Enzymatic Cascade RX
Enzymatic Cascade RX
Enzymatic Cascade RX
Reviews Chemie
Keywords:
electrophilic cascades ·
natural products ·
nucleophilic cascades ·
pericyclic cascades ·
radical cascades
Angewandte
Chemie
&&&& 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 1. Examples of natural product chemical syntheses exemplifying the five mechanistic categories of cascade reactions. A) A nucleophilic
cyclization cascade in the total synthesis of tetronasin by Ley and colleagues.[7] B) An electrophilic cascade involving an epoxy-olefin cyclization in
the total synthesis of hemibrevetoxin B by Holton and colleagues.[8] C) A radical cyclization cascade in the total synthesis of morphine by Parker
and Fokas.[9] D) A pericyclic cascades involving Diels–Alder and [3+2] cycloadditions in the total synthesis of vindorosine by Boger and
colleagues.[10] E) Transition-metal-catalyzed cascades involving multiple ring-opening/ring-closing olefin-metatheses in the total synthesis of
cyanthiwigin U by Pfeiffer and Phillips.[11] AIBN = 2,2’-azobisisobutyronitrile, KHMDS = potassium bis(trimethylsilyl)amide, MOM = methoxymethyl,
N-PSP = N-(phenylseleno)phthalimide, TIPB = triisopropylbenzene, Ts = p-tolylsulphonyl.
controlled chemical transformations in living organisms. In an remnants of which exist today, for example, in the ribonu-
RNA-world scenario, they were preceded by catalytic RNAs, cleoprotein ribosomes that carry out protein biosynthesis.[12]
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
There are some cascade reactions in primary metabolism, the nonribosomal peptide/polyketide natural product frame-
interconversion of small-molecule metabolites into product- works. A large subset of polyketide synthases (PKSs) and
(s) carried out by all cells. For example, the tandem action of essentially all of the nonribosomal peptide synthetases
thiolase and hydroxymethylglutaryl-CoA synthase converts (NRPSs) hold onto the growing intermediate chains as
three acetyl-CoA molecules to the C6 half thioester HMG- covalent thioesters and thus qualify as cascade catalysts.[13]
CoA.[13] However, the major enzymatic complexity-generat- Carbanion equivalents as nucleophiles. The celebrated
ing events occur in conditional pathways, known collectively deoxyerythronolide B (DEB) synthase, for example, takes
as secondary metabolism. seven molecules of 2S-methylmalonyl-CoA and elongates
This is the realm of natural product biosynthesis, that is, them to the 14-carbon linear thioester and then carries out
biosynthesis of the isoprenoids, alkaloids, phenylpropanoids, regiospecific cyclization to release the 14-member DEB
polyketides, and nonribosomal peptides that account for most macrocyclic lactone as only one of 1024 possible diastereo-
of the approximately 500,000 known natural products.[13] mers[17] (Scheme 2 A). Analogously, in the biosynthesis of
These also include the bioactive molecular frameworks that tetracycline, the enzyme trio of OxyABC builds a tethered 19-
have been the targets of hundreds of natural product synthetic carbon nonaketidyl thioester chain (Scheme 2 B). Tailoring
studies by chemists over the past 80 years. enzymes then carry out transannular aldol and Claisen
Cascade reaction planning, implementation, and analysis condensation reactions to release preteramid as the first
have been core activities of synthetic chemists in the context soluble intermediate.[18]
of biomimetic or “bioinspired” routes.[3a, 6] Many of these To build these nucleophilic chain-elongating cascade
synthetic campaigns were undertaken before the enzymes of reactions, polyketide synthases need carbon nucleophiles to
a particular pathway were identified, purified, and examined generate the C C bonds. These are typically the CoA
for mechanism and specificity. Nonetheless, they often had thioester enolates of malonyl and methylmalonyl acids (all
predictive power, given that chemical mechanisms play out in seven monomers incorporated in DEB above). The thioester
biology. Indeed, a recent review by Liu and co-workers has grouping activates C1 as the electrophile and adjacent C2 as
catalogued examples of dozens of organic named reactions in the nucleophile for iterative thioclaisen condensations (see
enzymatic catalysis,[14] many of which are key steps in both Ref. [13] for a review).
nonenzymatic and enzymatic synthetic cascades. Amines as nucleophiles. In contrast to polyketide cas-
In this Review, we delve into enzymatic examples of cades, where C C bond formations are the chain elongation
cascade reactions to illustrate how barriers for multistep steps, NRPSs make amide bonds and use the amino nitrogen
transformations are lowered in specific enzyme active sites. atoms of tethered amino acid monomers as the chain-
These are part and parcel of the ability of enzymes involved in elongating nucleophiles.[19] In the active sites of the conden-
natural product biosynthesis to build complexity from simple sation/chain elongation catalytic domains of NRPS assembly
primary metabolites. The coverage is not meant to be lines, the predominant -NH3+ ionization states must be
encyclopedic but rather to illustrate how some of the organic converted into nucleophilic NH2 groups (Scheme 3 A). Con-
named reactions are put to use in cascades within a given comitantly, in hybrid NRPS/PKS assembly lines, amines and
enzyme active site, natures equivalent of the one-pot thioester enolates are the requisite nucleophiles, for example,
synthetic reaction. We will mention a few tandem reactions, in rapamycin (Scheme 3 B) or FK506 assembly.[20]
involving the consecutive action of two enzymes to promul- An example of an unleashed cascade reaction occurs in
gate the cascades. We exclude multienzyme participations in the action of the fungal trimodular NRPS that makes
full biosynthetic pathway reconstitutions, although we have fumiquinazoline F, in which anthranilate, tryptophan, and
noted elsewhere the remarkable efficiency of enzymes to alanine building blocks are combined into a fused tricyclic
function in short pathways that build remarkable scaffold quinazoline core.[21] The assembly line enzyme builds a teth-
complexity.[13, 15] ered linear anthranilyl-Trp-Ala-thioester, which undergoes
While one could divide the presentation according to intramolecular capture by the anthranilyl amine, releasing
specific natural product categories, we think it more useful to a presumptive 6,10-macrocycle. This is never detected.
categorize enzymatic cascade reactions by mechanism. To Instead, one observes only the transannular, cyclodehydrated
that end we use the Nicolaou[3a] approach of nucleophilic, quinazoline product (Scheme 4).
electrophilic, radical-based, and pericyclic cascades. There is We note in the section on radical-driven cascades below
no enzymatic analogue of nonenzymatic olefin metathesis that the tethered heptapeptidyl chain in the vancomycin
cascades and no indication that palladium or platinum are synthetase assembly line is acted on successively by three
involved in biology. Molybdenum is used in a small number of “thwarted oxygenases” that make side-chain radicals that
enzymes[16] but not detectably for olefin metatheses. form cross links rather than undergo hydroxylations, as
emblematic of tailoring reactions that occur on NRPS
assembly lines. There are many examples of tailoring reaction
3. Enzymatic Nucleophilic Cascades on Assembly on PKS assembly lines as well, including Michael additions by
Lines side chain -OH groups on conjugated enoyl thioesters to form
cyclic ethers (Scheme 5).[22]
Cascade reactions initiated or carried out by nucleophiles
are common in the enzymatic assembly lines that build many
thousands of polyketide, nonribosomal peptide, and hybrid
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 2. Two polyketides generated by enzymatic cascades. A) The 14-member deoxyerythronolide B (DEB) lactone is released from the three-
subunit DEB synthase assembly line after incorporation of seven methylmalonyl units. B) Oxytetracycline biosynthesis involves a chain elongation
cascade to a nineteen carbon nonaketonyl thioester that is converted into the tetracyclic pretetramide product.
