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大肠杆菌赖氨酸脱羧酶的催化活性包涵体生产 1,5-二氨基戊烷。

Catalytically active inclusion bodies of L-lysine decarboxylase from E. coli for 1,5-diaminopentane production.

机构信息

Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, 52425, Jülich, Germany.

Bioeconomy Science Center (BioSC), c/o, Forschungszentrum Jülich, 52425, Jülich, Germany.

出版信息

Sci Rep. 2018 Apr 11;8(1):5856. doi: 10.1038/s41598-018-24070-2.

DOI:10.1038/s41598-018-24070-2
PMID:29643457
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5895699/
Abstract

Sustainable and eco-efficient alternatives for the production of platform chemicals, fuels and chemical building blocks require the development of stable, reusable and recyclable biocatalysts. Here we present a novel concept for the biocatalytic production of 1,5-diaminopentane (DAP, trivial name: cadaverine) using catalytically active inclusion bodies (CatIBs) of the constitutive L-lysine decarboxylase from E. coli (EcLDCc-CatIBs) to process L-lysine-containing culture supernatants from Corynebacterium glutamicum. EcLDCc-CatIBs can easily be produced in E. coli followed by a simple purification protocol yielding up to 43% dry CatIBs per dry cell weight. The stability and recyclability of EcLDCc-CatIBs was demonstrated in (repetitive) batch experiments starting from L-lysine concentrations of 0.1 M and 1 M. EcLDC-CatIBs exhibited great stability under reaction conditions with an estimated half-life of about 54 h. High conversions to DAP of 87-100% were obtained in 30-60 ml batch reactions using approx. 180-300 mg EcLDCc-CatIBs, respectively. This resulted in DAP titres of up to 88.4 g l and space-time yields of up to 660 g l d per gram dry EcLDCc-CatIBs. The new process for DAP production can therefore compete with the currently best fermentative process as described in the literature.

摘要

可持续且生态高效的平台化学品、燃料和化学构建块生产需要开发稳定、可重复使用和可回收的生物催化剂。在这里,我们提出了一种使用催化活性包含体(CatIBs)的新型概念,用于生物催化生产 1,5-二氨基戊烷(DAP,俗称 cadaverine),该 CatIBs 来自大肠杆菌中的组成型 L-赖氨酸脱羧酶(EcLDCc-CatIBs),以处理来自谷氨酸棒杆菌的含 L-赖氨酸的培养上清液。EcLDCc-CatIBs 可以很容易地在大肠杆菌中生产,然后通过简单的纯化方案,每干细胞重量可获得高达 43%的干 CatIBs。在 0.1 M 和 1 M 的 L-赖氨酸起始浓度的(重复)分批实验中,证明了 EcLDCc-CatIBs 的稳定性和可重复使用性。EcLDC-CatIBs 在反应条件下表现出很强的稳定性,估计半衰期约为 54 小时。在使用约 180-300 mg EcLDCc-CatIBs 的 30-60 ml 分批反应中,可获得 87-100%的高 DAP转化率。这导致 DAP 浓度高达 88.4 g l 和每克干 EcLDCc-CatIBs 的时空产率高达 660 g l d。因此,与文献中描述的当前最佳发酵工艺相比,这种新的 DAP 生产工艺具有竞争力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/05c4a23e7064/41598_2018_24070_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/4bfaac3acedd/41598_2018_24070_Fig1_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/4e4f7c98a3e7/41598_2018_24070_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/65f5c879fff8/41598_2018_24070_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/d257a24c721b/41598_2018_24070_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/05c4a23e7064/41598_2018_24070_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/4bfaac3acedd/41598_2018_24070_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/56818fe952e3/41598_2018_24070_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/0f42ce736e7b/41598_2018_24070_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/4e4f7c98a3e7/41598_2018_24070_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/65f5c879fff8/41598_2018_24070_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/d257a24c721b/41598_2018_24070_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ff/5895699/05c4a23e7064/41598_2018_24070_Fig7_HTML.jpg

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