Enhancing structural diversity of terpenoids by multisubstrate terpene synthases

• Min Li and
• Hui Tao

Beilstein J. Org. Chem. 2024, 20, 959–972, doi:10.3762/bjoc.20.86

• substrates, MSTSs can also accept partially cyclized substrates. A class I diterpene synthase SiTPS8 from Setaria italicais is capable of utilizing three copalyl pyrophosphate (CPP) stereoisomers that were generated by different class II TSs, including ent-CPP (19), (+)-CPP (20), and syn-CPP (21), to

Functional characterisation of twelve terpene synthases from actinobacteria

• Anuj K. Chhalodia,
• Houchao Xu,
• Georges B. Tabekoueng,
• Binbin Gu,
• Kizerbo A. Taizoumbe,
• Lukas Lauterbach and
• Jeroen S. Dickschat

Beilstein J. Org. Chem. 2023, 19, 1386–1398, doi:10.3762/bjoc.19.100

• phylogenetically related enzymes were in one case not expressed and in two cases inactive, suggesting pseudogenisation in the respective branch of the phylogenetic tree. Furthermore, a diterpene synthase for allokutznerene and a sesterterpene synthase for sesterviolene were identified. Keywords: actinomycetes

• inactive enzymes were obtained and one enzyme was not expressed. Another interesting discovery was the identification of a diterpene synthase from Kutzneria kofuensis that selectively produces allokutznerene. This compound was previously only known as a side product from a closely related phomopsene

Characterization of a new fusicoccane-type diterpene synthase and an associated P450 enzyme

• Jia-Hua Huang,
• Jian-Ming Lv,
• Liang-Yan Xiao,
• Qian Xu,
• Fu-Long Lin,
• Gao-Qian Wang,
• Guo-Dong Chen,
• Sheng-Ying Qin,
• Dan Hu and
• Hao Gao

Beilstein J. Org. Chem. 2022, 18, 1396–1402, doi:10.3762/bjoc.18.144

• nature and possess a variety of biological activities. Up to date, only five fusicoccane-type diterpene synthases have been identified. Here, we identify a two-gene biosynthetic gene cluster containing a new fusicoccane-type diterpene synthase gene tadA and an associated cytochrome P450 gene tadB from

• . Keywords: cytochrome P450 enzyme; diterpene synthase; gene cluster; genome mining; site-directed mutagenesis; Introduction Terpenoids are a large class of natural products that attract extensive attention, due to not only their potential applications in pharmaceuticals, agrochemicals, etc. but also due to

Efficient production of clerodane and ent-kaurane diterpenes through truncated artificial pathways in Escherichia coli

• Fang-Ru Li,
• Xiaoxu Lin,
• Qian Yang,
• Ning-Hua Tan and
• Liao-Bin Dong

Beilstein J. Org. Chem. 2022, 18, 881–888, doi:10.3762/bjoc.18.89

• ), have attracted great attention from chemists and biologists due to their intriguing chemical structures and broad pharmacological functions [1][2][3][4]. The vast structural diversity of diterpenoids arise biosynthetically from the following two stages: i) diterpene synthase (DTS, also called diterpene

Targeting active site residues and structural anchoring positions in terpene synthases

• Anwei Hou and
• Jeroen S. Dickschat

Beilstein J. Org. Chem. 2021, 17, 2441–2449, doi:10.3762/bjoc.17.161

• , turning SmTS1 from a sesterterpene into a diterpene synthase. This article gives rational explanations for these findings that may generally allow for protein engineering of other terpene synthases to improve their catalytic efficiency or to change their functions. Keywords: biosynthesis; enzyme

• ). SmTS1 has a low amino acid sequence identity to other characterised TPSs, with the diterpene synthase (DTS) for cattleyene from Streptomyces cattleya as one of the closest relatives, which shows only 29% sequence identity [17]. We have recently shown that the sum of the calculated van der Waals volumina

• C3 of the substrate (G182 in Figure 1) and assists in substrate ionisation. The introduction of steric bulk at this position blocked this movement for SdS, resulting in inactivity [11]. Diterpene synthase activity of SmTS1 variants While the A222V enzyme variant did not convert GFPP, presumably

