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

• -methyltransferases, or engineered lepidopteran mevalonate pathways. The substrate promiscuity of TSs not only expands the structural diversity of terpenes but also highlights their potential for the discovery of novel terpenoids via combinatorial biosynthesis. In this review, we focus on the current knowledge on

• multisubstrate terpene synthases (MSTSs) and highlight their potential applications. Keywords: noncanonical terpene; substrate promiscuity; synthetic biology; terpene synthase; terpenoid; Introduction Terpenoids constitute the largest class of natural products with more than 80000 known structures [1] and a

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

• positions of the A ring of drimenol by heterologous expression and gene knockout. Furthermore, we tested the substrate promiscuity of CavA with drimenol analogs. CavA exhibited the ability to accept albicanol (5) and drim-8-ene-11-ol (6) as substrates, leading to the formation of the hydroxylated

• to note that CavA exhibits limited substrate promiscuity, predominantly targeting the drimenol skeleton with minor variations. This selectivity may be attributed to the structural configuration of the enzyme, which appears to be finely tuned to recognize and interact with specific features of the

Research progress on the pharmacological activity, biosynthetic pathways, and biosynthesis of crocins

• Zhongwei Hua,
• Nan Liu and
• Xiaohui Yan

Beilstein J. Org. Chem. 2024, 20, 741–752, doi:10.3762/bjoc.20.68

• , and UGTs are the key players in crocin biosynthesis. Empowered by the genomic, transcriptomic, and metabolomic analysis and biochemical characterization, many such enzymes have been identified. Some proteins, exemplified by GjCCD4a and GjUGT94E13, exhibit broad substrate promiscuity and high catalytic

Chemoenzymatic synthesis of macrocyclic peptides and polyketides via thioesterase-catalyzed macrocyclization

• Senze Qiao,
• Zhongyu Cheng and
• Fuzhuo Li

Beilstein J. Org. Chem. 2024, 20, 721–733, doi:10.3762/bjoc.20.66

• macrocyclizations (Scheme 1c). NAC thioester and other related mimics (such as coenzyme A (CoA), phosphopantetheine, and thiophenol) span the gap between the chemical synthesis and biosynthesis languages and expand the substrate promiscuity of TE domains. This bridge makes the in vitro TE-catalyzed macrocyclization

Synthesis of C6-modified mannose 1-phosphates and evaluation of derived sugar nucleotides against GDP-mannose dehydrogenase

• Sanaz Ahmadipour,
• Alice J. C. Wahart,
• Jonathan P. Dolan,
• Laura Beswick,
• Chris S. Hawes,
• Robert A. Field and
• Gavin J. Miller

Beilstein J. Org. Chem. 2022, 18, 1379–1384, doi:10.3762/bjoc.18.142

• ). Conclusion We have developed a synthetic approach to a small series of C6-modified mannose 1-phosphates (6-amino, 6-chloro and 6-thio) and with these tools further explored the substrate promiscuity of the mannose pyrophosphorylase from S. enterica, observing that larger (than canonical OH) chlorine is

Cytochrome P450 monooxygenase-mediated tailoring of triterpenoids and steroids in plants

• Karan Malhotra and
• Jakob Franke

Beilstein J. Org. Chem. 2022, 18, 1289–1310, doi:10.3762/bjoc.18.135

• classes in plants, we expect that this number will rise quickly in years to come. Several of our examples highlight the substrate promiscuity embedded within ancient CYP families, which enables rapid functional extension to acquire unique catalytic functions during duplication events [15][26][79]. The

• . polyphylla and TfCYP90B50-TfCYP82J17 in T. foenum-graecum resulted in the highest diosgenin (13) production. Diosgenin (13) biosynthesis in distantly related plants is an example of catalytic plasticity embedded within the ancient CYP90Bs. Especially CYPs from large families often show high substrate

• promiscuity which facilitates duplication events resulting in neofunctionalisation [15]. Ellarinacin (15) is a defence-related arborinane-type triterpenoid that was recently discovered in bread wheat (Triticum aestivum) by genome mining (Figure 6B) [26]. The ellarinacin gene cluster encodes the three CYP

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

• selectivity observed from the transformation of their natural substrate. For the exploration of substrate promiscuity of plant-derived ent-kaurene oxidases, twenty combinations of type II/I diterpene TCs were coexpressed with one of the two ent-kaurene oxidases (CYPs) and their native CPRs from different

• are shown in Figure 10a) [129]. While only a limited number of bacterial CYPs for terpene modification has been characterized, many of them were reported to exhibit substrate promiscuity (Supporting Information File 1, Table S2). These promiscuous CYPs do not necessarily co-localize with TC-encoding

• -isozizaene (33) synthase mutants that produce different sesquiterpene skeletons. Substrate promiscuity and engineering of CYPs. a) Selected examples from using a CYP library to oxidize various monoterpenes. b) Rational engineering of P450BM3 for epoxidation of amorphadiene (21). F87A/A328L and R47L/Y51F

Characterization of non-heme iron aliphatic halogenase WelO5* from Hapalosiphon welwitschii IC-52-3: Identification of a minimal protein sequence motif that confers enzymatic chlorination specificity in the biosynthesis of welwitindolelinones

• Qin Zhu and
• Xinyu Liu

Beilstein J. Org. Chem. 2017, 13, 1168–1173, doi:10.3762/bjoc.13.115

• , this study, along with the recent characterization of the AmbO5 protein, collectively confirmed the presence of a signature sequence motif in the C-terminus of this newly discovered halogenase enzyme family that confers substrate promiscuity and specificity. These observations may guide the rational

• chimera revealed that a C-terminal sequence motif plays a role in the substrate tolerance and provided insights into the origin of substrate promiscuity in this family of proteins [18]. In this work, we report the characterization of the third WelO5-type protein, WelO5*, for the biogenesis 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

• 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

• performed in vivo substrate promiscuity tests following a combinatorial approach [41][66]. The resulting products entailed pimarane- and abietane-type diterpenes as well as the trans-clerodane type diterpene kolavenol, a putative intermediate in the salvinorin A biosynthesis. Other bifunctional diterpene

Carbohydrate PEGylation, an approach to improve pharmacological potency

• M. Eugenia Giorgi,
• Rosalía Agusti and
• Rosa M. de Lederkremer

Beilstein J. Org. Chem. 2014, 10, 1433–1444, doi:10.3762/bjoc.10.147

• PEGylation of a glycoprotein can be performed in three steps. First, the sialic acid is removed from the native protein with a sialidase and subsequently Sia-PEG is transfered to the uncovered terminal Gal units of the linked glycan taking advantage of the substrate promiscuity of the sialyltransferase