ILI-8 Progress towards the heterologous expression of myxobacterial natural product assembly lines(Isolation and Structure Elucidation of Natural Products)

(1)

Symposium on the chemistry of natural products

NII-Electronic Library Service

Symposium on thechemistry of natural _products

IL

l-8

Progress towards the

heterologous

expression of myxobacterial natural

product

assembly

lines

RolfMttller

SaarlandUniversity,RO. Box 15l150,66041SaarbrUcken,Gerrnany

rom@mx.uni-saarland.de

Natural_productsof microbial origin are often biosynthesizedby multifunctionat

megasynthetases whose _genetic engineering and heterolegousexpression offers

considerable _promise,especially ifthenatural hostsare _geneticallydifficulttohandle, slow _growing, unculturable er even Lmknown

(1,2).

We have describeda

straightforward strategy that combines the _power of advanced DNA engineering

(recombiogenic

cloning) inEscherichiacoli with theutility of _pseudomonadsas the

heterologoushostfbrtheanalysis and mutagenesis _of known _and _unknown _secondary metabolite _pathways

(2,3).

As a

first

example, themyxochromide S biosynthetic_gene

cluster fromstigmatellaaurantiaca was rebuilt and engineered inE.coli to contain the

elements required forexpression in _{pseudomonads,}The _successfu1 _productionin

Pseudomonas _putido,at unprecedented levels,demonstratesthe feasibilityof a new

approach tothe analysis and mutagenesis of theseimportant_pathways

(4).

Recently,

R _puticlawas also used to express secondary metabolic _genes_of _unknown function

resulting inthe_productionofa novel compound frommyxobacteria

(5).

Inthe_{presentation,}

further

developments

such as metabolic _pathwayengineering of

Rputiclato enable _productionof _precursorsrequired

for

secondary metabolite

biosynthesiswill bedescribed.Inaddition, applications ofthe Red/ET technologyand

datarelated tothe use of otherheterologoushostswillbe_presented. References:

(1)Bode,H,B,,and MOIIer,R.,Angew, Chem.Int.Ed,,44:6828-46

(2)Wenzel,S.C,and MUIIer,R.,Curr.Opin.Biotechnol,.16:594-606

(3)Gross,F,,Gottschalk,D. and Mil11er,R,(2005)AppLMicrobioLBiotechnol.,68,66-74 .

(4)Wenzel,S.C.,Gross,F.,Zhang,Y.,Fu,J.,Stewart,F,,and Mti11er,R.(2005)Chem. Biol.,12,349-356

(5)Gross,F.,Luniak,N.,Perleva.O,,Gaitatzjs,N.,Jenke-Kodama,H.,Gerth,K.,Gottscha]k,D.,Dittmann,E,and Muller,R.,₍₂₀₀₆₎Arch Microbiol,185,28-38