LovG: The Thioesterase Required for Dihydromonacolin L Release and Lovastatin Nonaketide Synthase Turnover in Lovastatin Biosynthesis

Xu, Wei; Chooi, Yit‐Heng; Choi, Jin Wook; Li, Shuang; Vederas, John C.; Silva, Nancy A. Da; Tang, Yi

doi:10.1002/anie.201302406

Cited by 110 publications

(86 citation statements)

References 22 publications

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“…The metabolite data were furthermore used to propose a model for fusarielin biosynthesis in F. graminearum (Figure 3) based on models for lovastatin biosynthesis [3,10,15]. Analyses of the fusarielin and lovastatin gene clusters showed that there is some sequence similarity for the PKS's (38% identity), trans-thioesterases (28% identity), and trans-ERs (46% identity), with E values from blastP analyses ranging from 0.0 to 1 × 10 −8 (Supplementary Table S1).…”

Section: Posmentioning

confidence: 99%

“…This suggest that there are several similarities to the biosynthetic pathway of lovastatin in which the polyketide chain is reduced at five sites by a trans-ER (lovC) [6,7] before it is released from the PKS by a trans-thioesterase (LovG) [10]. The Diels-Alder reaction is proposed to occur at the pentaketide stage in lovastatin biosynthesis, and it is therefore possible that fusarielin biosynthesis follows a similar route.…”

Section: Posmentioning

confidence: 99%

“…After decalin formation and subsequent chain elongation and modification, the resulting decalin polyketide product can be released from the PKS by various mechanisms. A terminal PKS reduction domain catalyzes release in the betaenone and equisetin biosynthesis pathway [8,9], while a trans-acting thioesterase (LovG) is involved in the lovastatin pathway [10].…”

Section: Introductionmentioning

confidence: 99%

“…A terminal PKS reduction domain catalyzes release in the betaenone and equisetin biosynthesis pathway [8,9], while a trans-acting thioesterase (LovG) is involved in the lovastatin pathway [10].…”

Section: Introductionmentioning

confidence: 99%

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Functional Analysis of the Fusarielin Biosynthetic Gene Cluster

et al. 2016

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Abstract:Fusarielins are polyketides with a decalin core produced by various species of Aspergillus and Fusarium. Although the responsible gene cluster has been identified, the biosynthetic pathway remains to be elucidated. In the present study, members of the gene cluster were deleted individually in a Fusarium graminearum strain overexpressing the local transcription factor. The results suggest that a trans-acting enoyl reductase (FSL5) assists the polyketide synthase FSL1 in biosynthesis of a polyketide product, which is released by hydrolysis by a trans-acting thioesterase (FSL2). Deletion of the epimerase (FSL3) resulted in accumulation of an unstable compound, which could be the released product. A novel compound, named prefusarielin, accumulated in the deletion mutant of the cytochrome P450 monooxygenase FSL4. Unlike the known fusarielins from Fusarium, this compound does not contain oxygenized decalin rings, suggesting that FSL4 is responsible for the oxygenation.

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Section: Posmentioning

confidence: 99%

Section: Posmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Functional Analysis of the Fusarielin Biosynthetic Gene Cluster

et al. 2016

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“…With the development of synthetic biology, reconstruction of biosynthetic pathways in chassis organisms has been proved to be a possible solution to these problems (Ro et al 2006;Galanie et al 2015). The biosynthetic gene cluster and catalytic role of each functional gene product are well characterized for lovastatin (Kennedy et al 1999;Xie et al 2006;Barriuso et al 2011;Xu et al 2013). Previously, we assembled the biosynthetic pathway of lovastatin in the methylotrophic yeast P. pastoris (Fig.…”

Section: Introductionmentioning

confidence: 99%

Improved methanol-derived lovastatin production through enhancement of the biosynthetic pathway and intracellular lovastatin efflux in methylotrophic yeast

Liu

Bai

et al. 2018

Bioresour. Bioprocess.

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Background: Methanol has attracted interest as a substrate for improvement of product titers and yields of bioprocesses, because methanol has a more reduced state than sugars do and can be renewably synthesized from abundant natural gas supplies. Our aim was to engineer methylotrophic Pichia pastoris to convert methanol into value-added lovastatin more productively. Results:A strengthened biosynthetic pathway of lovastatin was constructed through the assembly of three modules with increasing module-specific antibiotic stress in the methylotrophic yeast P. pastoris. The resulting strain (P. p/LV_V#9) produced 287.5 ± 2.0 mg/L lovastatin in a 5-L bioreactor from methanol. The production was further improved by identification and overexpression of a statin pump protein, TapA, a membrane protein capable of lovastatin efflux out of the cell. A TapA-overexpressing strain, P. p/LV_V#9-TapA, produced 419.0 ± 9.5 mg/L lovastatin from methanol: 46% more than P. p/LV_V#9 did, and 520% more relative to the strain (P. p/LV_SC) with single-copy genes. Conclusions:A methylotrophic yeast strain producing 419.0 ± 9.5 mg/L lovastatin was constructed by optimization of biosynthetic gene dosages and coexpression of a statin pump protein; these results proved that P. pastoris is a promising chassis organism for natural-product biosynthesis. A membrane protein, TapA was found to perform the function of exporting intracellular lovastatin and enhanced lovastatin production.

