scholarly journals Engineering Fungal Nonreducing Polyketide Synthase by Heterologous Expression and Domain Swapping

2013 ◽  
Vol 15 (4) ◽  
pp. 756-759 ◽  
Author(s):  
Hsu-Hua Yeh ◽  
Shu-Lin Chang ◽  
Yi-Ming Chiang ◽  
Kenneth S. Bruno ◽  
Berl R. Oakley ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Polyketide Synthase ◽  
Domain Swapping

scholarly journals Initiation of Polyene Macrolide Biosynthesis: Interplay between Polyketide Synthase Domains and Modules as Revealed via Domain Swapping, Mutagenesis, and Heterologous Complementation

2011 ◽  
Vol 77 (19) ◽  
pp. 6982-6990 ◽  
Author(s):  
Sondre Heia ◽  
Sven E. F. Borgos ◽  
Håvard Sletta ◽  
Leticia Escudero ◽  
Elena M. Seco ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
Fungal Infections ◽  
Domain Swapping ◽  
Site Directed Mutagenesis ◽  
Active Site Residue ◽  
Polyene Macrolide ◽  
Content Type ◽  
Streptomyces Noursei ◽  
Strict Specificity ◽  
Different Levels

ABSTRACTPolyene macrolides are important antibiotics used to treat fungal infections in humans. In this work, acyltransferase (AT) domain swaps, mutagenesis, and cross-complementation with heterologous polyketide synthase domain (PKS) loading modules were performed in order to facilitate production of new analogues of the polyene macrolide nystatin. Replacement of AT0in the nystatin PKS loading module NysA with the propionate-specific AT1from the nystatin PKS NysB, construction of hybrids between NysA and the loading module of rimocidin PKS RimA, and stepwise exchange of specific amino acids in the AT0domain by site-directed mutagenesis were accomplished. However, none of the NysA mutants constructed was able to initiate production of new nystatin analogues. Nevertheless, many NysA mutants and hybrids were functional, providing for different levels of nystatin biosynthesis. An interplay between certain residues in AT0and an active site residue in the ketosynthase (KS)-like domain of NysA in initiation of nystatin biosynthesis was revealed. Some hybrids between the NysA and RimA loading modules carrying the NysA AT0domain were able to prime rimocidin PKS with both acetate and butyrate units upon complementation of arimA-deficient mutant of the rimocidin/CE-108 producerStreptomyces diastaticus. Expression of the PimS0 loading module from the pimaricin producer in the same host, however, resulted in production of CE-108 only. Taken together, these data indicate relaxed substrate specificity of NysA AT0domain, which is counteracted by a strict specificity of the first extender module KS domain in the nystatin PKS ofStreptomyces noursei.

scholarly journals A hybrid modular polyketide synthase obtained by domain swapping

1996 ◽  
Vol 3 (10) ◽  
pp. 833-839 ◽  
Author(s):  
Markiyan Oliynyk ◽  
Murray J.B. Brown ◽  
Jesús Cortés ◽  
James Staunton ◽  
Peter F. Leadlay
Keyword(s):  
Polyketide Synthase ◽  
Domain Swapping ◽  
Modular Polyketide Synthase

scholarly journals Discovery and biosynthesis of clostyrylpyrones from the obligate anaerobe Clostridium roseum

2020 ◽  
Author(s):  
Jeffrey S Li ◽  
Yongle Du ◽  
Di Gu ◽  
Wenlong Cai ◽  
Allison Green ◽  
...  
Keyword(s):  
Natural Products ◽  
Heterologous Expression ◽  
Natural Product ◽  
Biochemical Analysis ◽  
Polyketide Synthase ◽  
Gene Knockout ◽  
Anaerobic Bacteria ◽  
Obligate Anaerobe ◽  
New Family ◽  
Expression Studies

ABSTRACTAnaerobic bacteria are a promising new source for natural product discovery. Examination of extracts from the obligate anaerobe Clostridium roseum led to discovery of a new family of natural products, the clostyrylpyrones. The polyketide synthase-based biosynthetic mechanism of clostyrylpyrones is further proposed based on bioinformatic, gene knockout, biochemical analysis and heterologous expression studies.

scholarly journals Loseolamycins: A Group of New Bioactive Alkylresorcinols Produced after Heterologous Expression of a Type III PKS from Micromonospora endolithica

Molecules ◽  
2020 ◽  
Vol 25 (20) ◽  
pp. 4594
Author(s):  
Constanze Lasch ◽  
Nils Gummerlich ◽  
Maksym Myronovskyi ◽  
Anja Palusczak ◽  
Josef Zapp ◽  
...  
Keyword(s):  
Natural Products ◽  
Heterologous Expression ◽  
Transcriptional Activation ◽  
Polyketide Synthase ◽  
Biologically Active ◽  
Aliphatic Chain ◽  
Biosynthetic Pathways ◽  
Type Iii Polyketide Synthase ◽  
Type Iii ◽  
Potential Applications

Natural products are a valuable source of biologically active compounds with potential applications in medicine and agriculture. Unprecedented scaffold diversity of natural products and biocatalysts from their biosynthetic pathways are of fundamental importance. Heterologous expression and refactoring of natural product biosynthetic pathways are generally regarded as a promising approach to discover new secondary metabolites of microbial origin. Here, we present the identification of a new group of alkylresorcinols after transcriptional activation and heterologous expression of the type III polyketide synthase of Micromonospora endolithica. The most abundant compounds loseolamycins A1 and A2 have been purified and their structures were elucidated by NMR. Loseolamycins contain an unusual branched hydroxylated aliphatic chain which is provided by the host metabolism and is incorporated as a starter fatty acid unit. The isolated loseolamycins show activity against gram-positive bacteria and inhibit the growth of the monocot weed Agrostis stolonifera in a germination assay. The biosynthetic pathway leading to the production of loseolamycins is proposed in this paper.

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