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在乳球菌中构建木糖同化途径,以利用玉米秸秆原料进行高产 d-乳酸发酵。

Constructing xylose-assimilating pathways in Pediococcus acidilactici for high titer d-lactic acid fermentation from corn stover feedstock.

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China.

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China.

出版信息

Bioresour Technol. 2017 Dec;245(Pt B):1369-1376. doi: 10.1016/j.biortech.2017.05.128. Epub 2017 May 22.

Abstract

Xylose-assimilating pathway was constructed in a d-lactic acid producing Pediococcus acidilactici strain and evolutionary adapted to yield a co-fermentation strain P. acidilactici ZY15 with 97.3g/L of d-lactic acid and xylose conversion of 92.6% obtained in the high solids content simultaneous saccharification and co-fermentation (SSCF) of dry dilute acid pretreated and biodetoxified corn stover feedstock. The heterologous genes encoding xylose isomerase (xylA) and xylulokinase (xylB) were screened and integrated into the P. acidilactici chromosome. The metabolic flux to acetic acid in phosphoketolase pathway was re-directed to pentose phosphate pathway by substituting the endogenous phosphoketolase gene (pkt) with the heterologous transketolase (tkt) and transaldolase (tal) genes. The xylose-assimilating ability of the newly constructed P. acidilactici strain was significantly improved by adaptive evolution. This study provided an important strain and process prototype for high titer d-lactic acid production from lignocellulose feedstock with efficient xylose assimilation.

摘要

木糖同化途径在产 d-乳酸的戊糖片球菌菌株中构建,并经过进化适应,以产生共发酵菌株 P. acidilactici ZY15,在高固体含量的同步糖化和共发酵(SSCF)中,该菌株可从干燥稀酸预处理和生物解毒的玉米秸秆原料中获得 97.3g/L 的 d-乳酸和 92.6%的木糖转化率。筛选并整合了编码木糖异构酶(xylA)和木酮糖激酶(xylB)的异源基因到戊糖片球菌染色体中。通过用异源转酮醇酶(tkt)和转醛醇酶(tal)基因替代内源性磷酸酮醇酶基因(pkt),将磷酸酮解酶途径中的代谢通量重新导向戊糖磷酸途径。通过适应性进化,新构建的戊糖片球菌菌株的木糖同化能力得到了显著提高。本研究为高效利用木质纤维素原料生产高浓度 d-乳酸提供了重要的菌株和工艺原型。

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