3.1. Nucleophilic Enzymatic Cascades Enabled by Redox Steps adduct and then reductively releases it through hydride
transfer from the co-substrate NADH to yield the nascent
There are a number of cases where enzymes that carry out linear peptide aldehyde (Scheme 6). This can be cyclized
an oxidative or reductive change on a substrate/nascent through attack of the N-terminal Cys1 amino group on the
product uncover reactions that can be categorized as cascade Val7 aldehyde to give the cyclic imine. The equilibrium in
reactions. One simple case, extending the above discussion of favor of cyclization is further driven by addition of the Cys1
NRPS assembly lines, is the recent discovery of the pathway side-chain -SH onto the imine to yield the cyclic thiazolidine.
to the cyclic peptide lugdunin, produced by Staphylococcus This is the accumulating form of lugdunin, which acts as an
lugdunensis, which competes with pathogenic Staphylococcus antibiotic against S. aureus strains in the oral cavity.
aureus strains in human oral cavities.[23] The lugdunin NRPS Another cascade reaction set in motion by a redox step
assembly line generates a tethered heptapeptidyl-enzyme with a nicotinamide co-substrate is catalyzed by the S-
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Scheme 3. Nonribosomal peptide synthetase assembly lines carry out chain-elongation cascades to the heptapeptide vancomycin (A) and the
hybrid NRPS/PKS product rapamycin (B), in which amine nucleophiles are utilized for the insertion of amino acids.
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 4. The fumiquinazoline F NRPS assembly line carries out a cascade employing the amino group of the anthranilyl-1 residue for cyclizing
release, followed by transannular capture of the transient 6,0-macrocycle to yield the tricyclic quinazolinone framework.
adenosylhomocysteine (SAH) lyase, the key enzyme in phile to drive subsequent carbocycle formation. Examples of
returning homocysteine carbons to the cellular metabolic this enzymatic strategy include the formation of plant iridoid
pool after SAM has been used for millions of methyl transfers monoterpene scaffolds[25] and also the tricyclic core of
in every cell cycle (Scheme 7). The enzyme is erroneously bacterial polycyclic tetramate macrolactams.[26] Iridoid scaf-
termed a hydrolase because the thioether linkage in SAH is folds, such as in nepetalactone and the secologanin-derived
converted into free homocysteine and adenosine. The thio- moiety in the oncology agent vincristine, arise from geranyl-
ether is stable to hydrolysis. Instead the SAH lyase first diphosphate (GPP) that is processed to 8-oxogeranial (1,8-
oxidizes C-3 of the ribose ring to the ketone while generating geranyl dialdehyde). Hydride conjugate addition from
NADH, which is kept tightly bound in the active site.[24] The NADPH by iridoid synthase sets up the enolate that reacts
value of the alcohol to ketone oxidation is in acidification of intramolecularly through a Michael cyclization to form the
the adjacent C4 OH since the resultant carbanion is now irodial nucleus nepetalactol (Scheme 8 A).[25b] In analogy, the
stabilizable as the enolate anion. This easily accessible ikarugamycin biosynthetic enzyme IkaB similarly transfers
carbanion can be used to eliminate the homocysteine a hydride to the terminal olefin of the nascent product from
moiety with C5 S cleavage to yield the conjugated enone the IkaA polyketide synthase (Scheme 8 B). A proposed 4+2
with a 4,5-exomethylene. This conjugate enone is the electro- cyclization yields the substrate for IkaC and another hydride-
phile for water addition to yield the 3’-ketoadenosine. Then initiated cyclization to yield the fused 5-6-5 core of the
back transfer of the hydride from bound NADH gives the bacterial metabolite ikarugamycin.[26b]
observed product adenosine. In addition to these NAD/NADH-dependent redox
Initiation of nucleophilic cascades can occur through an enzymes that set off cascade reactions, there are several
initial transfer of a hydride from NAD(P)H to an olefin in flavin-dependent enzymes that conduct redox catalysis that
a co-substrate, thereby creating a transient carbon nucleo- uncover cryptic reactivity in nascent products. Three exam-
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Scheme 5. Oxa-1,4-cojugate additions create pyran rings on the PKS assembly lines during pederin (A), ambruticin S (B), and salinomycin (C)
polyketide assembly.
ples are presented. The first is the FAD-enzyme Sol5 that shown. Collapse of the tetrahedral adduct generates an acid
converts the exocyclic alcohol group in prosolanapyrone II attached to the C-ring and an initial enediolate from what had
into the pyrone aldehyde in prosolanapyrone III in a fungus been the D-ring. Ketonization yields the hydroxyketone
that causes potato blight (Scheme 9).[27] The aldehyde is now grouping in the product mithramycin, with net oxygenative/
in conjugation with the adjacent olefin and that apparently hydrolytic fragmentation of the d-ring cyclohexenone.[28]
lowers the energy barrier so that it can act as a dienophile The third FAD enzyme, EncM, catalyzes oxidation of C4
towards the terminal diene. The result is an apparent Diels– in the side chain of the advanced polyketide chain tethered to
Alder [4+2] cyclization to the decalin bicycle in solanapyrone. the type II PKS EncC protein in enterocin biosynthesis.[29] A
The second enzyme is an FAD-containing monooxyge- notable aspect of the EncM catalytic mechanism is the
nase that converts the tetracyclic core of glycosylated discovery that it uses a newly identified flavin-N5-oxide as its
premithramycin B in to the tricyclic ring in mithramycin DK oxygen atom transfer catalyst to the 1,3-diketone methyl-
(Scheme 10). The enzyme MtmOIV is a Baeyer–Villiger ene[30] (Scheme 11). This creates a transient 1,2,3-triketo
catalyst, which delivers a nucleophilic oxygen to the d-ring moiety and sets in motion a Favorskii-type rearrangement,
enone with ring expansion to the lactone. The lactone as presumably generating a hydroxycyclopropanone intermedi-
a cyclic ester is labile to water-catalyzed ring opening as ate. That highly strained electrophile can be captured by an
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 6. The antibiotic lugdunin arises from an NRPS assembly line cascade. The release step involves reduction of tethered peptidyl thioester
by hydride transfer from NADH catalyzed by the LugC terminal reductase domain. The released aldehyde can circularize as the cyclic imine, which
is further driven to accumulate as the cyclic thiazolidine from addition of the cysteine thiolate to the imine.
Scheme 7. The enzyme SAH lyase starts a cascade leading to thioether cleavage by initial hydride-mediated oxidation of the ribose-3-OH to the
ketone. This lowers energy for C4 carbanion as the internal nucleophile required for C5 S cleavage.
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Two categories of electrophilic cascades are discussed in 4.2. Enzymatic Methylation as Initiator of a Cyclization Cascade
this section. The most extensive occurs in terpene biosyn-
thetic pathways in which carbocation chemistry dominates A variant of controlled induction of an electrophilic
enzymatic reaction sequences. In the second category, we cyclization occurs in the late stages of assembly of the
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 9. The redox step catalyzed by the FAD-containing solanapyrone synthase brings the aldehyde into conjugation with the exocyclic olefin
to increase its reactivity as a dienophile for the ensuing Diels–Alder cyclization.
Scheme 10. The FAD-enzyme that catalyzes a Baeyer–Villiger oxygenation on the d-ring of tetracyclic premithramycin converts the cylohexenone
D-ring into the ring-expanded lactone. This is now labile to water-mediated hydrolytic opening. The released enediolate isomerizes to the tricyclic
ketone mithramycin product.
prenylated indole teleocidin B, an inhibitor of protein kin- pling of the valyl nitrogen and C4 on the indole moiety.[41] To
ase C (Scheme 15).[39] The precursor of teleocidin B is the go from lyngbyatoxin A to the teleocidin B stereoisomers
indolactam lyngbyatoxin A, itself the causative agent of requires a new C C bond at C6 of the indole ring. Studies by
seaweed dermatitis.[40] The biosynthesis of lyngbyatoxin A is Abe and colleagues[39] revealed that the methyltransferase
understood to involve a two-module NRPS and then a P450 TleD is the enzyme that sets the appropriate chemical cascade
enzyme (thwarted oxygenase; see the section below on in motion. S-adenosylmethionine donates a [CH3+] equivalent
radical cascades), with a mechanism involving radical cou- to the terminal olefin at C25, creating the initial C26 tertiary
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Scheme 11. The EncM flavoenzyme doubly oxygenates a 1,3-diketo moiety of the enzyme-bound poly-b-keto intermediate to a 1,2,3-triketo nascent
product via the newly discovered flavin-N5-oxide cofactor. This product intermediate is subject to a Favorskii-type nucleophilic rearrangement
cascade with formation of a transient hydroxycyclopropanone.
