Trans-4-hydroxy-L-proline production by the cyanobacterium Synechocystis sp. PCC 6803

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Fabian Brandenburg , Eleni Theodosiou , Carolin Bertelmann, Marcel Grund, Stephan Klähn, Andreas Schmid, Jens O. Krömer
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引用次数: 7

Abstract

Cyanobacteria play an important role in photobiotechnology. Yet, one of their key central metabolic pathways, the tricarboxylic acid (TCA) cycle, has a unique architecture compared to most heterotrophs and still remains largely unexploited. The conversion of 2-oxoglutarate to succinate via succinyl-CoA is absent but is by-passed by several other reactions. Overall, fluxes under photoautotrophic growth conditions through the TCA cycle are low, which has implications for the production of chemicals. In this study, we investigate the capacity of the TCA cycle of Synechocystis sp PCC 6803 for the production of trans-4-hydroxy-L-proline (Hyp), a valuable chiral building block for the pharmaceutical and cosmetic industries. For the first time, photoautotrophic Hyp production was achieved in a cyanobacterium expressing the gene for the L-proline-4-hydroxylase (P4H) from Dactylosporangium sp. strain RH1. Interestingly, while elevated intracellular Hyp concentrations could be detected in the recombinant Synechocystis strains under all tested conditions, detectable Hyp secretion into the medium was only observed when the pH of the medium exceeded 9.5 and mostly in the late phases of the cultivation. We compared the rates obtained for autotrophic Hyp production with published sugar-based production rates in E. coli. The land-use efficiency (space-time yield) of the phototrophic process is already in the same order of magnitude as the heterotrophic process considering sugar farming as well. But, the remarkable plasticity of the cyanobacterial TCA cycle promises the potential for a 23–55 fold increase in space-time yield when using Synechocystis. Altogether, these findings contribute to a better understanding of bioproduction from the TCA cycle in photoautotrophs and broaden the spectrum of chemicals produced in metabolically engineered cyanobacteria.

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聚囊蓝细菌pcc6803生产反式4-羟基- l -脯氨酸的研究
蓝藻在光生物技术中发挥着重要作用。然而,与大多数异养生物相比,它们的关键中心代谢途径之一三羧酸(TCA)循环具有独特的结构,并且在很大程度上仍未被开发。没有通过琥珀酰辅酶a将2-氧戊二酸转化为琥珀酸盐,但有几个其他反应绕过。总的来说,在光自养生长条件下通过三羧酸循环的通量很低,这对化学品的生产有影响。在这项研究中,我们研究了聚囊藻sp PCC 6803的TCA循环生产反式4-羟基- l -脯氨酸(Hyp)的能力,这是一种有价值的手性成分,用于制药和化妆品工业。首次在表达Dactylosporangium sp.菌株RH1的l -脯氨酸-4-羟化酶(P4H)基因的蓝藻中实现了光自养hypp的产生。有趣的是,虽然在所有测试条件下,重组聚胞菌菌株的细胞内Hyp浓度都有所升高,但只有当培养基的pH超过9.5时,培养基中才有可检测到的Hyp分泌,而且大部分是在培养后期。我们比较了在大肠杆菌中获得的自养产氢酶的速率和已发表的糖基产氢酶的速率。考虑到食糖种植,光养过程的土地利用效率(时空产量)已与异养过程处于同一数量级。但是,蓝藻TCA循环的显著可塑性保证了使用聚囊藻时时空产量增加23-55倍的潜力。总之,这些发现有助于更好地理解光自养生物中TCA循环的生物生产,并拓宽了代谢工程蓝藻产生的化学物质的范围。
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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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