Speed dating for enzymes! Finding the perfect phosphopantetheinyl transferase partner for your polyketide synthase

AbstractThe biosynthetic pathways for the fungal polyketides bikaverin and bostrycoidin, from Fusarium verticillioides and Fusarium solani respectively, were reconstructed and heterologously expressed in S. cerevisiae alongside seven different phosphopantetheinyl transferases (PPTases) from a variety of origins spanning bacterial, yeast and fungal origins. In order to gauge the efficiency of the interaction between the ACP-domains of the polyketide synthases (PKS) and PPTases, each were co-expressed individually and the resulting production of target polyketides were determined after 48 h of growth. In co-expression with both biosynthetic pathways, the PPTase from Fusarium verticillioides (FvPPT1) proved most efficient at producing both bikaverin and bostrycoidin, at 1.4 mg/L and 5.9 mg/L respectively. Furthermore, the remaining PPTases showed the ability to interact with both PKS’s, except for a single PKS-PPTase combination. The results indicate that it is possible to boost the production of a target polyketide, simply by utilizing a more optimal PPTase partner, instead of the commonly used PPTases; NpgA, Gsp and Sfp, from Aspergillus nidulans, Brevibacillus brevis and Bacillus subtilis respectively.

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Phylogenomic analysis of polyketide synthase genes in actinomycetes: structural analysis of KS domains and modules of polyketide synthases

International Journal of Computational Biology and Drug Design ◽

10.1504/ijcbdd.2012.048281 ◽

2012 ◽

Vol 5 (2) ◽

pp. 89

Author(s):

Samreen Sarwar ◽

Mehboob Ahmed ◽

Shahida Hasnain

Keyword(s):

Structural Analysis ◽

Polyketide Synthase ◽

Polyketide Synthases ◽

Phylogenomic Analysis

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EFFICIENCY OF TRICHODERMA VIRIDE AND BACILLUS SUBTILIS AS BIOCONTROL AGENTS AGAINST ROOT ROT CAUSED BY FUSARIUM SOLANI IN TOMATO

Egyptian Journal of Agricultural Research ◽

10.21608/ejar.2019.111021 ◽

2019 ◽

Vol 97 (2) ◽

pp. 507-516

Author(s):

Mohamed KHALIL

Keyword(s):

Bacillus Subtilis ◽

Fusarium Solani ◽

Root Rot ◽

Trichoderma Viride ◽

Biocontrol Agents

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Engineered Biosynthesis of a Novel Amidated Polyketide, Using the Malonamyl-Specific Initiation Module from the Oxytetracycline Polyketide Synthase

Applied and Environmental Microbiology ◽

10.1128/aem.72.4.2573-2580.2006 ◽

2006 ◽

Vol 72 (4) ◽

pp. 2573-2580 ◽

Cited By ~ 83

Author(s):

Wenjun Zhang ◽

Brian D. Ames ◽

Shiou-Chuan Tsai ◽

Yi Tang

Keyword(s):

Polyketide Synthase ◽

Major Product ◽

Polyketide Synthases ◽

Type Ii ◽

Streptomyces Rimosus ◽

Aromatic Polyketides ◽

Starter Unit ◽

Necessary And Sufficient ◽

Bacterial Type

ABSTRACT Tetracyclines are aromatic polyketides biosynthesized by bacterial type II polyketide synthases (PKSs). Understanding the biochemistry of tetracycline PKSs is an important step toward the rational and combinatorial manipulation of tetracycline biosynthesis. To this end, we have sequenced the gene cluster of oxytetracycline (oxy and otc genes) PKS genes from Streptomyces rimosus. Sequence analysis revealed a total of 21 genes between the otrA and otrB resistance genes. We hypothesized that an amidotransferase, OxyD, synthesizes the malonamate starter unit that is a universal building block for tetracycline compounds. In vivo reconstitution using strain CH999 revealed that the minimal PKS and OxyD are necessary and sufficient for the biosynthesis of amidated polyketides. A novel alkaloid (WJ35, or compound 2) was synthesized as the major product when the oxy-encoded minimal PKS, the C-9 ketoreductase (OxyJ), and OxyD were coexpressed in CH999. WJ35 is an isoquinolone compound derived from an amidated decaketide backbone and cyclized with novel regioselectivity. The expression of OxyD with a heterologous minimal PKS did not afford similarly amidated polyketides, suggesting that the oxy-encoded minimal PKS possesses novel starter unit specificity.

