enzyme system which achieved a 3.3-fold increase in yield.
机译：这项工作旨在建立微观方法，以快速探索可用于提高生物转化反应产率的生物工艺选择：通过转移不利的反应平衡或克服底物和/或产物抑制作用。作为面临的问题的典型的与工业相关的例子，我们研究了使用紫堇杆菌DSM30191（CV2025）的ω-转氨酶从L-赤藓糖不对称合成（2S，3R）-2-氨基-1,3,4-丁三醇ω-Tam）和甲基苄胺作为氨基供体。第一个工艺选择涉及使用其他氨基供体。第二步将CV2025ω-Tam与醇脱氢酶和葡萄糖脱氢酶偶合，以通过原位转化为-1-苯乙醇去除苯乙酮（AP）副产物。最终方法涉及物理原位产物去除方法。使用96孔真空歧管达到的减压条件用于选择性地增加挥发性AP的蒸发，而聚合树脂还用于从生物转化介质中选择性吸附AP。对于此处研究的特定反应，最有前途的生物工艺选择是使用其他氨基供体（例如异丙胺），后者可使反应收率提高2.8倍，或使用第二种酶系统，可使收率提高3.3倍。

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  • 来源
    《Bioprocess and Biosystems Engineering》 |2014年第5期|931-941|共11页
  • 作者

    Murni Halim; Leonardo Rios-Solis; Martina Micheletti; John M. Ward; Gary J. Lye;

  • 作者单位

    Department of Biochemical Engineering, The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK,Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia;

    Department of Biochemical Engineering, The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK;

    Department of Biochemical Engineering, The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK;

    Department of Biochemical Engineering, The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK;

    Department of Biochemical Engineering, The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    ω-Transaminase; Asymmetric synthesis; Microscale bioprocessing; Equilibrium-controlled reaction; Product inhibition;

    机译：ω-转氨酶;不对称合成;微型生物加工;平衡控制反应;产品抑制;

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