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 13. A diterpene rearrangement cascade converts acyclic C20 geranylgeranyl-diphosphate into tricyclic taxadiene.
come from similar chemical logic in the active site of the epoxide by a vitamin B2-dependent squalene epoxidase. This
Mcl24 chloroperoxidase, in this case via route c) through epoxide is then the electrophilic substrate for oxidosqualene
a chloronium-ion assisted a-hydroxyketone rearrangement cyclase, with participation of the p electrons from four of the
(Scheme 16).[43] Thus Mcl24 single-handedly provides the olefins (not five as in squalene-hopene cyclase, presumably
three merochlorin natural product skeletons from the same because of folded conformer differences in the active sites) to
pre-merochlorin precursor. the protosterol cation (Scheme 14 B, see Ref. [13] for a sum-
Total syntheses of both merochlorins A and B have been mary).[47] A cascade of two hydride and two methyl migrations
achieved.[42b, 44] The four-enzyme pathway to merochlorins in occurs before cation quenching and release of tetracyclic
a single test tube has also been achieved,[45] as well as lanosterol. It has been argued that the squalene substrate in
a unifying hypothesis for naphthoquinone-containing mero- the hopene cyclase is an all pre-chair foldamer but is folded as
terpenoid scaffolds.[43, 46] Comparison of the strategic use of a pre-chair-boat-chair conformer in oxidosqualene cyclase.[47]
“Cl+” in this case versus “CH3+” in the teleocidin case above Correspondingly, a chair-chair-chair foldamer for squalene
shows the different tools available in enzymatic sites to epoxide would give the 6,6,6,6,6-pentacyclic scaffold of b-
initiate electrophilic cascade chemistry. amyrin, the most common steroid scaffold in plant metabo-
We will describe a different halogenation strategy in lism. Conversion of the initial dammarenyl cation via
Section 8, in which chlorine radical equivalents are employed a lupanyl cation into the oleanyl cation is followed by
in an enzymatic cascade to cyclindrocyclophane, and compare quenching through proton abstraction by a specifically placed
it to nonenzymatic metathesis synthetic strategy. enzymatic base (Scheme 14 C).[48]
While many of the steroid cyclases terminate the cation-
driven cascade rearrangements at one specific stage and
4.4. Tandem Epoxide Formation and Opening in Enzymatic region by proton abstraction or water addition, there are
Electrophilic Cascades family members that show some promiscuity. The baruol
synthase from the Arabidopsis plant generates baruol at
The more common modes of squalene enzymatic cycliza- nearly 90 % of the flux, with the remaining material distrib-
tion involve prior epoxidation of the 2,3-double bond to the b- uted over 22 minor products, varying in amounts from 0.02–
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Scheme 14. Triterpene cyclization cascades. A) Squalene-hopene cyclase initiates the electrocyclic cascade by protonation of the 2,3-terminal
double bond of the acyclic C30 hexaene squalene. B) Cyclization of squalene to lanosterol via a cascade of cation, hydride, and methyl group
migrations after oxidosqualene formation. C) Enzymatic cyclization cascade from 2,3-epoxysqualene to the pentacyclic b-amyrin product is the
predominant mode in plant metabolism.
2.7 % (Scheme 17).[49] This product range proves that a cas- indoline intermediate then undergoes capture at C2 of the
cade of multiple rearranged cationic species exist and can be indole ring by the neighboring olefin, loss of water, and
quenched, perhaps as 22 minor mistakes of timing and oxidative aromatization.[51d]
placement. The actions of PaxM and PaxB illustrate a widely used
Several examples of disappearing-epoxide strategies cascade strategy of tandem enzymes that make and then open
occur at the interface of indole alkaloid and prenyl transfer epoxides as “disappearing electrophiles” in cascade reac-
enzymology. One such example occurs during assembly of the tions.[52] The squalene epoxidase and oxidocyclase pair above
paxilline scaffold. Indole capture of the C20 GGPP yields 3- are such examples. So are Lsd18 and Lsd19 in lasalocid
geranylgeranyl indole (Scheme 18 A). This is the substrate for biosynthesis, but in that case the nucleophiles initiating the
PaxM, which bis-epoxidizes the terminal two of four side- cascades are not p electrons of olefins but instead the
chain olefins. The bis-epoxide is now subject to enzyme- alkoxide forms of internal -OH groups.[53] As shown in
mediated capture by p electrons of the remaining two double Scheme 19, Lsd19 acts on the bis-epoxide generated by Lsd18
bonds by the PaxB enzyme to create the hexacyclic frame- with regio- and stereospecific control to generate two sizes of
work of paspaline,[50] which upon subsequent double oxygen- cyclic ethers: the five-member dihydrofuran and the six-
ation serves as a precursor to the end product paxillin. A member dihydropyran. Although the enzymes remain to be
related strategy is used in assembly of indolosesquiterpenes, discovered for the dinoflagellate polyether toxins, such as the
represented by the pentacyclic family of xiamycin.[51] The 11-fused ether scaffold of brevetoxin B and 13-cyclic ethers in
conversion of farnesylindole into xiamyicin involves three ciguatoxins, it is anticipated that polyepoxide precursors will
kinds of enzymatic oxygenations (Scheme 18 B). The first is be converted in comparable cascades (Scheme 20).[54] Anal-
a prototypic epoxidation of the terminal olefin of farnesyl ogously, Leadlay and colleagues have presented evidence for
indole and electrophilic closure of two carbacyclic rings to MonC involvement as an epoxidase to create a triepoxide
form preindosespene. The second round of three P450- that, on opening by MonB, yields the five cyclic ethers in the
mediated oxygenations converts an exocyclic methyl into polyether antibiotic monensin.[55] The sequence of polyolefin
the carboxylate via an intermediate alcohol and aldehyde, to polyepoxide to fused pyran and furan ring scaffolds in
consuming three molecules of O2. C C bond formation to cyclic polyethers was predicted by Cane, Celmer, and West-
yield the xiamycin scaffold occurs by flavoenzyme action ley,[53b] and further refined by Spencer and colleagues[55b] for
through a cryptic indole ring hydroxylation. The resultant monensin.
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 15. Enzymatic C-methylation at an olefin initiates an electrophilic cascade in teleocidin B biosynthesis.
Scheme 16. Enzymatic double chlorination of a naphthol ring by chloronium ion equivalents sets a carbocation cascade in motion in
merochlorin A and B formation. Merochlorin C formation rather involves a Cl+-mediated a-hydroxyketone rearrangement and a third chlorination
event.
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Scheme 17. Baruol synthase leaks a set of minor products reflecting capture of intermediates at different points in the cationic cascade process.
Bold arrows show the primary route to baruol with representative pathway byproducts shown to other cyclic triterpenes with relative percentage
product distribution.