Current understanding and biotechnological application of the bacterial diterpene synthase CotB2

• Ronja Driller,
• Daniel Garbe,
• Norbert Mehlmer,
• Monika Fuchs,
• Keren Raz,
• Dan Thomas Major,
• Thomas Brück and
• Bernhard Loll

Beilstein J. Org. Chem. 2019, 15, 2355–2368, doi:10.3762/bjoc.15.228

• date, CotB2 represents the best studied bacterial diterpene synthase. Its reaction mechanism has been addressed by isoptope labeling, targeted mutagenesis and theoretical computations in the gas phase, as well as full enzyme molecular dynamic simulations. By X-ray crystallography different snapshots of

• three Mg2+ ions [1][22]. Notably, the PtmT1 diterpene synthase from S. platensis lacks both catalytic motifs [20][23]. The NSE/DTE motif illustrates the distribution of DTE motifs, which are predominantly found in plant TPSs, and NSE motifs, which are more common in bacterial and fungal TPSs. For the

• , fungicidal and tumorstatic effects [32]. A key player in the biosynthesis of cyclooctatin 5 is the bacterial diterpene synthase CotB2. Different research teams have investigated CotB2 by means of biochemical [30][31][38], biophysical [33][34][35], structural biology [36][37][38][39] and computational

Volatiles from three genome sequenced fungi from the genus Aspergillus

• Jeroen S. Dickschat,
• Ersin Celik and
• Nelson L. Brock

Beilstein J. Org. Chem. 2018, 14, 900–910, doi:10.3762/bjoc.14.77

• enzyme XP_001276070 (ACLA_076850), likely a bifunctional diterpene synthase (DTS), seems to be not expressed in laboratory culture, but another function cannot be excluded for this enzyme. Esters were the predominant class of compounds emitted by A. clavatus. The observed pattern of volatiles was very

18-Hydroxydolabella-3,7-diene synthase – a diterpene synthase from Chitinophaga pinensis

• Jeroen S. Dickschat,
• Jan Rinkel,
• Patrick Rabe,
• Arman Beyraghdar Kashkooli and
• Harro J. Bouwmeester

Beilstein J. Org. Chem. 2017, 13, 1770–1780, doi:10.3762/bjoc.13.171

• Wageningen, The Netherlands, Swammerdam Institute for Life Sciences, University of Amsterdam, Sciencepark 904, 1098 XH Amsterdam, The Netherlands 10.3762/bjoc.13.171 Abstract The product obtained in vitro from a diterpene synthase encoded in the genome of the bacterium Chitinophaga pinensis, an enzyme

• reaction were addressed by isotopic labelling experiments. Heterologous expression of the diterpene synthase in Nicotiana benthamiana resulted in the production of 18-hydroxydolabella-3,7-diene also in planta, while the results from the heterologous expression in E. coli were shown to be reproducible

• based on the identification of this compound and its Cope rearrangement product β-elemene (2) formed by the thermal impact during GC–MS analysis [19] in E. coli headspace extracts under heterologous expression of the terpene synthase gene (Scheme 1). Here we present the diterpene synthase activity of

Opportunities and challenges for the sustainable production of structurally complex diterpenoids in recombinant microbial systems

• Katarina Kemper,
• Max Hirte,
• Markus Reinbold,
• Monika Fuchs and
• Thomas Brück

Beilstein J. Org. Chem. 2017, 13, 845–854, doi:10.3762/bjoc.13.85

• the rapid identification of (di)terpenes. Andersen-Ranberg and co-workers reported recently on the creation of a synthetic collection of monofunctional Class I/Class II diterpene synthase combinations, which lead to high stereoselective syntheses of an impressive number of previously unknown or

• unamenable diterpenes with labdane- and clerodane-type structures [66]. Additional findings were provided by Jia and co-workers [67], who demonstrated high substrate promiscuity of a plant and a fungal Class I diterpene synthase. This study involved general substrates of diterpene cyclases like GGPP and its

• example of these bifunctional enzymes was published by Chen and coworkers [60], who managed to crystalize catalytic domains of PaFS, a diterpene synthase from Phomopsis amygdali. The formation of GGPP is located in a C-terminal α-domain with very low sequence identity to the N-terminal Class I terpene