Nepeta cataria L. (catnip) can serve as a chassis for the engineering of secondary metabolic pathways.

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology Letters Pub Date : 2024-10-01 Epub Date: 2024-05-08 DOI:10.1007/s10529-024-03489-w

Marcus Geissler, Christoph Neubauer, Yuriy V Sheludko, Adrian Brückner, Heribert Warzecha

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Abstract

Objective: Evaluation of Nepeta cataria as a host with specific endogenous metabolite background for transient expression and metabolic engineering of secondary biosynthetic sequences.

Results: The reporter gene gfp::licBM3 as well as three biosynthetic genes leading to the formation of the cannabinoid precursor olivetolic acid were adopted to the modular cloning standard GoldenBraid, transiently expressed in two chemotypes of N. cataria and compared to Nicotiana benthamiana. To estimate the expression efficiency in both hosts, quantification of the reporter activity was carried out with a sensitive and specific lichenase assay. While N. benthamiana exhibited lichenase activity of 676 ± 94 μmol g^-1 s^-1, N. cataria cultivar '1000', and the cultivar 'Citriodora' showed an activity of 37 ± 8 μmol g^-1 s^-1 and 18 ± 4 μmol g^-1 s^-1, respectively. Further, combinatorial expression of genes involved in cannabinoid biosynthetic pathway acyl-activating enzyme 1 (aae1), olivetol synthase (ols) and olivetolic acid cyclase (oac) in N. cataria cv. resulted presumably in the in vivo production of olivetolic acid glycosides.

Conclusion: Nepeta cataria is amenable to Agrobacterium-mediated transient expression and could serve as a novel chassis for the engineering of secondary metabolic pathways and transient evaluation of heterologous genes.

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猫薄荷（Nepeta cataria L.）可以作为次级代谢途径工程的底盘。

目的结果：将报告基因 gfp::licBM3 和导致大麻素前体橄榄醇酸形成的三个生物合成基因采用模块化克隆方法进行克隆：结果：报告基因 gfp::licBM3 以及导致大麻素前体橄榄醇酸形成的三个生物合成基因被采用到模块化克隆标准 GoldenBraid 中，在两种化学型的 N. cataria 中瞬时表达，并与烟草本根进行比较。为了估算两种宿主的表达效率，采用了一种灵敏而特异的地衣酶测定法对报告活性进行量化。N. benthamiana 的地衣酶活性为 676 ± 94 μmol g-1 s-1，而 N. cataria 栽培品种 "1000 "和栽培品种 "Citriodora "的活性分别为 37 ± 8 μmol g-1 s-1 和 18 ± 4 μmol g-1 s-1。此外，在 N. cataria cv.中组合表达涉及大麻素生物合成途径的酰基活化酶 1（aae1）、橄榄醇合成酶（ols）和橄榄醇酸环化酶（oac）的基因，可能会导致体内产生橄榄醇酸苷：结论：Nepeta cataria 适合农杆菌介导的瞬时表达，可作为次生代谢途径工程和异源基因瞬时评估的新型底盘。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.