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β-谷甾醇在小型设备中的生物转化:从微量滴定板到微流控反应器。

β-Sitosterol Bioconversion in Small-Scale Devices: From Microtiter Plates to Microfluidic Reactors.

机构信息

Department of Biochemical Engineering, University College London, London, UK.

iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.

出版信息

Methods Mol Biol. 2023;2704:201-219. doi: 10.1007/978-1-0716-3385-4_12.

Abstract

Small-scale devices are routinely used as low-cost miniaturized bioreactors due to the large number of experiments that can be conducted simultaneously under similar conditions and replicate all functions of bench-scale reactors at dramatically smaller volumes. Microtiter plates, due to the standard footprint, can be integrated with liquid handling systems and associated equipment, expanding considerably their application and use. However, care has to be taken to operate the microtiter plates in optimized mixing and oxygen transfer conditions, preventing medium evaporation in prolonged experiment runs. Recently, to increase data quality, microbioreactors have emerged as an alternative to shaken systems. These systems offer higher degree of control over key process variables and when combined with sensing technology increase dramatically the reliability of translational process data. In this chapter, we describe the production of 4-androstene-3,17-dione (androstenedione (AD)), a key pharmaceutical steroid intermediate, by Mycobacterium sp. NRRL B-3805 via the selective cleavage of the side-chain of β-sitosterol using 24-well microtiter plates and microfluidic microbioreactors.

摘要

由于可以在相似条件下同时进行大量实验,并在更小的体积中复制台式反应器的所有功能,因此小规模设备通常被用作低成本的微型生物反应器。由于标准的占地面积,微孔板可以与液体处理系统和相关设备集成,从而大大扩展了它们的应用和用途。但是,必须注意在优化的混合和氧气转移条件下操作微孔板,以防止在长时间的实验运行中介质蒸发。最近,为了提高数据质量,微生物反应器已成为摇床系统的替代品。这些系统可以对关键过程变量进行更高程度的控制,并且与传感技术结合使用时,会极大地提高转化过程数据的可靠性。在本章中,我们描述了使用 24 孔微量滴定板和微流控微生物反应器,通过分枝杆菌 NRRL B-3805 对 β-谷甾醇的侧链进行选择性切割来生产 4-雄烯二酮(雄烯二酮(AD)),这是一种关键的药物甾体中间体。

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