Chemo-enzymatic total synthesis: current approaches toward the integration of chemical and enzymatic transformations

• Ryo Tanifuji and
• Hiroki Oguri

Beilstein J. Org. Chem. 2024, 20, 1693–1712, doi:10.3762/bjoc.20.151

• prenyltransferase (PT) domain located at the C-terminus of BscA (Scheme 2A). Subsequently, the terpene cyclase (TC) domain at the N-terminal of BscA generates fusicocca-2,10(14)-diene (6), which bears the common 5/8/5 fused tricyclic scaffold common to this natural products family. Sequential oxidative conversions

Discovery and biosynthesis of bacterial drimane-type sesquiterpenoids from Streptomyces clavuligerus

• Dongxu Zhang,
• Wenyu Du,
• Xingming Pan,
• Xiaoxu Lin,
• Fang-Ru Li,
• Qingling Wang,
• Qian Yang,
• Hui-Min Xu and
• Liao-Bin Dong

Beilstein J. Org. Chem. 2024, 20, 815–822, doi:10.3762/bjoc.20.73

• calidoustene C, DrtB from Aspergillus calidoustus functions as a dual-functional enzyme, comprising two domains: a HAD-like hydrolase domain fused with a terpene cyclase domain. Initially, FPP is cyclized into the drimenyl diphosphate in a class II terpene cyclase manner, which is then processed by the

• utilizing the EFI-genome neighborhood tool (EFI-GNT), sequence alignment, and manual BLAST analysis [33]. Leveraging our previous discovery of SsDMS from Streptomyces showdoensis, we identified a homologous terpene cyclase (sclav_p0068) in S. clavuligerus. This enzyme shares a 50% sequence similarity with

• ]. Inspection of the BGC identified additional three P450s. Thus, the entire cluster mainly encompasses a DMS (CavC), a Nudix hydrolase (CavB), a class I terpene cyclase (CavF), and three P450s (CavA, CavE, and CavG), and it was designated as the cav cluster (Figure 3a and Table S2 in Supporting Information

Genome mining of labdane-related diterpenoids: Discovery of the two-enzyme pathway leading to (−)-sandaracopimaradiene in the fungus Arthrinium sacchari

• Fumito Sato,
• Terutaka Sonohara,
• Shunta Fujiki,
• Akihiro Sugawara,
• Yohei Morishita,
• Taro Ozaki and
• Teigo Asai

Beilstein J. Org. Chem. 2024, 20, 714–720, doi:10.3762/bjoc.20.65

• of TCs in fungi. Keywords: diterpenoids; fungi; genome mining; labdane; terpene cyclase; Introduction Terpenoids are a structurally diverse family of natural products, including more than 80,000 compounds [1]. In the biosynthesis of terpenoids, terpene cyclases (TCs) add structural diversity and

Production of non-natural 5-methylorsellinate-derived meroterpenoids in Aspergillus oryzae

• Jia Tang,
• Yixiang Zhang and
• Yudai Matsuda

Beilstein J. Org. Chem. 2024, 20, 638–644, doi:10.3762/bjoc.20.56

• -MOA as a substrate for dearomatizing farnesylation. Subsequently, we introduced five terpene cyclase genes involved in DMOA-derived meroterpenoid biosynthesis, namely, adrI, trt1, ausL, insA7, and insB2, individually into the A. oryzae transformant that already expresses the four genes constructed

Unraveling the role of prenyl side-chain interactions in stabilizing the secondary carbocation in the biosynthesis of variexenol B

• Moe Nakano,
• Rintaro Gemma and
• Hajime Sato

Beilstein J. Org. Chem. 2023, 19, 1503–1510, doi:10.3762/bjoc.19.107

• as the C–H–π interaction between the carbocation intermediate and the Phe residue of terpene cyclase in the biosynthesis of sesterfisherol [21], and the intricated rearrangement reaction mechanism promoted by the equilibrium state of the homoallyl cation and the cyclopropylcarbinyl cation in the

Strategies to access the [5-8] bicyclic core encountered in the sesquiterpene, diterpene and sesterterpene series

• Cécile Alleman,
• Charlène Gadais,
• Laurent Legentil and
• François-Hugues Porée

Beilstein J. Org. Chem. 2023, 19, 245–281, doi:10.3762/bjoc.19.23

Anti-inflammatory aromadendrane- and cadinane-type sesquiterpenoids from the South China Sea sponge Acanthella cavernosa