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Production of New Cladosporin Analogues by Reconstitution of the Polyketide Synthases Responsible for the Biosynthesis of this Antimalarial Agent

et al. 2015

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The antimalarial agent cladosporin is an anomolar inhibitor of the Plasmodium falciparum lysyl-tRNAs ynthetase,a nd exhibits activity against both blood-and liver-stage infection. Cladosporin can be isolated from the fungus Cladosporium cladosporioides,w here it is biosynthesized by ah ighly reducing (HR) and an on-reducing (NR) iterative type Ipolyketide synthase (PKS) pair.Genome sequencing of the host organism and subsequent heterologous expression of these enzymes in Saccharomyces cerevisiae produced cladosporin, confirming the identity of the putative gene cluster. Incorporation of ap entaketide intermediate analogue indicated a5 + +3a ssembly by the HR PKS Cla2 and the NR PKS Cla3 during cladosporin biosynthesis.A dvanced-intermediate analogues were synthesized and incorporated by Cla3 to furnish new cladosporin analogues.Aputative lysyl-tRNA synthetase resistance gene was identified in the cladosporin gene cluster.A nalysis of the active site emphasizes key structural features thought to be important in resistance to cladosporin.Cladosporin (1;a sperentin) is at ricyclic octaketide that is produced by several fungal species,i ncluding Cladosporium, [1][2][3][4] Chaetomium, [5] Penicillium, [6] Eurotium, [7,8] and Aspergillus.[9] Cladosporin exhibits interesting bioactivity, including antifungal, antibiotic,a nd plant-growth inhibitory properties and anti-inflammatory responses in mouse lung tissue.[10] Recently,cladosporin has been shown to be ananomolar inhibitor of Plasmodium falciparum blood-and liverstage proliferation.[11] Although many antimalarial agents currently exist, endoperoxides are the only class of molecules for which resistance has not significantly developed, and even these do not inhibit the asymptotic liver-stage infection. The bioactivity of cladosporin represents apromising lead for the treatment of malaria, and several studies on this topic have been published. [12,13] Previously,w ei nvestigated the biosynthesis of several related fungal polyketides that belong to the resorcylic acid lactone (RAL) and dihydroxyphenylacetic acid lactone (DAL) containing polyketides,i ncluding hypothemycin (RAL type), [14] radicicol (RAL type), [15] and dehydrocurvularin (DAL type). [16][17][18][19] Biosynthesis of these polyketides requires cooperative action of two iterative type Ipolyketide synthases (PKSs): ah ighly reducing (HR) PKS and an onreducing (NR) PKS.B ased on structural similarities,w e hypothesized that cladosporin is also biosynthesized by aHR and an NR PKS.O ur early work assigned the absolute stereochemistry of cladosporin. [2,3] More recently,t he total synthesis of cladosporin and its diastereomer,isocladosporin, has been reported. [20,21] To better understand PKS assembly and enable analogue production by synthetic biology,w e sought to heterologously express and reconstitute cladosporin expression in Saccharomyces cerevisiae.Hence,wesequenced the genome of the producer, Cladosporium cladosporioides UAMH 5063. This resulted in 30 Mb of genomic information over at otal of...

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LovG: The Thioesterase Required for Dihydromonacolin L Release and Lovastatin Nonaketide Synthase Turnover in Lovastatin Biosynthesis

Abstract: The cryptic thioesterase LovG is found to be responsible for product release from the lovastatin nonaketide synthase (LNKS or LovB). The same enzyme also helps improving turnover of LovB through hydrolysis of incorrectly tailored intermediates.

Cited by 110 publications

References 22 publications

Functional Analysis of the Fusarielin Biosynthetic Gene Cluster

Functional Analysis of the Fusarielin Biosynthetic Gene Cluster

Improved methanol-derived lovastatin production through enhancement of the biosynthetic pathway and intracellular lovastatin efflux in methylotrophic yeast

Production of New Cladosporin Analogues by Reconstitution of the Polyketide Synthases Responsible for the Biosynthesis of this Antimalarial Agent

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