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The mtaA Gene of the Myxothiazol Biosynthetic Gene Cluster from Stigmatella aurantiaca DW4/3-1 Encodes a Phosphopantetheinyl Transferase that Activates Polyketide Synthases and Polypeptide Synthetases

The Journal of Biochemistry ◽

10.1093/oxfordjournals.jbchem.a002821 ◽

2001 ◽

Vol 129 (1) ◽

pp. 119-124 ◽

Cited By ~ 40

Author(s):

N. Gaitatzis ◽

A. Hans ◽

R. Muller ◽

S. Beyer

Keyword(s):

Gene Cluster ◽

Biosynthetic Gene Cluster ◽

Polyketide Synthases ◽

Biosynthetic Gene ◽

Phosphopantetheinyl Transferase ◽

Stigmatella Aurantiaca

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The Fusarium verticillioides FUM Gene Cluster Encodes a Zn(II)2Cys6 Protein That Affects FUM Gene Expression and Fumonisin Production

Eukaryotic Cell ◽

10.1128/ec.00400-06 ◽

2007 ◽

Vol 6 (7) ◽

pp. 1210-1218 ◽

Cited By ~ 137

Author(s):

Daren W. Brown ◽

Robert A. E. Butchko ◽

Mark Busman ◽

Robert H. Proctor

Keyword(s):

Gene Cluster ◽

Polyketide Synthase ◽

Fusarium Verticillioides ◽

Gene Clusters ◽

Biosynthetic Gene Cluster ◽

Regulatory Proteins ◽

Wild Type ◽

Polyketide Synthase Gene ◽

Splice Forms ◽

Self Protection

ABSTRACT Fumonisins are mycotoxins produced by some Fusarium species and can contaminate maize or maize products. Ingestion of fumonisins is associated with diseases, including cancer and neural tube defects, in humans and animals. In fungi, genes involved in the synthesis of mycotoxins and other secondary metabolites are often located adjacent to each other in gene clusters. Such genes can encode structural enzymes, regulatory proteins, and/or proteins that provide self-protection. The fumonisin biosynthetic gene cluster includes 16 genes, none of which appear to play a role in regulation. In this study, we identified a previously undescribed gene (FUM21) located adjacent to the fumonisin polyketide synthase gene, FUM1. The presence of a Zn(II)2Cys6 DNA-binding domain in the predicted protein suggested that FUM21 was involved in transcriptional regulation. FUM21 deletion (Δfum21) mutants produce little to no fumonisin in cracked maize cultures but some FUM1 and FUM8 transcripts in a liquid GYAM medium. Complementation of a Δfum21 mutant with a wild-type copy of the gene restored fumonisin production. Analysis of FUM21 cDNAs identified four alternative splice forms (ASFs), and microarray analysis indicated the ASFs were differentially expressed. Based on these data, we present a model for how FUM21 ASFs may regulate fumonisin biosynthesis.

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Acyl Transferase Domains of Putative Polyketide Synthase (PKS) Genes in Aspergillus and Penicillium Producers of Ochratoxin A and the Evaluation of PCR Primers to Amplify PKS Sequences in Black Aspergillus Species

Food Science and Technology International ◽

10.1177/1082013208102743 ◽

2009 ◽

Vol 15 (1) ◽

pp. 97-105 ◽

Cited By ~ 3

Author(s):

P.V. Martínez Culebras ◽

A. Crespo-Sempere ◽

J.V. Gil ◽

D. Ramón

Keyword(s):

Secondary Metabolites ◽

Ochratoxin A ◽

Polyketide Synthase ◽

Pcr Primers ◽

Polyketide Synthases ◽

Aspergillus Species ◽

Acyl Transferase ◽

General Applicability ◽

Phylogenetic Methods ◽

Mycotoxin Biosynthesis

Fungal polyketide synthases (PKS) are responsible for the biosynthesis of several mycotoxins and other secondary metabolites. PKS genes in ochratoxin producing species from Aspergillus and Penicillum genera have been identified using a degenerate primer pair developed for the acyl transferase (AT) domain of fungal PKSs. Sequences of AT domains were aligned and analyzed using phylogenetic methods. The AT domain sequences appeared to be specific for a particular type of fungal PKSs and were related to PKSs involved in different mycotoxin biosynthesis pathways, including ochratoxin A. We have also developed primers suitable for amplifying AT domain sequences in strains belonging to the A. niger aggregate. DNA from most of the black Aspergillus species currently recognized was tested. Primers showed general applicability and other Aspergillus species belonging to section Nigri were successfully amplified.

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