Metabolic engineering of Pseudomonas sp. strain VLB120 as platform biocatalyst for the production of isobutyric acid and other secondary metabolites.
Metabolic engineering of Pseudomonas sp. strain VLB120 as platform biocatalyst for the production of isobutyric acid and other secondary metabolites.
复制标题
假单胞菌的代谢工程。菌株VLB120作为平台生物催化剂，用于产生异丁酸和其他次级代谢产物。
DOI：
10.1186/1475-2859-13-2
复制
发表时间：
2014-01-07
影响因子：
6.4
通讯作者：
Schmid A
中科院分区：
文献类型：
--
作者：
Lang K;Zierow J;Buehler K;Schmid A
近年来，埃利希途径衍生的化学物质的产生在多种宿主中显示，如大肠杆菌、谷氨酸棒杆菌和酵母。例如，在大肠杆菌中以显著的滴度和产率证明了异丁酸的产生。然而，这些实施例由于各自生物体的发酵生长模式而遭受副产物形成。我们的目标是使用假单胞菌属菌株VLB120（一种专性好氧生物）作为宿主菌株，建立一种新的好氧底盘来合成异丁酸和其他有趣的代谢物。乳酸乳球菌（Lactococcus lactis）2-酮酸脱羧酶基因kivd在Ps.菌株VLB120能够通过缬氨酸合成途径（埃利希途径）生产异丁酸和异丁醇。这表明存在催化异丁醛还原和氧化的染色体编码的醇和醛脱氢酶。此外，我们表明，该菌株具有一个完整的异丁酸代谢途径，通过异丁酰辅酶A进入缬氨酸降解的化合物。解决了三个关键问题以允许和优化异丁酸合成：i）通过宿主内在酶使异丁酸降解最小化，ii）构建合适的表达系统和iii）中心碳代谢的流线化，最终导致产生高达26.8 ± 1.5mM的异丁酸，碳产率为0.12 ± 0.01g gglc-1。使用定制的表达系统增加流向异丁酸的通量和防止前体和产物降解的组合允许在Ps中有效生产异丁酸。sp.菌株VLB120。这将是为这种散装化学品开发连续反应工艺的基础。
Over the recent years the production of Ehrlich pathway derived chemicals was shown in a variety of hosts such as Escherichia coli, Corynebacterium glutamicum, and yeast. Exemplarily the production of isobutyric acid was demonstrated in Escherichia coli with remarkable titers and yields. However, these examples suffer from byproduct formation due to the fermentative growth mode of the respective organism. We aim at establishing a new aerobic, chassis for the synthesis of isobutyric acid and other interesting metabolites using Pseudomonas sp. strain VLB120, an obligate aerobe organism, as host strain. The overexpression of kivd, coding for a 2-ketoacid decarboxylase from Lactococcus lactis in Ps. sp. strain VLB120 enabled for the production of isobutyric acid and isobutanol via the valine synthesis route (Ehrlich pathway). This indicates the existence of chromosomally encoded alcohol and aldehyde dehydrogenases catalyzing the reduction and oxidation of isobutyraldehyde. In addition we showed that the strain possesses a complete pathway for isobutyric acid metabolization, channeling the compound via isobutyryl-CoA into valine degradation. Three key issues were addressed to allow and optimize isobutyric acid synthesis: i) minimizing isobutyric acid degradation by host intrinsic enzymes, ii) construction of suitable expression systems and iii) streamlining of central carbon metabolism finally leading to production of up to 26.8 ± 1.5 mM isobutyric acid with a carbon yield of 0.12 ± 0.01 g gglc-1. The combination of an increased flux towards isobutyric acid using a tailor-made expression system and the prevention of precursor and product degradation allowed efficient production of isobutyric acid in Ps. sp. strain VLB120. This will be the basis for the development of a continuous reaction process for this bulk chemicals.
登录
查看更多内容
影响因子：
4.4
作者：
Connor, Michael R.;Liao, James C.
通讯作者：
Liao, James C.
影响因子：
5.4
作者：
Blank, Lars M.;Ionidis, Georgios;Schmid, Andreas
通讯作者：
Schmid, Andreas
影响因子：
5
作者：
Hüsken, LE;Beeftink, R;Wery, J
通讯作者：
Wery, J
影响因子：
3.8
作者：
Gross, Rainer;Buehler, Katja;Schmid, Andreas
通讯作者：
Schmid, Andreas
影响因子：
48
作者：
Choi, KH;Gaynor, JB;Schweizer, HP
通讯作者：
Schweizer, HP