Institute of Biochemistry, Heinrich-Heine University Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany.
Microb Cell Fact. 2021 Sep 20;20(1):183. doi: 10.1186/s12934-021-01673-5.
The aryltetralin lignan (-)-podophyllotoxin is a potent antiviral and anti-neoplastic compound that is mainly found in Podophyllum plant species. Over the years, the commercial demand for this compound rose notably because of the high clinical importance of its semi-synthetic chemotherapeutic derivatives etoposide and teniposide. To satisfy this demand, (-)-podophyllotoxin is conventionally isolated from the roots and rhizomes of Sinopodophyllum hexandrum, which can only grow in few regions and is now endangered by overexploitation and environmental damage. For these reasons, targeting the biosynthesis of (-)-podophyllotoxin precursors or analogues is fundamental for the development of novel, more sustainable supply routes.
We recently established a four-step multi-enzyme cascade to convert (+)-pinoresinol into (-)-matairesinol in E. coli. Herein, a five-step multi-enzyme biotransformation of (-)-matairesinol to (-)-deoxypodophyllotoxin was proven effective with 98 % yield at a concentration of 78 mg/L. Furthermore, the extension of this cascade to a sixth step leading to (-)-epipodophyllotoxin was evaluated. To this end, seven enzymes were combined in the reconstituted pathway involving inter alia three plant cytochrome P450 monooxygenases, with two of them being functionally expressed in E. coli for the first time.
Both, (-)-deoxypodophyllotoxin and (-)-epipodophyllotoxin, are direct precursors to etoposide and teniposide. Thus, the reconstitution of biosynthetic reactions of Sinopodophyllum hexandrum as an effective multi-enzyme cascade in E. coli represents a solid step forward towards a more sustainable production of these essential pharmaceuticals.
芳基四氢萘木脂素（-）-鬼臼毒素是一种有效的抗病毒和抗肿瘤化合物，主要存在于鬼臼属植物中。多年来，由于其半合成化学治疗衍生物依托泊苷和替尼泊苷的临床重要性，对该化合物的商业需求显著增加。为了满足这一需求，（-）-鬼臼毒素通常从 Sinopodophyllum hexandrum 的根和根茎中分离出来，而 Sinopodophyllum hexandrum 只能在少数地区生长，现在正受到过度开发和环境破坏的威胁。出于这些原因，针对（-）-鬼臼毒素前体或类似物的生物合成是开发新型、更可持续供应途径的基础。
我们最近在大肠杆菌中建立了一个四步多酶级联反应，将（+）-松脂素转化为（-）-马替瑞林。在此，通过五步多酶生物转化，以 78mg/L 的浓度将（-）-马替瑞林有效转化为（-）-脱氧鬼臼毒素，产率为 98%。此外，还评估了将该级联反应扩展到第六步以生成（-）-表鬼臼毒素的效果。为此，在涉及三种植物细胞色素 P450 单加氧酶的重建途径中组合了七种酶，其中两种酶首次在大肠杆菌中功能性表达。
（-）-脱氧鬼臼毒素和（-）-表鬼臼毒素都是依托泊苷和替尼泊苷的直接前体。因此，将 Sinopodophyllum hexandrum 的生物合成反应在大肠杆菌中有效地重建为多酶级联反应，是朝着更可持续地生产这些重要药物迈出的坚实一步。
