Six enzymes from mayapple that complete the biosynthetic pathway to the etoposide aglycone

被引:355
作者
Lau, Warren [1 ]
Sattely, Elizabeth S. [1 ]
机构
[1] Stanford Univ, Dept Chem Engn, Stanford, CA 94305 USA
关键词
PODOPHYLLUM-HEXANDRUM; ARYLTETRALIN LIGNANS; CLONING; PURIFICATION; EXPRESSION; REDUCTASE; PROTEIN; YEAST;
D O I
10.1126/science.aac7202
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Podophyllotoxin is the natural product precursor of the chemotherapeutic etoposide, yet only part of its biosynthetic pathway is known. We used transcriptome mining in Podophyllum hexandrum (mayapple) to identify biosynthetic genes in the podophyllotoxin pathway. We selected 29 candidate genes to combinatorially express in Nicotiana benthamiana (tobacco) and identified six pathway enzymes, including an oxoglutarate-dependent dioxygenase that closes the core cyclohexane ring of the aryltetralin scaffold. By coexpressing 10 genes in tobacco-these 6 plus 4 previously discovered-we reconstitute the pathway to (-)-4'-desmethylepipodophyllotoxin (the etoposide aglycone), a naturally occurring lignan that is the immediate precursor of etoposide and, unlike podophyllotoxin, a potent topoisomerase inhibitor. Our results enable production of the etoposide aglycone in tobacco and circumvent the need for cultivation of mayapple and semisynthetic epimerization and demethylation of podophyllotoxin.
引用
收藏
页码:1224 / 1228
页数:7
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