Additional diterpenes from Physcomitrella patens synthesized by copalyl diphosphate/kaurene synthase (PpCPS/KS)

IF 6.1 2区生物学 Q1 PLANT SCIENCES

Plant Physiology and Biochemistry Pub Date : 2015-11-01 DOI:10.1016/j.plaphy.2015.07.011

Xin Zhan, Søren Spanner Bach, Nikolaj Lervad Hansen, Christina Lunde, Henrik Toft Simonsen

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引用次数: 20

Abstract

The bifunctional diterpene synthase, copalyl diphosphate/kaurene synthase from the moss Physcomitrella patens (PpCPS/KS), catalyses the formation of at least four diterpenes, including ent-beyerene, ent-sandaracopimaradiene, ent-kaur-16-ene, and 16-hydroxy-ent-kaurene. The enzymatic activity has been confirmed through generation of a targeted PpCPS/KS knock-out mutant in P. patens via homologous recombination, through transient expression of PpCPS/KS in Nicotiana benthamiana, and expression of PpCPS/KS in E. coli. GC-MS analysis of the knock-out mutant shows that it lacks the diterpenoids, supporting that all are products of PpCPS/KS as observed in N. benthamiana and E. coli. These results provide additional knowledge of the mechanism of this bifunctional diterpene synthase, and are in line with proposed reaction mechanisms in kaurene biosynthesis.

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二磷酸共酚/丁香烯合成酶(PpCPS/KS)合成小笠藻中附加二萜

双功能二萜合成酶，即来自小立藓(physcomitellla patens, PpCPS/KS)的共聚二磷酸/丁香烯合成酶，催化至少四种二萜的形成，包括对-对苯二烯、对-sandaracopimaradiene、对-丁香烯-16烯和16-羟基对-丁香烯。通过同源重组在patens中产生PpCPS/KS靶向敲除突变体，在Nicotiana benthamiana中瞬时表达PpCPS/KS，以及在大肠杆菌中表达PpCPS/KS，证实了该酶的活性。GC-MS分析表明，敲除突变体缺乏二萜，支持在benthamiana和E. coli中观察到的PpCPS/KS产物。这些结果为这种双功能二萜合成酶的机制提供了额外的知识，并且与提出的karen生物合成反应机制一致。

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来源期刊

Plant Physiology and Biochemistry 生物-植物科学

CiteScore

11.10

自引率

3.10%

发文量

410

审稿时长

33 days

期刊介绍： Plant Physiology and Biochemistry publishes original theoretical, experimental and technical contributions in the various fields of plant physiology (biochemistry, physiology, structure, genetics, plant-microbe interactions, etc.) at diverse levels of integration (molecular, subcellular, cellular, organ, whole plant, environmental). Opinions expressed in the journal are the sole responsibility of the authors and publication does not imply the editors'' agreement. Manuscripts describing molecular-genetic and/or gene expression data that are not integrated with biochemical analysis and/or actual measurements of plant physiological processes are not suitable for PPB. Also "Omics" studies (transcriptomics, proteomics, metabolomics, etc.) reporting descriptive analysis without an element of functional validation assays, will not be considered. Similarly, applied agronomic or phytochemical studies that generate no new, fundamental insights in plant physiological and/or biochemical processes are not suitable for publication in PPB. Plant Physiology and Biochemistry publishes several types of articles: Reviews, Papers and Short Papers. Articles for Reviews are either invited by the editor or proposed by the authors for the editor''s prior agreement. Reviews should not exceed 40 typewritten pages and Short Papers no more than approximately 8 typewritten pages. The fundamental character of Plant Physiology and Biochemistry remains that of a journal for original results.