Engineered production of isoprene from the model green microalga Chlamydomonas reinhardtii

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic Engineering Communications Pub Date : 2023-06-01 DOI:10.1016/j.mec.2023.e00221
Razan Z. Yahya, Gordon B. Wellman, Sebastian Overmans, Kyle J. Lauersen
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Abstract

Isoprene is a clear, colorless, volatile 5-carbon hydrocarbon that is one monomer of all cellular isoprenoids and a platform chemical with multiple applications in industry. Many plants have evolved isoprene synthases (IspSs) with the capacity to liberate isoprene from dimethylallyl diphosphate (DMADP) as part of cellular thermotolerance mechanisms. Isoprene is hydrophobic and volatile, rapidly leaves plant tissues and is one of the main carbon emission sources from vegetation globally. The universality of isoprenoid metabolism allows volatile isoprene production from microbes expressing heterologous IspSs. Here, we compared heterologous overexpression from the nuclear genome and localization into the plastid of four plant terpene synthases (TPs) in the green microalga Chlamydomonas reinhardtii. Using sealed vial mixotrophic cultivation, direct quantification of isoprene production was achieved from the headspace of living cultures, with the highest isoprene production observed in algae expressing the Ipomoea batatas IspS. Perturbations of the downstream carotenoid pathway through keto carotenoid biosynthesis enhanced isoprene titers, which could be further enhanced by increasing flux towards DMADP through heterologous co-expression of a yeast isopentenyl-DP delta isomerase. Multiplexed controlled-environment testing revealed that cultivation temperature, rather than illumination intensity, was the main factor affecting isoprene yield from the engineered alga. This is the first report of heterologous isoprene production from a eukaryotic alga and sets a foundation for further exploration of carbon conversion to this commodity chemical.

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利用莱茵衣藻工程化生产异戊二烯
异戊二烯是一种透明、无色、挥发性的5碳碳氢化合物,是所有细胞类异戊二烯中的一种单体,也是一种在工业中具有多种应用的平台化学品。作为细胞耐热机制的一部分,许多植物已经进化出具有从二甲基烯丙基二磷酸(DMADP)中释放异戊二烯的能力的异戊二烯合成酶(IspSs)。异戊二烯具有疏水性和挥发性,能迅速离开植物组织,是全球植被的主要碳排放源之一。类异戊二烯代谢的普遍性允许表达异源IspSs的微生物产生挥发性异戊二烯。在这里,我们比较了来自核基因组的异源过表达和绿色微藻莱茵衣藻中四种植物萜烯合成酶(TP)在质体中的定位。使用密封小瓶混合营养培养,从活培养物的顶部空间实现了异戊二烯生产的直接定量,在表达Ipomoea batatas IspS的藻类中观察到最高的异戊二烯生产。通过酮类胡萝卜素生物合成对下游类胡萝卜素途径的干扰增强了异戊二烯滴度,通过异源共表达酵母异戊烯基DPδ异构酶增加向DMADP的流量可以进一步增强异戊二烯滴度。多重控制环境试验表明,影响工程藻类异戊二烯产量的主要因素是培养温度,而不是光照强度。这是第一份由真核藻类生产异源异戊二烯的报告,为进一步探索碳转化为这种商品化学品奠定了基础。
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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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