Modular pathway engineering for the microbial production of branched-chain fatty alcohols.
Modular pathway engineering for the microbial production of branched-chain fatty alcohols.
复制标题
DOI：
10.1186/s13068-017-0936-4
复制
发表时间：
2017
影响因子：
6.3
通讯作者：
Zhang F
中科院分区：
文献类型：
--
作者：
Jiang W;Qiao JB;Bentley GJ;Liu D;Zhang F
与直链脂肪醇相比，支链长链脂肪醇(BLFL)在C12-C18范围内的固有结构特性使其更适合作为柴油替代品和其他工业应用。虽然微生物已经实现了直链长链脂肪醇的生产，但生物合成BLFL的报道从未见过。在这项工作中，我们在大肠杆菌中设计了四种不同的生物合成途径来生产BLFL。然后，我们采用模块化工程方法优化了α-酮酸前体的供应，生产出了高选择性的奇链和偶链BLFL，分别达到总脂肪醇的70%和75%。酰基-ACP和酒精生产模块也进行了广泛的优化，以平衡酶表达水平和比率，从而使BLFL滴度提高了6.5倍。表现最好的菌株在补料分批发酵罐中过表达了6个工程操纵子的14个基因，产量为350 mg/L。模块化工程策略成功地促进了BLFL的微生物生产，并使我们能够快速优化具有高滴度和产品特异性的新BLFL途径。更广泛地说，这项工作为以高滴度和高产量生产BLFL和与BLFL相关的工业相关化学品提供了途径和知识。本文的在线版本(doi：10.1186/s13068-0170936-4)包含补充材料，可供授权用户使用。
The intrinsic structural properties of branched long-chain fatty alcohols (BLFLs) in the range of C12 to C18 make them more suitable as diesel fuel replacements and for other industrial applications than their straight-chain counterparts. While microbial production of straight long-chain fatty alcohols has been achieved, biosynthesis of BLFLs has never been reported. In this work, we engineered four different biosynthetic pathways in Escherichia coli to produce BLFLs. We then employed a modular engineering approach to optimize the supply of α-keto acid precursors and produced either odd-chain or even-chain BLFLs with high selectivity, reaching 70 and 75% of total fatty alcohols, respectively. The acyl-ACP and alcohol-producing modules were also extensively optimized to balance enzyme expression level and ratio, resulting in a 6.5-fold improvement in BLFL titers. The best performing strain overexpressed 14 genes from 6 engineered operons and produced 350 mg/L of BLFLs in fed-batch fermenter. The modular engineering strategy successfully facilitated microbial production of BLFLs and allowed us to quickly optimize new BLFL pathway with high titers and product specificity. More generally, this work provides pathways and knowledge for the production of BLFLs and BLFL-related, industry-relevant chemicals in high titers and yields. The online version of this article (doi:10.1186/s13068-017-0936-4) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
3.7
作者：
Engler, Carola;Kandzia, Romy;Marillonnet, Sylvestre
通讯作者：
Marillonnet, Sylvestre
DOI：
10.1126/science.1191652
发表时间：
2010-10-01
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Ajikumar PK;Xiao WH;Tyo KE;Wang Y;Simeon F;Leonard E;Mucha O;Phon TH;Pfeifer B;Stephanopoulos G
通讯作者：
Stephanopoulos G
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：
8.4
作者：
Garg, Shivani;Rizhsky, Ludmila;Nikolau, Basil J.
通讯作者：
Nikolau, Basil J.
影响因子：
4.8
作者：
Lai, CY;Cronan, JE
通讯作者：
Cronan, JE