Discovery and characterization of diterpene synthases in Chamaecyparis formosensis Matsum. which participated in an unprecedented diterpenoid biosynthesis route in conifer

AbstractAnalyzing the gene sequences of terpene synthase (TPS) may contribute to a better understanding of terpenes biosynthesis and evolution of phylogenetic taxonomy.Chamaecyparis formosensisis an endemic and precious conifer of Taiwan. To understand the biosynthesis mechanism of terpenes in this tree, a full length of putative mono-TPS, named asCf-Pin(GeneBank accession no. EU099434), was obtained by PCR method and RACE extension. TheCf-Pinhas an 1887-bp open reading frame and encodes 628 amino acids. To identify the function ofCf-Pin,the recombinant protein fromEscherichia coliwas incubated with geranyl diphosphate, produced one major product, the structure of which was elucidated. GC/MS analysis and matching of retention time and mass spectrum with authentic standards revealed that this product isα-pinene. This is the first report of cloning of a mono-TPS and functionally expressed inE. coliand which could be identified asα-pinene synthase from a Cupressaceae conifer.

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Characterization of CYP76AH4 clarifies phenolic diterpenoid biosynthesis in the Lamiaceae

Organic & Biomolecular Chemistry ◽

10.1039/c3ob41885e ◽

2013 ◽

Vol 11 (44) ◽

pp. 7650 ◽

Cited By ~ 51

Author(s):

Jiachen Zi ◽

Reuben J. Peters

Keyword(s):

Diterpenoid Biosynthesis

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Platensimycin and Platencin Biosynthesis in Streptomyces platensis, Showcasing Discovery and Characterization of Novel Bacterial Diterpene Synthases

Natural Product Biosynthesis by Microorganisms and Plants, Part A - Methods in Enzymology ◽

10.1016/b978-0-12-394290-6.00008-2 ◽

2012 ◽

pp. 163-186 ◽

Cited By ~ 4

Author(s):

Michael J. Smanski ◽

Ryan M. Peterson ◽

Ben Shen

Keyword(s):

Streptomyces Platensis ◽

Diterpene Synthases

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Biochemical characterization of diterpene synthases of Taiwania cryptomerioides expands the known functional space of specialized diterpene metabolism in gymnosperms

The Plant Journal ◽

10.1111/tpj.14513 ◽

2019 ◽

Vol 100 (6) ◽

pp. 1254-1272 ◽

Cited By ~ 2

Author(s):

Li‐Ting Ma ◽

Yi‐Ru Lee ◽

Nai‐Wen Tsao ◽

Sheng‐Yang Wang ◽

Philipp Zerbe ◽

...

Keyword(s):

Biochemical Characterization ◽

Functional Space ◽

Taiwania Cryptomerioides ◽

Diterpene Synthases

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Isolation and characterization of β-cadinene synthase cDNA from Chamaecyparis formosensis Matsum

Holzforschung ◽

10.1515/hf-2011-0224 ◽

2012 ◽

Vol 66 (5) ◽

pp. 569-576 ◽

Cited By ~ 3

Author(s):

Pei-Min Kuo ◽

Keng-Hao Hsu ◽

Yi-Ru Lee ◽

Fang-Hua Chu ◽

Sheng-Yang Wang

Keyword(s):

Functional Expression ◽

Major Product ◽

Gas Chromatography Mass Spectrometry ◽

Synthesis Mechanism ◽

E Coli ◽

Isolation And Characterization ◽

Ms Analysis ◽

Chamaecyparis Formosensis ◽

Rapid Amplification

Abstract Chamaecyparis formosensis is a precious conifer endemic in Taiwan. To understand the sesquiterpene synthesis mechanism in this tree, full-length cDNA of a putative sesquiterpene synthase (sesqui-TPS), designated Cf-Cad, was obtained by rapid amplification of complementary DNA ends-polymerase chain reaction. Cf-Cad is 1812 bp in length. To identify its function, recombinant protein from Escherichia coli was incubated with farnesyl diphosphate, which produced one major product, the structure of which was elucidated by gas chromatography/mass spectrometry (GC/MS) analysis. GC/MS analysis, GC retention time and MS matching with authentic standards revealed that the major product was β-cadinene. This is the first report of the cloning, functional expression in E. coli and identification of a sesqui-TPS from a Cupressaceae conifer.

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