Enhanced limonene production in a fast-growing cyanobacterium through combinatorial metabolic engineering

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Po-Cheng Lin , Fuzhong Zhang , Himadri B. Pakrasi
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引用次数: 36

Abstract

Terpenoids are a large and diverse group of natural products with commercial applications. Microbial production of terpenes is considered as a feasible approach for the stable supply of these complex hydrocarbons. Cyanobacteria, photosynthetic prokaryotes, are attractive hosts for sustainable bioproduction, because these autotrophs require only light and CO2 for growth. Despite cyanobacteria having been engineered to produce a variety of compounds, their productivities of terpenes are generally low. Further research is needed to determine the bottleneck reactions for enhancing terpene production in cyanobacteria. In this study, we engineered the fast-growing cyanobacterium Synechococcus elongatus UTEX 2973 to produce a commercially-used terpenoid, limonene. We identified a beneficial mutation in the gene encoding geranylgeranyl pyrophosphate synthase crtE, leading to a 2.5-fold increase in limonene production. The engineered strain produced 16.4 ​mg ​L−1 of limonene at a rate of 8.2 ​mg ​L−1 day−1, which is 8-fold higher than limonene productivities previously reported in other cyanobacterial species. Furthermore, we employed a combinatorial metabolic engineering approach to optimize genes involved in the upstream pathway of limonene biosynthesis. By modulating the expression of genes encoding the enzymes in the MEP pathway and the geranyl pyrophosphate synthase, we showed that optimization of the expression level is critical to enhance limonene production in cyanobacteria.

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通过组合代谢工程提高快速生长蓝藻的柠檬烯产量
萜类化合物是一大类具有商业用途的天然产物。微生物生产萜烯被认为是稳定供应这些复杂碳氢化合物的可行途径。蓝藻,光合原核生物,是可持续生物生产的有吸引力的宿主,因为这些自养生物只需要光和二氧化碳来生长。尽管蓝藻经过改造可以产生多种化合物,但它们的萜烯产量通常很低。需要进一步的研究来确定提高蓝藻中萜烯产量的瓶颈反应。在这项研究中,我们设计了快速生长的蓝藻长聚球菌UTEX 2973,以生产一种商业用途的萜类化合物柠檬烯。我们在编码香叶基焦磷酸合成酶crtE的基因中发现了一个有益的突变,导致柠檬烯产量增加2.5倍。该工程菌株以8.2 mg L−1 day−1的速率产生16.4 mg L−1柠檬烯,比以前报道的其他蓝藻物种的柠檬烯产量高8倍。此外,我们采用组合代谢工程的方法来优化参与柠檬烯生物合成上游途径的基因。通过调节MEP通路和香叶基焦磷酸合成酶编码基因的表达,我们发现优化表达水平对提高蓝藻柠檬烯的产量至关重要。
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来源期刊
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
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