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 18. Disappearing-epoxide cascade reactions in the enzymatic conversion of 3-prenylindole substrates into paspaline (A) and xiamycin (B).
asynchronous).[57] Certainly, the 4+2 conversions dramatically SpnG, SpnL then cyclizes the remaining diene to the cyclo-
alter framework connectivity in the enzymatic products.[58] pentene ring in what may be a Rauhut–Currier-type of C C
Some of them, we would put in the cascade category. bond formation.[60] The two enzymes SpnF and SpnL thus
In connection with this, we highlight the tandem action of build the constrained 5,6,5-tricyclic core of spinosyn A
the enzymes PyrE3 and PyrE4 in the pathway to pyrroindo- (Scheme 23). Energetic calculations for the SpnF transition
mycin B (Scheme 22).[59] It is not so much that either enzyme states support a [4+2] reaction but also suggest a possible
by itself is a cascade catalyst but rather that tandem action [6+4] route that would need to be followed by a 3,3-Cope
leads to the generation of the pentacyclic core of the natural rearrangement to give the observed product.[60]
product. Furthermore, the two enzymes illustrate the two The Diels–Alder [4+2] cyclization was originally de-
variants of the kinds of 4+2 outcomes that have been scribed in 1928.[61] An oxo-Diels–Alder reaction to give
catalogued to date in enzyme systems. PyrE3 is a “typical” dihydropyran derivatives was reported in 1949.[62] The corre-
decalin-forming [4+2] catalyst. The next enzyme, PyrE4, sponding intramolecular aza-Diels–Alder reaction on an
takes this dialkyl decalin product and makes a spirotetronate imino alkyne was reported in 2009.[63] The biosyntheses of
ring system as the pentacyclic scaffold is constructed through around 80 members of a peptide antibiotic class that has been
the second type of apparent [4+2] cyclization. morphed into products with a central 2,4,6-trithiazolylpyr-
With some degree of analogy, the insecticide molecule idine embedded in a peptide macrocycle have recently been
spinosyn A has a 5,6,5-tricyclic core that is also of pericyclic formulated to involve an aza-Diels–Alder-type reaction
origin. SpnF has been purified to homogeneity and shown to catalyzed by the purified enzyme ThiM (Scheme 24).[64] The
accelerate a slow non-enzymatic cyclization of the macro- total synthesis of thiocillin and several analogues was
cyclic substrate to the central cyclohexene and the right-hand reported more than a decade ago, and aza-Diels–Alder and
cyclopentane rings. Following enzymatic glycosylation by aza-Mannich chemistry was at the heart of the approaches,
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 23. SpnF and SpnL build the fused 5,6,5-tricyclic core of the
Scheme 25. The enzyme LepI carries out both a conventional [4+2]
insecticidal agent spinosyn through Diels–Alder and Rauhut–Currier
cyclization and an oxa-[4+2] cyclization competitively. The latter
reactions.
reaction yields the main product leporin C. The spiro product from the
conventional [4+2] pathway is then subjected to an enzymatic [3,3]
An example of an oxo-Diels–Alder enzymatic reaction retro-Claisen reaction to rescue the stranded material and convert it
with an additional novel twist has recently been reported for into leporin C.
Scheme 24. Biosynthesis of the trithiazolylpyridine core of thiocillin-type antibiotics. Proposed aza-Diels–Alder cyclization in the reaction catalyzed
by ThiM during thiocillin assembly. Creation of the pyridine ring at the core of the trithiazolylpyridine array also closes the 26-membered
macrocyclic ring in thiocillin.
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Alder reaction would produce the observed leporin C product by female gametes of marine brown algae. Among the
directly. On the other hand, a conventional [4+2]-catalyzed plethora of related algal cycloalkenes is ectocarpene.[70] This
pathway would yield the indicated spirodecalin. Indeed, this C11 cycloheptadiene is thought to arise from oxygenative
was observed as a co-product with leporin C at early time fragmentation of the D5,8,11,14,17 penta-unsaturated C20 fatty
points of substrate conversion. Later, the enzyme utilized this acid primary metabolite by radical-based peroxide chemistry
co-product and converted it all into leporin C. This is most to the indicated two fragments. Isotope labeling studies are
readily formulated as a retro-Claisen rearrangement consistent with this proposed mechanism but there are no
(Scheme 25). This direction for a 3,3 conversion is the first reports on the enzymatic details (Scheme 27). The 3,3-
seen in an enzyme-catalyzed reaction. In summary, LepI rearrangement step would occur from the indicated trienyl
appears to be able to conduct both a conventional [4+2] and cyclopropane pheromone, detected as an unstable intermedi-
a hetero-[4+2] cyclization reaction competitively (ambivalent ate, and that may be a nonenzymatic thermal reaction at room
transition state), and then retrieve the stranded nascent temperature. It certainly qualifies as a biosynthetic cascade
Diels–Alder product through a retro-Claisen reaction in reaction.
a remarkable cascade of pericyclic transformations. A novel
feature of the LepI enzyme is that it requires the cofactor
SAM, not for its methylating capacity, but rather to use the
positive charge of SAM to direct the pericyclic reactions.
The forward 3,3-Claisen rearrangement of ally vinyl
ethers has been known for decades to be the mechanism of
the key enzymatic transformation in aromatic amino acid
biosynthesis in microbes and plants (Scheme 26). Chorismate
Scheme 26. Chorismate mutase catalyzes the only known [3,3]-rear- Scheme 27. Proposed Cope rearrangement in the biosynthesis of the
rangement in primary metabolism of microbes and plants. brown algal feeding deterrent ectocarpene from a transient cyclo-
propane pheromone, which in turn arises from peroxidative fragmenta-
tion of a polyunsaturated fatty acyl peroxide.
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
6. Radical-Based Cascades
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
oxidation state while the nonheme enzymes reach an FeIV=O downstream partner desacetoxycephalosporin-C synthase
oxidation state. Both high-valent oxo-iron species can cleave (DAOCS) in the formation of these major antibiotic classes
unactivated C H bonds in bound substrates homolytically to (Scheme 30).[78, 80] These O2-reducing nonheme iron oxygen-
yield the substrate radical and Fe(IV/III) OH. For a prototypic ases dramatically alter their substrates in cascade reactions.
monoxygenase outcome, transfer of OHC to substrateC yields IPNS converts the acyclic tripeptide aminoadipoyl-l-cys-
hydroxylated product and regenerates either the FeIII-heme teinyl-d-valine into the fused 4,5-warhead of isopenicillin.
or FeII nonheme resting state for the next catalytic cycle. The l-amino group of the aminoadipoyl side chain is next
There have been many dozens of examples of such hydrox- epimerized by an enzyme to penicillin N. Deep dissections of
ylases reported.[78] (See Ref. [13] for examples in natural the mechanism of both IPNS and DAOCS mechanisms have
product biosynthesis). Some of these oxygenases act itera- validated the radical-based cascades.[81]
tively, for example, on substrate methyl groups to carry out For IPNS, the b-lactam ring is formed first and then the
three steps of two-electron oxidations each, to go from fused five-member thiane ring is constructed, without loss of
a methyl group to an alcohol to an aldehyde to a carboxylic the monocyclic lactam intermediate. Carbon-based and
acid. By some criteria, these could be considered cascade sulfur-based odd-electron intermediates have been proposed.
reactions, but they are not the subject of this discussion. DAOCS in turn uses a high-valent FeIV=O intermediate to
Rather, a significant subset of catalysts in these enzyme cleave a C H bond of penicillin N homolytically at one of the
superfamilies never complete the OHC rebound step.[13, 79] An two prochiral CH3 substituents in the 5-member thiane ring.
alternate fate of the carbon-centered radical intervenes, One can formulate the set of rearrangements as proceeding
typically in a rearrangement cascade. The released product through the 3-member episulfide radical that opens to give
has not incorporated an oxygen atom. Both atoms from the the ring expansion, followed by transfer of HC back to FeIII to
co-substrate O2 end up as in H2O molecules. These enzymes, regenerate starting FeII. The final expanded product has a 6-
we term “thwarted oxygenases”: O2 is reductively activated membered sulfur-containing ring with a C=C double bond
and fragmented, high-valent iron-oxo species are formed as fused to the beta lactam—the hallmark of the cephalosporin
strong oxidants, substrate C H bonds are cleaved homolyti- warheads.
cally, but the oxygen transfer step (OHC rebound) is not Many examples of “thwarted oxygenase outcomes” are
completed. found that reflect cascade intramolecular reactions of bound
The most famous of these rerouted oxygenases are substrate radicals that effectively outcompete the intermo-
perhaps isopenicillin N synthase (IPNS) and its immediate lecular transfer of OHC. Among them are transformation of
Scheme 30. Isopenicillin N synthase (IPNS) is a “thwarted oxygenase” that converts the acyclic tripeptide ACV into the fused 4,5-ring system of
the penicillin family of b-lactam antibiotics through a cascade of radicals.
