Purification and N-Terminal Amino Acid Sequence of Solanapyrone Synthase, A Natural Diels-Alderase from<i>Alternaria solani</i>

Katayama, Kinya; Kobayashi, Tomonori; Chijimatsu, Masao; Ichihara, Akitami; Oikawa, Hideaki

doi:10.1271/bbb.70600

Cited by 16 publications

(17 citation statements)

References 6 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The sol5 gene encodes a polypeptide of 515 amino acid‐long polypeptide, which was considered to be SPS, an enzyme responsible for the Diels–Alder cycloaddition in solanapyrone biosynthesis. The presence in its N terminus of a signal peptide of 20 amino acids for secretion was predicted by the SignalIP server25 and the sequence following after the 25th amino acid was identical with that found in the partially purified SPS 19. A conserved domain search indicated the presence of a flavin‐binding domain (Pro95 to Ser230) in which the His128 residue was suggested as a site of covalent flavinylation (Figure S1 in the Supporting Information).…”

Section: Resultsmentioning

confidence: 80%

“…Although SPS was not fully purified, its oxidase activity could not be decoupled from the cycloaddition activity, and so SPS was considered to be an oxidase converting 2 into 3 , which is then cyclized and released as 1 18. Recently, Katayama et al reported that a more than 1630‐fold purification of SPS from a crude extract of A. solani was not sufficient to obtain a homogeneous SPS preparation, due to its instability and degradation by contaminating proteases 19…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Solanapyrone Synthase, a Possible Diels–Alderase and Iterative Type I Polyketide Synthase Encoded in a Biosynthetic Gene Cluster from Alternaria solani

et al. 2010

Self Cite

View full text Add to dashboard Cite

The solanapyrone biosynthetic gene cluster was cloned from Alternaria solani. It consists of six genes-sol1-6-coding for a polyketide synthase, an O-methyltransferase, a dehydrogenase, a transcription factor, a flavin-dependent oxidase, and cytochrome P450. The prosolanapyrone synthase (PSS) encoded by sol1 was expressed in Aspergillus oryzae and its product was identified as desmethylprosolanapyrone I (8). Although PSS is closely related to the PKSs/Diels-Alderases LovB and MlcA of lovastatin and compactin biosynthesis, it did not catalyze cycloaddition. Sol5, encoding a flavin-dependent oxidase (solanapyrone synthase, SPS), was expressed in Pichia pastoris and purified. The purified recombinant SPS showed activity for the formation of (-)-solanapyrone A (1) from achiral prosolanapyrone II (2), establishing that this single enzyme catalyzes both the oxidation and the subsequent cycloaddition reaction, possibly as a Diels-Alder enzyme.

show abstract

Section: Resultsmentioning

confidence: 80%

Section: Introductionmentioning

confidence: 99%

Solanapyrone Synthase, a Possible Diels–Alderase and Iterative Type I Polyketide Synthase Encoded in a Biosynthetic Gene Cluster from Alternaria solani

et al. 2010

Self Cite

View full text Add to dashboard Cite

show abstract

“…102,106 Sol5 (solanapyrone synthase) also appears to accelerate the intramolecular [4 + 2]-cycloaddition of 42 to produce a mixture of solanapyrone A ( 43 ) and solanapyrone D ( 45 ). 33,95,102,106,107 In the final step of the pathway, the dehydrogenase Sol3 reduces the aldehyde functionality of 43 and 45 to provide 44 and 46 , respectively. 102 …”

Section: Solanapyrone Synthasementioning

confidence: 99%

Natural [4 + 2]-Cyclases

et al. 2016

View full text Add to dashboard Cite

[4 + 2]-Cycloadditions are increasingly being recognized in the biosynthetic pathways of many structurally complex natural products. A relatively small collection of enzymes from these pathways have been demonstrated to increase rates of cyclization and impose stereochemical constraints on the reactions. While mechanistic investigation of these enzymes is just beginning, recent studies have provided new insights with implications for understanding their biosynthetic roles, mechanisms of catalysis and evolutionary origin.

show abstract

“…Using crude enzyme, we found that both 13 and 14 were converted into 15 and 16 [6]. Although purification of solanapyrone synthase (SPS) was hampered by its instability, it has been purified as a single band on SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) [26]. Recently, the gene cluster responsible for biosynthesis of solanapyrones has been identified by homology based PCR (polymerase chain reaction) and genome walking [27].…”

Section: Macrophomate Synthasementioning

confidence: 99%

The Diels–Alderase Never Ending Story

Minami

Oikawa

2011

Biomimetic Organic Synthesis

Self Cite

View full text Add to dashboard Cite

The biosynthesis of the cholesterol-lowering drug lovastatin isolated from Aspergillus terreus has been investigated extensively by Vederas and coworkers [10]. Incorporation experiments with multiple labeled acetate and 18 O-oxygen suggested that it is biosynthesized via the polyketide pathway. Based on feeding studies and co-occurrence of 4a,5-dihydromonacolin L (3), this compound was speculated as an intermediate. This was confirmed by the successful conversion of 3 into lovastatin, using a blocked mutant of A. terreus (Scheme 21.2) [11]. Because lovastatin does not have an electron-withdrawing group in the dienophile moiety, it was proposed that the requisite Diels-Alder reaction occurred at the hexaketide stage. However, all efforts to convert 13 C-labeled hexaketide precursor 1b into lovastatin were unsuccessful due to non-enzymatic cycloadditions affording a 1 : 1 mixture of undesired diastereomers 4b (endo) and 4c (exo) in aqueous media (half life of 1b: two days) (Scheme 21.2). This clearly showed significant rate acceleration for the cycloaddition of 1b in aqueous media [12].

show abstract

Purification and N-Terminal Amino Acid Sequence of Solanapyrone Synthase, A Natural Diels-Alderase fromAlternaria solani

Abstract: The first natural Diels-Alderase, solanapyrone synthase, was purified 1,630-fold from a crude extract. The 41-kDa protein on SDS-polyacrylamide gel electrophoresis was identified as truncated solanapyrone synthase, and its N-terminal amino acid sequence was found to be QETQNLNNFLESNAINP.

Cited by 16 publications

References 6 publications

Solanapyrone Synthase, a Possible Diels–Alderase and Iterative Type I Polyketide Synthase Encoded in a Biosynthetic Gene Cluster from Alternaria solani

Solanapyrone Synthase, a Possible Diels–Alderase and Iterative Type I Polyketide Synthase Encoded in a Biosynthetic Gene Cluster from Alternaria solani

Natural [4 + 2]-Cyclases

The Diels–Alderase Never Ending Story

Contact Info

Product

Resources

About