在连续给料批次膜反应器中以先进的酶法多克级生产核苷酸糖。

IF 4.1 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of biotechnology Pub Date : 2024-09-04 DOI:10.1016/j.jbiotec.2024.09.001
Hannes Frohnmeyer, Nikol Kodra, Lothar Elling
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引用次数: 0

摘要

酶法生产多克级核苷酸糖是一项挑战,因为只有少数几种工艺被报道用于大规模核苷酸糖生产。这些工艺主要依赖于批量合成,并且需要使用大量的酶。本研究介绍了一种利用连续给料批次膜反应器生产多克级核苷酸糖的新方法。我们成功合成了五种主要的核苷酸糖:UDP-Gal、UDP-GalNAc、UDP-GlcA、GDP-Man 和 CMP-Neu5Ac。生物催化剂的高效利用带来了高性能,包括时空产率(STY,g*L-1h-1)、总周转次数(TTN,g product per g enzyme)和适合工业相关生物工艺的高效产品形成率(g/h)。已建立的连续进料批式反应器系统在不到 15 小时的时间内连续三次生产出 8.2 克 CMP-Neu5Ac,TTN 达到 91 克产品/克酶。连续生产 UDP-GlcA 超过 28 小时,最终产品数量为 14.8 克,TTN 为 493 gP/gE。该工艺能够生产核苷酸糖并形成稳定的产物,只需极少的技术设备就能在实验室规模上生产多克量的核苷酸糖。值得注意的是,该系统具有稳健性和灵活性,可应用于各种酶法核苷酸糖合成级联。
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Advanced enzymatic multigram-scale production of nucleotide sugars in a continuous fed-batch membrane reactor

Enzymatic production of nucleotide sugars on a multigram scale presents a challenge, as only a few processes have been reported for large-scale nucleotide sugar production. They rely primarily on batch synthesis and employ exceptional amounts of enzymes. This study introduces a novel approach for the multigram-scale production of nucleotide sugars with a continuous fed-batch membrane reactor. We successfully synthesized five main nucleotide sugars: UDP-Gal, UDP-GalNAc, UDP-GlcA, GDP-Man, and CMP-Neu5Ac on a multigram scale. Efficient biocatalyst utilization results in high performance, including space-time yield (STY, g*L−1h−1), total turnover number (TTN, g product per g enzyme), and an efficient product formation rate (g/h) suitable for industrially relevant bioprocesses. The established continuous-fed batch reactor system produced up to 8.2 g CMP-Neu5Ac in three consecutive productions in less than 15 h with satisfying TTNs of 91 gProduct/gEnzyme. Continuous production of UDP-GlcA over 28 h resulted in a final product amount of 14.8 g and TTN of 493 gP/gE. This process enables the production of nucleotide sugars with stable product formation, requiring minimal technical equipment for multigram quantities of nucleotide sugars at the laboratory scale. Notably, the system exhibited robustness and flexibility, allowing its application to various enzymatic nucleotide sugar synthesis cascades.

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来源期刊
Journal of biotechnology
Journal of biotechnology 工程技术-生物工程与应用微生物
CiteScore
8.90
自引率
2.40%
发文量
190
审稿时长
45 days
期刊介绍: The Journal of Biotechnology has an open access mirror journal, the Journal of Biotechnology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal provides a medium for the rapid publication of both full-length articles and short communications on novel and innovative aspects of biotechnology. The Journal will accept papers ranging from genetic or molecular biological positions to those covering biochemical, chemical or bioprocess engineering aspects as well as computer application of new software concepts, provided that in each case the material is directly relevant to biotechnological systems. Papers presenting information of a multidisciplinary nature that would not be suitable for publication in a journal devoted to a single discipline, are particularly welcome.
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