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Scheme 32. The P450-containing GsfF enzyme catalyzes a radical-based reaction pathway to create the spirocyclic ring system of the fungal
metabolite griseofulvin in a “thwarted oxygenase” mode.
Scheme 33. Additional enzyme-mediated radical redirection reactions in the biosynthesis of the alkaloid salutaridine (A), the indolecarbazoles
staurosporine and rebeccamycin (B), and the diketopiperazine fumitremorgin C (C).
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Scheme 35. P450 CYP88A-mediated conversion of ent-kaurenoic acid into giberellic acid-12-aldehyde via a reaction sequence onvloving a radical-
based ring contraction and aldehyde carbon -C6HC(OH) extrusion concomitant with oxygen transfer.
Scheme 36. Enzyme mediated O2-dependent radical cascades in the late stages of the biosynthesis of anditomin meroterpenoids. AndA creates
a bicyclooctane ring embedded in the meroterpenoid scaffold, while AndF introduces the final nonoxygenative radical rearrangements in the 12-
enzyme pathway to anditomin.
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
6.1.2. Okaramine: One Satisfied and Two Thwarted Oxygenases N H bond homolytically (Scheme 37). Rather than complet-
in a Pathway That Builds an Octacyclic Scaffold in Five ing a net N-hydroxylation event, the nitrogen radical reacts
Steps with the nearby terminal olefin to form a new C N bond. This
olefin-radical coupling is analogous to the step noted for
Penicillium molds are famous for their biosynthetic AndA above. In this case, it creates an eight-membered
capacity to build heterocyclic scaffolds, among them of heterocycle, an azocine ring radical that can be reduced by
course the penicillins and cephalosporins. Recently the path- one electron/HC transfer to the azocine.
way to the octacyclic framework of the fungal metabolite Now the third “oxygenase”, OkaE, goes to work
okaramine E, an insect ion-channel blocker, has been shown (Scheme 37). It acts on the other end of the substrate,
to occur in a short efficient five-enzyme pathway from the cleaving the C H bond between the two five-member
primary metabolites l-tryptophan and D2-isopentenyl diphos- pyrroline rings homolytically to generate the indicated
phate (Scheme 37).[93] The first enzyme is a two-module carbon radical. As with the previous enzyme, this carbon-
NRPS assembly line, which condenses two molecules of l- centered radical can react with the neighboring prenyl-group-
tryptophan and mediates intramolecular capture by the amine derived olefin, in this case to form a four-member azetidine
of the second tryptophan residue to release the cyclic ring radical. One electron/HC reduction completes the five-
diketopiperazine. A prenyltransferase then asymmetrically step formation of the octacyclic scaffold of okaramine E, with
prenylates one indole moiety at N1 and the other at C2 with new 4-ring, 5-ring and 8-ring fused heterocycles from three
capture of the allyl cations at their C3 rather than C1 centers to cascades. Oxygen delivery to substrate is kinetically compe-
give “reversed” regiochemistry of prenylation, which is tent in only the first of the three oxygenases and emphasizes
important for subsequent cascades with the last two enzymes the different roles for flavoenzymes versus iron enzyme
in the pathway to okaramine E. These two steps set up the oxygenases.
action of three oxygenases that represent the three known
monoxoygenase types: a flavoenzyme, a P450 heme iron- 6.1.3. Phenylpropanoid Thwarted Oxygenases
containing enzyme, and a mononuclear nonheme iron
enzyme. A final two examples, of many that could be cited (see
The latter two are the “thwarted oxygenases” while the Ref. [13], Chapter 7), come from plant phenylpropanoid
FAD enzyme, OkaB, actually acts to epoxidize the indole natural product pathways. One is the dimerization of coniferyl
moiety that is prenylated at N1. This reaction sets off a short alcohol to a set of three distinct regioisomeric dimers, with
cascade in which the adjacent amide NH of the diketopiper- dramatically different 8-8’ connectivities (Scheme 38 A).[94]
azine, weak nucleophile that it is, opens the epoxide regio- Pinoresinol is the precursor to a variety of downstream
and stereospecifically to create a new cyclopentane ring natural products in plants,[95] including sesamin and podo-
interposed between the indole and the diketopiperazine in phyllotoxin. A second classic case is the enzymatic conversion
a fused tetracyclic array in okaramine C. of flavonoid to isoflavonoid scaffolds in plant secondary
This is the substrate for OkaD, the P450 enzyme. It uses metabolism, as exemplified by the O2- and P450-mediated
the FeV=O oxidant to cleave the remaining diketopiperazine conversion of naringenin into genistein,[96] itself a precursor to
Scheme 37. Enzymatic assembly of the octacyclic framework of the insect ion channel blocker okaramine E in a short, efficient pathway involving
two thwarted oxygenases, the P450 OkaD and the nonheme iron enzyme OkaE, that catalyze radical cascade reactions to generate the eight-
member azocine ring and the four-member azetidine ring, respectively.
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Scheme 39. Two radical SAM enzymes in the anaerobic aminofutalosine pathway to menaquinone (vitamin K).
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Scheme 41. Synthesis and biosynthesis of the 7,7-para-cyclophane cylindrocyclophane F by different cascade strategies. A) Synthesis by a double
metathesis cascade. B) Biosynthesis by an apparent Friedel Crafts type bis alkylation cascade to create the para-cyclophane macrocycle.
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
[21] C. T. Walsh, S. W. Haynes, B. D. Ames, X. Gao, Y. Tang, ACS [42] a) L. Kaysser, P. Bernhardt, S. J. Nam, S. Loesgen, J. G. Ruby, P.
Chem. Biol. 2013, 8, 1366 – 1382. Skewes-Cox, P. R. Jensen, W. Fenical, B. S. Moore, J. Am.
[22] a) P. Pçplau, S. Frank, B. I. Morinaka, J. Piel, Angew. Chem. Int. Chem. Soc. 2012, 134, 11988 – 11991; b) S. Diethelm, R. Teufel,
Ed. 2013, 52, 13215 – 13218; Angew. Chem. 2013, 125, 13457 – L. Kaysser, B. S. Moore, Angew. Chem. Int. Ed. 2014, 53,
13460; b) G. Berkhan, F. Hahn, Angew. Chem. Int. Ed. 2014, 53, 11023 – 11026; Angew. Chem. 2014, 126, 11203 – 11206.
14240 – 14244; Angew. Chem. 2014, 126, 14464 – 14468; c) H. [43] Z. D. Miles, S. Diethelm, H. P. Pepper, D. M. Huang, J. H.
Luhavaya, M. V. Dias, S. R. Williams, H. Hong, L. G. de Oli- George, B. S. Moore, Nat. Chem. 2017, 9, 1235 – 1242.
veira, P. F. Leadlay, Angew. Chem. Int. Ed. 2015, 54, 13622 – [44] a) H. P. Pepper, J. H. George, Angew. Chem. Int. Ed. 2013, 52,
13625; Angew. Chem. 2015, 127, 13826 – 13829. 12170 – 12173; Angew. Chem. 2013, 125, 12392 – 12395; b) R.
[23] A. Zipperer, M. C. Konnerth, C. Laux, A. Berscheid, D. Janek, Meier, S. Strych, D. Trauner, J. Org. Chem. 2014, 16, 2634 –
M. Marschal, M. Burian, N. A. Schilling, C. Slavetinsky, C. 2637.
Weidenmaier, M. Willman, H. Kahlbacher, B. Schittek, H. [45] R. Teufel, L. Kaysser, M. T. Villaume, S. Diethelm, M. K.
