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利用谷氨酸棒杆菌在一罐式工艺中从木糖中微需氧生长解耦生产α-酮戊二酸和琥珀酸。

Microaerobic growth-decoupled production of α-ketoglutarate and succinate from xylose in a one-pot process using Corynebacterium glutamicum.

机构信息

Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Jülich, Germany.

Bioeconomy Science Center (BioSC), Forschungszentrum Jülich GmbH, Jülich, Germany.

出版信息

Biotechnol J. 2021 Sep;16(9):e2100043. doi: 10.1002/biot.202100043. Epub 2021 Jun 18.

Abstract

BACKGROUND

Lignocellulosic biomass is the most abundant raw material on earth. Its efficient use for novel bio-based materials is essential for an emerging bioeconomy. Possible building blocks for such materials are the key TCA-cycle intermediates α-ketoglutarate and succinate. These organic acids have a wide range of potential applications, particularly in use as monomers for established or novel biopolymers. Recently, Corynebacterium glutamicum was successfully engineered and evolved towards an improved utilization of d-xylose via the Weimberg pathway, yielding the strain WMB2 . The Weimberg pathway enables a carbon-efficient C5-to-C5 conversion of d-xylose to α-ketoglutarate and a shortcut route to succinate as co-product in a one-pot process.

METHODS AND RESULTS

C. glutamicum WMB2 was grown under dynamic microaerobic conditions on d-xylose, leading to the formation of comparably high amounts of succinate and only small amounts of α-ketoglutarate. Subsequent carbon isotope labeling experiments verified the targeted production route for both products in C. glutamicum WMB2 . Fed-batch process development was initiated and the effect of oxygen supply and feeding strategy for a growth-decoupled co-production of α-ketoglutarate and succinate were studied in detail. The finally established fed-batch production process resulted in the formation of 78.4 mmol L (11.45 g L ) α-ketoglutarate and 96.2 mmol L (11.36 g L ) succinate.

CONCLUSION

The developed one-pot process represents a promising approach for the combined supply of bio-based α-ketoglutarate and succinate. Future work will focus on tailor-made down-stream processing of both organic acids from the fermentation broth to enable their application as building blocks in chemical syntheses. Alternatively, direct conversion of one or both acids via whole-cell or cell-free enzymatic approaches can be envisioned; thus, extending the network of value chains starting from cheap and renewable d-xylose.

摘要

背景

木质纤维素生物质是地球上最丰富的原料。高效利用这种新型生物基材料对于新兴的生物经济至关重要。此类材料的可能构建块是关键的三羧酸 (TCA) 循环中间体α-酮戊二酸和琥珀酸。这些有机酸具有广泛的潜在应用,特别是作为现有或新型生物聚合物的单体。最近,通过 Weimberg 途径,成功地对谷氨酸棒杆菌进行了工程改造和进化,以提高 d-木糖的利用率,得到了菌株 WMB2。Weimberg 途径能够以碳效率高的方式将 d-木糖转化为α-酮戊二酸,并在一锅法中作为副产物生成琥珀酸的捷径。

方法和结果

在动态微氧条件下,C. glutamicum WMB2 以 d-木糖为碳源生长,导致琥珀酸和少量α-酮戊二酸的形成量相当高。随后的碳同位素标记实验验证了 C. glutamicum WMB2 中这两种产物的靶向生产途径。启动了分批补料工艺的开发,并详细研究了供氧和进料策略对生长解偶联的α-酮戊二酸和琥珀酸联产的影响。最终建立的分批补料生产工艺形成了 78.4 mmol L(11.45 g L)α-酮戊二酸和 96.2 mmol L(11.36 g L)琥珀酸。

结论

所开发的一锅法代表了一种有前途的方法,可用于同时供应生物基α-酮戊二酸和琥珀酸。未来的工作将集中在从发酵液中对这两种有机酸进行定制化的下游加工,以使其能够作为化学合成中的构建块应用。或者,可以设想通过全细胞或无细胞酶法直接转化一种或两种酸,从而扩展从廉价可再生 d-木糖开始的价值链网络。

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