• Shou-Mao Shen,
• Qing Yang,
• Yi Zang,
• Jia Li,
• Xueting Liu and
• Yue-Wei Guo

Beilstein J. Org. Chem. 2022, 18, 916–925, doi:10.3762/bjoc.18.91

• shifts led to aristolane-type carbocation intermediate G, which was further deprotonated to afford 9-aristolene (H) [29]. Multiple-step oxidation on H furnished the structure of 3. The terpene cyclase catalyzed the cyclization of cadinene-type sesquiterpenes using FPP as the substrate, which is first

Understanding the role of active site residues in CotB2 catalysis using a cluster model

• Keren Raz,
• Ronja Driller,
• Thomas Brück,
• Bernhard Loll and
• Dan T. Major

Beilstein J. Org. Chem. 2020, 16, 50–59, doi:10.3762/bjoc.16.7

• , we compared the energy profiles of the terpene cyclase CotB2 reaction obtained in the gas phase and using an active site model. The calculations used identical QM methods, facilitating a direct comparison. We presented evidence for the important role played by the active site residues in CotB2 on the

Bacterial terpene biosynthesis: challenges and opportunities for pathway engineering

• Eric J. N. Helfrich,
• Geng-Min Lin,
• Christopher A. Voigt and
• Jon Clardy

Beilstein J. Org. Chem. 2019, 15, 2889–2906, doi:10.3762/bjoc.15.283

• . Keywords: bacterial sesquiterpenes and diterpenes; cytochrome P450; pathway engineering; synthetic biology; terpene biosynthesis; terpene cyclase; Introduction Evolutionary diversification of terpene biosynthetic pathways has resulted in the largest and most structurally diverse class of specialized

• a linear polyene with branching methyl groups that form the core hydrocarbon structure in a single enzyme-catalyzed step [9]. The enzyme, which is called terpene cyclase, holds the linear methyl-branched polyene in a defined conformation that initiates a series of carbocation-driven cyclizations and

• placed on terpene pathways from bacteria, as their biosynthetic pathways usually have the genes encoding the terpene cyclase and modifying enzyme in close proximity, which simplifies both analysis and pathway engineering. The review will begin with a brief description of terpene families with a special

Harnessing enzyme plasticity for the synthesis of oxygenated sesquiterpenoids

• Melodi Demiray,
• David J. Miller and
• Rudolf K. Allemann

Beilstein J. Org. Chem. 2019, 15, 2184–2190, doi:10.3762/bjoc.15.215

• -FDPs to sesquiterpenoids that may have applications in healthcare and agriculture. These results inform us of both the utility and limitations that non-natural functional groups have upon terpene cyclase-catalysed reaction cascades supporting the design of future biocatalytic syntheses. In particular

Bipolenins K–N: New sesquiterpenoids from the fungal plant pathogen Bipolaris sorokiniana

• Chin-Soon Phan,
• Hang Li,
• Simon Kessler,
• Peter S. Solomon,
• Andrew M. Piggott and
• Yit-Heng Chooi

Beilstein J. Org. Chem. 2019, 15, 2020–2028, doi:10.3762/bjoc.15.198

• biosynthetic gene cluster (tpc) for terpestacin (12) has been recently identified from Bipolaris maydis [36]. A didomain sesterterpene synthase (tpcA) with a terpene cyclase domain and polyprenyltransferase domain was demonstrated to be responsible for the production of the sesterterpene backbone of 12. A

Inherent atomic mobility changes in carbocation intermediates during the sesterterpene cyclization cascade

• Hajime Sato,
• Takaaki Mitsuhashi,
• Mami Yamazaki,
• Ikuro Abe and
• Masanobu Uchiyama

Beilstein J. Org. Chem. 2019, 15, 1890–1897, doi:10.3762/bjoc.15.184

• these two methyl groups are critical for the preorganization of GFPP in the biosynthetic pathways leading to sesterfisherol and quiannulatene. Keywords: biosynthesis; carbocation; DFT; substrate recognition; terpene cyclase; Introduction Terpene synthases are thought to have four main roles: (i

• terpene cyclase active site [3][5][6]. Although many terpene cyclases are known [6][7][8][9][10], it is still challenging to identify the precise initial conformation of the oligoprenyl diphosphate substrate in the active site, even by X-ray crystal structure determination. This is because the substrate