Brotz-Osterhelt, S. Grond, A. Peschel, B. Krismer, Nature 2016, Carbullido, P. S. Baran, B. S. Moore, Angew. Chem. Int. Ed.
535, 511 – 516. 2014, 53, 11019 – 11022; Angew. Chem. 2014, 126, 11199 – 11202.
[24] J. L. Palmer, R. H. Abeles, J. Biol. Chem. 1979, 254, 1217 – 1226. [46] B. S. Moore, Synlett 2017, 29, 401 – 409.
[25] a) F. Geu-Flores, N. Sherden, V. Courdavault, V. Burlat, W. [47] K. U. Wendt, Angew. Chem. Int. Ed. 2005, 44, 3966 – 3971;
Glenn, C. Wu, E. Nims, Y. Cui, S. OConnor, Nature 2012, 492, Angew. Chem. 2005, 117, 4032 – 4037.
138 – 142; b) H. Kries, L. Caputi, C. Stevenson, M. Kamileen, N. [48] a) R. Thimmappa, K. Geisler, T. Louveau, P. OMaille, A.
Sherden, F. Geu-Flores, D. Lawson, S. OConnor, Nat. Chem. Osbourn, Annu. Rev. Plant Biol. 2014, 65, 225 – 257; b) T.
Biol. 2016, 12, 6 – 8. Hoshino, Org. Biomol. Chem. 2017, 15, 2869 – 2891.
[26] a) J. Antosch, F. Schaefers, T. Gulder, Angew. Chem. Int. Ed. [49] S. Lodeiro, Q. Xiong, W. K. Wilson, M. D. Kolesnikova, C. S.
2014, 53, 3011 – 3014; Angew. Chem. 2014, 126, 3055 – 3058; Onak, S. P. Matsuda, J. Am. Chem. Soc. 2007, 129, 11213 –
b) G. Zhang, W. Zhang, Q. Zhang, T. Shi, L. Ma, Y. Zhu, S. Li, 11222.
H. Zhang, Y. Zhao, R. Shi, C. Zhang, Angew. Chem. Int. Ed. [50] K. Tagami, C. Liu, A. Minami, M. Noike, T. Isaka, S. Fueki, Y.
2014, 53, 4840 – 4844; Angew. Chem. 2014, 126, 4940 – 4944. Shichijo, H. Toshima, K. Gomi, T. Dairi, H. Oikawa, J. Am.
[27] a) K. Kasahara, T. Miyamoto, T. Fujimoto, H. Oguri, T. Chem. Soc. 2013, 135, 1260 – 1263.
Tokiwano, H. Oikawa, Y. Ebizuka, I. Fujii, ChemBioChem [51] a) H. Li, Q. Zhang, S. Li, Y. Zhu, G. Zhang, H. Zhang, X. Tian,
S. Zhang, J. Ju, C. Zhang, J. Am. Chem. Soc. 2012, 134, 8996 –
2010, 11, 1245 – 1252; b) W. Kim, C. M. Park, J. J. Park, H. O.
9005; b) Z. Xu, M. Baunach, L. Ding, C. Hertweck, Angew.
Akamatsu, T. L. Peever, M. Xian, D. R. Gang, G. Vandemark,
Chem. Int. Ed. 2012, 51, 10293 – 10297; Angew. Chem. 2012,
W. Chen, Mol. Plant-Microbe Interact. 2015, 28, 482 – 496.
124, 10439 – 10443; c) H. Li, Y. Sun, Q. Zhang, Y. Zhu, S. M. Li,
[28] M. P. Beam, M. A. Bosserman, N. Noinaj, M. Whenkel, J. Rohr,
C. Zhang, Org. Lett. 2015, 17, 306 – 309; d) S. Kugel, M.
Biochemistry 2009, 48, 4476 – 4487.
Baunach, P. Baer, M. Ishida-Ito, S. Sundaram, Z. Xu, M. Groll,
[29] a) L. Xiang, J. A. Kalaitzis, B. S. Moore, Proc. Natl. Acad. Sci.
C. Hertweck, Nat. Commun. 2017, 8, 15804.
USA 2004, 101, 15609 – 15614; b) R. Teufel, A. Miyanaga, Q.
[52] a) M. C. Tang, Y. Zhou, K. Watanabe, C. T. Walsh, Y. Tang,
Michaudel, F. Stull, G. Louie, J. P. Noel, P. S. Baran, B. Palfey,
Chem. Rev. 2017, 117, 5226 – 5333; b) C. T. Walsh, Y. Tang,
B. S. Moore, Nature 2013, 503, 552 – 556.
Biochemistry 2018, 57, 3087 – 3104.
[30] a) R. Teufel, F. Stull, M. J. Meehan, Q. Michaudel, P. C.
[53] a) A. Minami, M. Shimaya, G. Suzuki, A. Migita, S. Shinde, K.
Dorrestein, B. Palfrey, B. S. Moore, J. Am. Chem. Soc. 2015,
Sato, K. Watanabe, T. Tamura, H. Ogura, H. Oikawa, J. Am.
137, 8078 – 8085; b) R. Saleem-Batcha, F. Stull, J. N. Sanders, Chem. Soc. 2012, 134, 7246 – 7249; b) D. E. Cane, W. D. Celmer,
B. S. Moore, B. A. Palfey, K. N. Houk, R. Teufel, Proc. Natl. J. W. Westley, J. Am. Chem. Soc. 1983, 105, 3594 – 3600; c) T.
Acad. Sci. USA 2018, 115, 4909 – 4914. Liu, D. E. Cane, Z. Deng, Methods Enzymol. 2009, 459, 187 –
[31] a) Q. Cheng, L. Xiang, M. Izumikawa, D. Meluzzi, B. S. Moore, 214; d) A. R. Gallimore, Nat. Prod. Rep. 2009, 26, 266 – 280.
Nat. Chem. Biol. 2007, 3, 557 – 558; b) J. A. Kalaitzis, Q. Cheng, [54] R. Kellmann, A. Stken, R. J. Orr, H. M. Svendsen, K. S.
P. M. Thomas, N. L. Kelleher, B. S. Moore, J. Nat. Prod. 2009, Jakobsen, Mar. Drugs 2010, 8, 1011 – 1048.
72, 469 – 472. [55] a) M. Oliynyk, C. B. Stark, A. Bhatt, M. A. Jones, Z. A.
[32] B. T. Ueberbacher, M. Hall, K. Faber, Nat. Prod. Rep. 2012, 29, Hughes-Thomas, C. Wilkinson, Z. Oliynyk, Y. Demydchuk, J.
337 – 350. Staunton, P. F. Leadlay, Mol. Microbiol. 2003, 49, 1179 – 1190;
[33] D. W. Christianson, Chem. Rev. 2017, 117, 11570 – 11648. b) A. R. Gallimore, C. B. W. Stark, A. Bhatt, B. M. Harvey, Y.
[34] C. A. Lesburg, G. Zhai, D. E. Cane, D. W. Christianson, Science Demydchuk, V. Bolanos-Garcia, D. J. Fowler, J. F. Staunton,
1997, 277, 1820 – 1824. P. F. Leadlay, J. B. Spencer, Chem. Biol. 2006, 13, 453 – 460.
[35] C. N. Tetzlaff, Z. You, D. E. Cane, S. Takamtsu, S. Omura, H. [56] X. M. Mao, Z. J. Zhan, M. N. Grayson, M. C. Tang, W. Xu,
Ikeda, Biochemistry 2006, 45, 6179 – 6186. Y. Q. Li, W. B. Yin, H. C. Lin, Y. H. Chooi, K. N. Houk, Y.
