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利用面包酵母在全细胞体系中提高腐胺产量以进行辅因子再生。

Improvement of cadaverine production in whole cell system with baker's yeast for cofactor regeneration.

机构信息

Department of Biological Engineering, College of Engineering, Konkuk University, Seoul, 05029, Republic of Korea.

Institute for Ubiquitous Information Technology and Applications, Konkuk University, Seoul, 05029, Republic of Korea.

出版信息

Bioprocess Biosyst Eng. 2021 Apr;44(4):891-899. doi: 10.1007/s00449-020-02497-0. Epub 2021 Jan 24.

Abstract

Cadaverine, 1,5-diaminopentane, is one of the most promising chemicals for biobased-polyamide production and it has been successfully produced up to molar concentration. Pyridoxal 5'-phosphate (PLP) is a critical cofactor for inducible lysine decarboxylase (CadA) and is required up to micromolar concentration level. Previously the regeneration of PLP in cadaverine bioconversion has been studied and salvage pathway pyridoxal kinase (PdxY) was successfully introduced; however, this system also required a continuous supply of adenosine 5'-triphosphate (ATP) for PLP regeneration from pyridoxal (PL) which add in cost. Herein, to improve the process further a method of ATP regeneration was established by applying baker's yeast with jhAY strain harboring CadA and PdxY, and demonstrated that providing a moderate amount of adenosine 5'-triphosphate (ATP) with the simple addition of baker's yeast could increase cadaverine production dramatically. After optimization of reaction conditions, such as PL, adenosine 5'-diphosphate, MgCl, and phosphate buffer, we able to achieve high production (1740 mM, 87% yield) from 2 M L-lysine. Moreover, this approach could give averaged 80.4% of cadaverine yield after three times reactions with baker's yeast and jhAY strain. It is expected that baker's yeast could be applied to other reactions requiring an ATP regeneration system.

摘要

腐胺,1,5-二氨基戊烷,是生物基聚酰胺生产最有前途的化学品之一,已经成功地生产到摩尔浓度。吡哆醛 5'-磷酸(PLP)是诱导型赖氨酸脱羧酶(CadA)的关键辅因子,需要达到微摩尔浓度水平。先前已经研究了腐胺生物转化中 PLP 的再生,并且成功引入了补救途径吡哆醛激酶(PdxY);然而,该系统还需要连续供应三磷酸腺苷(ATP),将吡哆醛(PL)转化为 PLP,这增加了成本。在此,为了进一步改进该工艺,通过应用含有 CadA 和 PdxY 的 jhAY 菌株的面包酵母建立了 ATP 再生方法,并证明通过简单添加面包酵母提供适量的三磷酸腺苷(ATP)可以显著提高腐胺的产量。在优化反应条件,如 PL、三磷酸腺苷、二磷酸腺苷、MgCl 和磷酸盐缓冲液后,我们能够从 2M L-赖氨酸中获得高产量(1740mM,87%产率)。此外,该方法在使用面包酵母和 jhAY 菌株进行三次反应后,平均可获得 80.4%的腐胺产率。预计面包酵母可应用于其他需要 ATP 再生系统的反应。

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