• can sometimes bind to the active site in an unreactive conformation [11]. Recently, Siegel and Tantillo reported an innovative method for predicting the docking mode of carbocation intermediates in terpene cyclase [12][13], based on QM calculation and computational docking with the Rosetta protein

Stereochemical investigations on the biosynthesis of achiral (Z)-γ-bisabolene in Cryptosporangium arvum

• Jan Rinkel and
• Jeroen S. Dickschat

Beilstein J. Org. Chem. 2019, 15, 789–794, doi:10.3762/bjoc.15.75

• enantiomers of NPP, which is surprising in the light of the fact that these reactive tertiary allylic diphosphates were often found to result in a complex mixture of terpene cyclase products and Mg2+-catalysed spontaneous hydrolysis products for other TSs [29][30]. While the reaction with (R)-NPP leads to

Herpetopanone, a diterpene from Herpetosiphon aurantiacus discovered by isotope labeling

• Xinli Pan,
• Nicole Domin,
• Sebastian Schieferdecker,
• Hirokazu Kage,
• Martin Roth and
• Markus Nett

Beilstein J. Org. Chem. 2017, 13, 2458–2465, doi:10.3762/bjoc.13.242

• Biology, Hans Knöll Institute, Beutenbergstr. 11a, 07745 Jena, Germany 10.3762/bjoc.13.242 Abstract The genome of the predatory bacterium Herpetosiphon aurantiacus 114-95T harbors a number of biosynthesis genes, including four terpene cyclase genes. To identify the terpenes biosynthesized from H

• numerous bacterial terpene cyclase genes [2][3]. Both in vitro and in vivo approaches involving recombinant enzymes are commonly pursued for their functional characterization [4]. Care must be taken, however, in interpreting the results of these analyses, as the products of terpene cyclases are often

• subject to enzymatic modifications in their native producers. The exclusive testing of a terpene cyclase might hence unveil a biosynthetic intermediate rather than the final product of a secondary metabolite pathway [5][6]. To avoid this problem, we here describe a method for the identification 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

• diversity of terpene products is obtained by precise modulation of cyclization and rearrangement steps performed by terpene cyclase enzymes [31], initial functional groups are introduced by hydroxylation of the carbon backbone with highly specific P450 monooxygenases [42][43][44]. At present, terpene

Mechanistic investigations on six bacterial terpene cyclases

• Patrick Rabe,
• Thomas Schmitz and
• Jeroen S. Dickschat

Beilstein J. Org. Chem. 2016, 12, 1839–1850, doi:10.3762/bjoc.12.173

• terpene cyclases from bacterial genomes. Altogether, a number of ca. 1000 terpene cyclase genes are found in the genomes of sequenced bacteria [10], and about 50 bacterial terpene cyclases have so far been characterised for their products [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27

• bacterial [28][35] and fungal [36][37] terpene cyclases. Results and Discussion Incubation of a recombinant terpene cyclase from Streptomyces viridochromogenes DSM 40736 (NCBI accession number WP_039931950) with farnesyl diphosphate (FPP) yielded a single product that was identified as α-amorphene (1

• established the structure of T-muurolol (2). The absolute configuration was determined as (1R,6S,7R,10R)-(+)-T-muurolol (2) from its optical rotary power ([α]D23 = +99.4 (c 1.10, CH2Cl2)). This is the same compound as was reported from a terpene cyclase from Streptomyces clavuligerus (accession number

The EIMS fragmentation mechanisms of the sesquiterpenes corvol ethers A and B, epi-cubebol and isodauc-8-en-11-ol

• Patrick Rabe and
• Jeroen S. Dickschat

Beilstein J. Org. Chem. 2016, 12, 1380–1394, doi:10.3762/bjoc.12.132

• carry a labelling (>99% 13C) in specific positions that can easily be located, if the cyclisation mechanism of the terpene cyclase is known. Furthermore, 13C NMR spectroscopy can be used to experimentally locate the labelling if the cyclisation mechanism is unidentified. As we have shown in two previous

• [16][17]. In the present study we have used the same approach to investigate the fragmentation mechanisms for corvol ethers A and B, two sesquiterpene ethers with unique carbon skeletons that are made by a terpene cyclase from Kitasatospora setae [18], and the sesquiterpene alcohols epi-cubebol and