[36] K. M. Y. Jin, R. M. Coates, R. Croteau, D. W. Christianson, Tang, J. Am. Chem. Soc. 2015, 137, 11904 – 11907.
Nature 2011, 469, 116 – 120. [57] B. S. Jeon, S. A. Wang, M. Ruszczcky, H. W. Liu, Chem. Rev.
[37] J. Guerra-Bubb, R. Croteau, R. M. Williams, Nat. Prod. Rep. 2017, 117, 5367 – 5388.
2012, 29, 683 – 696. [58] A. Minami, H. Oikawa, J. Antibiot. 2016, 69, 500 – 506.
[38] a) G. Siedenburg, D. Jendrossek, Appl. Environ. Microbiol. [59] Z. Tian, P. Sun, Y. Zan, Z. Wu, Q. Zheng, S. Zhou, H. Zhang, F.
2011, 77, 3905 – 3915; b) K. U. Wendt, K. Poralla, G. E. Schulz, Yu, X. Jia, D. Chen, A. Mandi, T. Kurtan, W. Liu, Nat. Chem.
Science 1997, 277, 1811 – 1815. Biol. 2015, 11, 259 – 265.
[39] T. Awakawa, L. Zhang, T. Wakimoto, S. Hoshino, T. Mori, T. [60] B. S. Jeon, M. W. Ruszczcky, W. K. Russell, G. M. Lin, N. Kim,
Ito, J. Ishikawa, M. Tanner, I. Abe, J. Am. Chem. Soc. 2014, 136, S. H. Choi, S. A. Wang, Y. N. Liu, D. H. Russell, K. N. Houk,
9910 – 9913. H. W. Liu, Proc. Natl. Acad. Sci. USA 2017, 114, 10408 – 10413.
[40] J. H. Cardellina, F. J. Marner, R. E. Moore, Science 1979, 204, [61] O. Diels, K. Alder, Liebigs Ann. Chem. 1928, 460, 98 – 122.
193 – 195. [62] T. L. Gresham, T. R. Steadman, J. Am. Chem. Soc. 1949, 71,
[41] S. E. Ongley, X. Bian, Y. Zhang, R. Chau, W. H. Gerwick, R. 737 – 738.
Muller, B. A. Neilan, ACS Chem. Biol. 2013, 8, 1888 – 1893. [63] S. Desrat, P. van de Wege, J. Org. Chem. 2009, 74, 6728 – 6734.
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
[64] W. J. Wever, J. W. Bogart, J. A. Baccile, A. N. Chan, F. C. [84] a) A. Gesell, M. Rolf, J. Ziegler, M. L. Daz Chvez, F. C.
Schroeder, A. A. Bowers, J. Am. Chem. Soc. 2015, 137, 3494 – Huang, T. M. Kutchan, J. Biol. Chem. 2009, 284, 24432 – 24442;
3497. b) S. Galanie, K. Thodey, I. J. Trenchard, M. Filsinger Inter-
[65] C. T. Walsh, M. G. Acker, A. A. Bowers, J. Biol. Chem. 2010, rante, C. D. Smolke, Science 2015, 349, 1095 – 1100.
285, 27525 – 27531. [85] a) A. R. Howard-Jones, C. T. Walsh, J. Am. Chem. Soc. 2007,
[66] D. P. Cogan, G. A. Hudson, Z. Zhang, T. V. Pogorelov, W. A. 129, 11016 – 11017; b) M. Makino, H. Sugimoto, Y. Shiro, S.
van der Donk, D. A. Mitchell, S. K. Nair, Proc. Natl. Acad. Sci. Asamizu, H. Onaka, S. Nagano, Proc. Natl. Acad. Sci. USA
USA 2017, 114, 12928 – 12933. 2007, 104, 11591 – 11596.
[67] M. Ohashi, F. Liu, Y. Hai, M. Chen, M. C. Tang, Z. Yang, M. [86] N. Kato, H. Suzuki, H. Takagi, Y. Asami, H. Kakeya, M.
Sato, K. Watanabe, K. N. Houk, Y. Tang, Nature 2017, 549, 502 – Uramoto, T. Usui, S. Takahashi, Y. Sugimoto, H. Osada,
506. ChemBioChem 2009, 10, 920 – 928.
[68] A. Y. Lee, J. D. Stewart, J. Clardy, B. Ganem, Chem. Biol. 1995, [87] H. C. Lin, T. McMahon, A. Patl, M. Corsello, A. Simon, W. Xu,
2, 195 – 203. M. Zhao, K. N. Houk, N. K. Garg, Y. Tang, J. Am. Chem. Soc.
[69] L. Y. P. Luk, Q. Qian, M. E. Tanner, J. Am. Chem. Soc. 2011, 2016, 138, 4002 – 4005.
133, 12342 – 12345. [88] H. C. Lin, G. Chiou, Y. H. Chooi, T. C. McMahon, W. Xu, N. K.
[70] W. Boland, Proc. Natl. Acad. Sci. USA 1995, 92, 37 – 43. Garg, Y. Tang, Angew. Chem. Int. Ed. 2015, 54, 3004 – 3007;
[71] a) S. Li, A. N. Lowell, F. Yu, A. Taveh, S. A. Newmister, N. Angew. Chem. 2015, 127, 3047 – 3050.
Baiir, J. M. Schaub, R. M. Williams, D. H. Sherman, J. Am. [89] S. P. Lathrop, M. Pompeo, W. T. Chang, M. Movassaghi, J. Am.
Chem. Soc. 2015, 137, 15366 – 15369; b) S. Li, A. N. Lowell, Chem. Soc. 2016, 138, 7763 – 7769.
S. A. Newmister, F. Yu, R. M. Williams, D. H. Sherman, Nat. [90] C. A. Helliwell, P. M. Chandler, A. Poole, E. S. Dennis, W. J.
Chem. Biol. 2017, 13, 467 – 469; c) Q. Zhu, X. Liu, Chem. Peacock, Proc. Natl. Acad. Sci. USA 2001, 98, 2065 – 2070.
Commun. 2017, 53, 2826 – 2829; d) Q. Zhu, X. Liu, Angew. [91] a) Y. Matsuda, T. Wakimoto, T. Mori, T. Awakawa, I. Abe, J.
Chem. Int. Ed. 2017, 56, 9062 – 9066; Angew. Chem. 2017, 129, Am. Chem. Soc. 2014, 136, 15326 – 15336; b) Y. Nakashima, T.
9190 – 9194. Mitsihashi, Y. Matsuda, M. Senda, H. Sato, M. Yamazaki, M.
[72] R. E. Moore, C. Cheuk, X. Qiang, G. M. L. Patterson, R. Uchiyama, T. Senda, I. Abe, J. Am. Chem. Soc. 2018, 140, 9743 –
9750.
Bonjouklian, T. A. Smitka, J. S. Mynderse, R. F. Foster, N. D.
[92] a) S. Salazar-Cerezo, N. Martnez-Montiel, J. Garca-Snchez,
Jones, J. K. Swartzendruber, J. B. Deeter, J. Org. Chem. 1987,
R. Prez-Y-Terrn, R. D. Martnez-Contreras, Microbiol. Res.
52, 1036 – 1043.
2018, 208, 85 – 98; b) B. Tudzynski, Appl. Microbiol. Biotechnol.
[73] S. A. Newmister, S. Li, M. Garcia-Borras, J. N. Sanders, S. Yang,
2005, 66, 597 – 611; c) R. S. Nett, M. Montanares, A. Marcassa,
A. N. Lowell, F. Yu, J. L. Smith, R. M. Williams, K. N. Houk,
X. Lu, R. Nagel, T. C. Charles, P. Hedden, M. C. Rojas, R. J.
D. H. Sherman, Nat. Chem. Biol. 2018, 14, 345 – 351.
Peters, Nat. Chem. Biol. 2017, 13, 69 – 74.
