Biochemical and Genetic Insights into Asukamycin Biosynthesis

Rui, Zhe; Petříčková, Kateřina; Škanta, František; Pospı́šil, Stanislav; Yang, Yanling; Chen, Chung-Yung; Tsai, S.-C.; Floss, Heinz G.; Petřı́ček, Miroslav; Yu, Tianyi

doi:10.1074/jbc.m110.128850

Cited by 56 publications

(80 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…4). Double-FabH homologues may be employed to incorporate unusual starter units into final products in some biosynthetic pathways, for instance, AsuC3/C4 in the asukamycin production pathway (51). Substrate selectivity and stereochemistry of LstAB and LstC enzymology will be subjects of further research.…”

Section: Discussionmentioning

confidence: 99%

Operon for Biosynthesis of Lipstatin, the Beta-Lactone Inhibitor of Human Pancreatic Lipase

Bai

Zhang

Lin

et al. 2014

Appl Environ Microbiol

View full text Add to dashboard Cite

c Lipstatin, isolated from Streptomyces toxytricini as a potent and selective inhibitor of human pancreatic lipase, is a precursor for tetrahydrolipstatin (also known as orlistat, Xenical, and Alli), the only FDA-approved antiobesity medication for long-term use. Lipstatin features a 2-hexyl-3,5-dihydroxy-7,10-hexadecadienoic-␤-lactone structure with an N-formyl-L-leucine group attached as an ester to the 5-hydroxy group. It has been suggested that the ␣-branched 3,5-dihydroxy fatty acid ␤-lactone moiety of lipstatin in S. toxytricini is derived from Claisen condensation between two fatty acid substrates, which are derived from incomplete oxidative degradation of linoleic acid based on feeding experiments. In this study, we identified a six-gene operon (lst) that was essential for the biosynthesis of lipstatin by large-deletion, complementation, and single-gene knockout experiments. lstA, lstB, and lstC, which encode two ␤-ketoacyl-acyl carrier protein synthase III homologues and an acyl coenzyme A (acyl-CoA) synthetase homologue, were indicated to be responsible for the generation of the ␣-branched 3,5-dihydroxy fatty acid backbone. Subsequently, the nonribosomal peptide synthetase (NRPS) gene lstE and the putative formyltransferase gene lstF were involved in decoration of the ␣-branched 3,5-dihydroxy fatty acid chain with an N-formylated leucine residue. Finally, the 3␤-hydroxysteroid dehydrogenase-homologous gene lstD might be responsible for the reduction of the ␤-keto group of the biosynthetic intermediate, thereby facilitating the formation of the unique ␤-lactone ring.

show abstract

Section: Discussionmentioning

confidence: 99%

Operon for Biosynthesis of Lipstatin, the Beta-Lactone Inhibitor of Human Pancreatic Lipase

Bai

Zhang

Lin

et al. 2014

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…AsuC15 thioesterase is also proposed to suppress excess ω-cyclohexyl fatty acid formation and help to maintain membrane homeostasis by releasing the CHC residue from FAS ACP, as the percentage of ω-cyclohexyl fatty acids was increased fivefold in the asuC15 mutant (Rui et al 2010). The double role of AsuC15 TEII is an interesting example of primary and secondary metabolism interconnection.…”

Section: Functions Of Teiismentioning

confidence: 99%

Roles of type II thioesterases and their application for secondary metabolite yield improvement

Kotowska

Pawlik

2014

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

A large number of antibiotics and other industrially important microbial secondary metabolites are synthesized by polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs). These multienzymatic complexes provide an enormous flexibility in formation of diverse chemical structures from simple substrates, such as carboxylic acids and amino acids. Modular PKSs and NRPSs, often referred to as megasynthases, have brought about a special interest due to the colinearity between enzymatic domains in the proteins working as an “assembly line” and the chain elongation and modification steps. Extensive efforts toward modified compound biosynthesis by changing organization of PKS and NRPS domains in a combinatorial manner laid good grounds for rational design of new structures and their controllable biosynthesis as proposed by the synthetic biology approach. Despite undeniable progress made in this field, the yield of such “unnatural” natural products is often not satisfactory. Here, we focus on type II thioesterases (TEIIs)—discrete hydrolytic enzymes often encoded within PKS and NRPS gene clusters which can be used to enhance product yield. We review diverse roles of TEIIs (removal of aberrant residues blocking the megasynthase, participation in substrate selection, intermediate, and product release) and discuss their application in new biosynthetic systems utilizing PKS and NRPS parts.

show abstract

“…176 The activity of the putative flavin-dependent epoxidase (AsuE3) was verified by gene disruption experiments, which led to the accumulation of 4- hydroxyprotoasukamycin D ( 135 ) as the major product. 176 When compound 135 was fed to an S. lividans strain expressing the asuE3 gene, asukamycin A1 production was restored. Although AsuE3 has not been purified and biochemically characterized, its assignment as a monooxygenase is consistent with earlier 18 O 2 feeding studies of structurally related manumycin compounds, which demonstrated that the epoxide oxygen atom in this group of natural products is derived from O 2 .…”

Section: Epoxide Biosynthesismentioning

confidence: 99%