[74] a) C. Krebs, D. Galonić Fujimori, C. T. Walsh, J. M. J. Bollinger,
[93] C. Y. Lai, I. W. Lo, R. T. Hewage, Y. C. Chen, C. T. Chen, C. F.
Acc. Chem. Res. 2007, 40, 484 – 492; b) H. Nakamura, Y.
Lee, S. Lin, M. C. Tang, H. C. Lin, Angew. Chem. Int. Ed. 2017,
Matsuda, I. Abe, Nat. Prod. Rep. 2018, 35, 633 – 645.
56, 9478 – 9482; Angew. Chem. 2017, 129, 9606 – 9610.
[75] B. J. Landgraf, E. L. McCarthy, S. J. Booker, Annu. Rev.
[94] a) L. B. Davin, H. B. Wang, A. L. Crowell, D. L. Bedgar, D. M.
Biochem. 2016, 85, 485 – 514.
Martin, S. Sarkanen, N. G. Lewis, Science 1997, 275, 362 – 366;
[76] a) Q. Zhang, W. A. van der Donk, W. Liu, Acc. Chem. Res.
b) B. Pickel, A. Schaller, Appl. Microbiol. Biotechnol. 2013, 97,
2012, 45, 555 – 564; b) K. Yokoyama, E. A. Lilla, Nat. Prod. 8427 – 8438.
Rep. 2018, 35, 660 – 694. [95] R. B. Teponno, S. Kusari, M. Spiteller, Nat. Prod. Rep. 2016, 33,
[77] M. W. Ruszczycky, A. Zhong, H. W. Liu, Nat. Prod. Rep. 2018, 1044 – 1092.
35, 615 – 621. [96] W. Jung, O. Yu, S. M. Lau, D. P. OKeefe, J. Odell, G. Fader, B.
[78] C. J. Schofield, Z. Zhang, Curr. Opin. Struct. Biol. 1999, 9, 722 – McGonigle, Nat. Biotechnol. 2000, 18, 208 – 212.
731. [97] J. Broderick, B. R. Duffus, K. D. Duschene, E. M. Shepard,
[79] M. Mizutani, F. Sato, Arch. Biochem. Biophys. 2011, 507, 194 – Chem. Rev. 2014, 114, 4229 – 4317.
203. [98] B. W. Lepore, F. J. Ruzicka, P. A. Frey, D. Ringe, Proc. Natl.
[80] R. B. Hamed, J. R. Gomez-Castellanos, H. L. Ducho, M. A. Acad. Sci. USA 2005, 102, 13819 – 13824.
McDonough, C. J. Schofield, Nat. Prod. Rep. 2013, 30, 21 – 107. [99] C. J. Fugate, J. T. Jarrett, Biochim. Biophys. Acta Proteins
[81] a) P. L. Roach, I. J. Clifton, C. M. Hensgens, N. Shibata, C. J. Proteomics 2012, 1824, 1213 – 1222.
Schofield, J. Hajdu, J. E. Baldwin, Nature 1997, 387, 827 – 830; [100] M. I. McLaughlin, N. D. Lanz, P. J. Goldman, K. H. Lee, S. J.
b) K. Valegrd, A. C. van Scheltinga, M. D. Lloyd, T. Hara, S. Booker, C. L. Drennan, Proc. Natl. Acad. Sci. USA 2016, 113,
Ramaswamy, A. Perrakis, A. Thompson, H. J. Lee, J. E. 9446 – 9450.
Baldwin, C. J. Schofield, J. Hajdu, I. Andersson, Nature 1998, [101] T. Hiratsuka, K. Furihata, J. Ishikawa, H. Yamashita, N. Itoh, H.
394, 805 – 809. Seto, T. Dairi, Science 2008, 321, 1670 – 1673.
[82] a) O. Pylypenko, F. Vitali, K. Zerbe, J. A. Robinson, I. [102] a) N. Mahanta, D. Fedoseyenko, D. Dairi, T. P. Begley, J. Am.
Schlichting, J. Biol. Chem. 2003, 278, 46727 – 46733; b) K. Chem. Soc. 2013, 135, 15318 – 15321; b) S. Joshi, N. Mahanta, D.
Zerbe, K. Woithe, D. B. Li, F. Vitali, L. Bigler, J. A. Robinson, Fedoseyenko, H. Williams, T. P. Begley, J. Am. Chem. Soc. 2017,
Angew. Chem. Int. Ed. 2004, 43, 6709 – 6713; Angew. Chem. 139, 10952 – 10955.
2004, 116, 6877 – 6881; c) K. Woithe, N. Geib, K. Zerbe, D. B. [103] L. E. Cooper, D. Fedoseyenko, S. H. Abdelwahed, S. H. Kim, T.
Li, M. Heck, S. Fournier-Rousset, O. Meyer, F. Vitali, N. Dairi, T. P. Begley, Biochemistry 2013, 52, 4592 – 4594.
Matoba, K. Abou-Hadeed, J. A. Robinson, J. Am. Chem. Soc. [104] A. P. Mehta, S. H. Abdelwahed, N. Mahanta, D. Fedoseyenko,
2007, 129, 6887 – 6895; d) K. Haslinger, M. Peschke, C. Brieke, B. Philmus, L. E. Cooper, Y. Liu, I. Jhulki, S. E. Ealick, T. P.
E. Maximowitsch, M. J. Cryle, Nature 2015, 521, 105 – 109; Begley, J. Biol. Chem. 2015, 290, 3980 – 3986.
e) C. C. Forneris, M. R. Seyedsayamdost, Angew. Chem. Int. [105] a) D. W. Muldera, E. S. Boyd, R. Sarma, R. K. Lange, J. A.
Ed. 2018, 57, 8048 – 8052; Angew. Chem. 2018, 130, 8180 – 8184. Endrizzi, J. B. Broderick, J. W. Peters, Nature 2010, 465, 248 –
[83] J. M. Grandner, R. A. Cacho, Y. Tang, K. N. Houk, ACS Catal. 251; b) A. S. Byer, E. M. Shepard, J. W. Peters, J. B. Broderick,
2016, 6, 4506 – 4511. J. Biol. Chem. 2015, 290, 3987 – 3994; c) D. L. Suess, J. M.
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü
Kuchenreuther, L. De La Paz, J. R. Swartz, R. D. Britt, Inorg. [108] a) H. Nakamura, H. A. Hamer, G. Sirasani, E. P. Balskus, J.
Chem. 2016, 55, 478 – 487. Am. Chem. Soc. 2012, 134, 18518 – 18521; b) H. Nakamura,
[106] a) A. B. Smith, S. A. Kozmin, C. M. Adams, D. V. Paone, J. Am. E. E. Schultz, E. P. Balskus, Nat. Chem. Biol. 2017, 13, 916 –
Chem. Soc. 2000, 122, 4984 – 4985; b) A. B. Smith, C. M. 921.
Adams, S. A. Kozmin, D. V. Paone, J. Am. Chem. Soc. 2001,
123, 5357 – 5359. Manuscript received: July 10, 2018
[107] B. S. Moore, J. L. Chen, G. M. L. Patterson, R. E. Moore, Accepted manuscript online: August 28, 2018
Tetrahedron 1992, 48, 3001 – 3006. Version of record online: && &&, &&&&
Angew. Chem. Int. Ed. 2019, 58, 2 – 36 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü
Reviews
Biosynthesis Fantastic four: Generally, enzymes are
highly selective catalysts for single reac-
C. T. Walsh, B. S. Moore* &&&&—&&&& tions. However, some enzymes instead
control a series of reactions in a cascade-
Enzymatic Cascade Reactions in like fashion. This Review highlights four
Biosynthesis types of enzymatic cascade strategies,
mediated by nucleophilic, electrophilic,
pericyclic, and radical-based reactions,
observed in the biosynthesis of complex
natural products
&&&& www.angewandte.org 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2019, 58, 2 – 36
Ü
Ü