Biosynthetic Origin of Hydrogen Atoms in the Lipase Inhibitor Lipstatin

Goese, Markus; Eisenreich, Wolfgang; Kupfer, Ernst; Bacher, Adelbert

doi:10.1074/jbc.m003094200

Cited by 16 publications

(27 citation statements)

References 14 publications

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“…One substrate, 3-hydroxytetradeca-5,8-dienoyl-CoA, is directly obtained from ␤-oxidation. However, in analogy to the initiation of fatty acid synthesis, activation and carboxylation of octanoic acid are needed to form hexylmalonyl-CoA to serve as the second substrate (17)(18)(19)(20)(21). LstA, LstB, LstC, and a few enzymes from primary metabolism are plausibly involved in these substrate activation and condensation steps.…”

Section: Discussionmentioning

confidence: 99%

“…According to lipstatin biosynthesis models established on the basis of early feeding experiments with S. toxytricini, the first committed step is Claisen condensation between (3S,5Z,8Z)-3-hydroxytetradeca-5,8-dienoyl-CoA and activated octanoic acid to afford the 22-carbon ␣-branched fatty acid backbone (17)(18)(19)(20)(21). Both substrates of Claisen condensation are obtained from incomplete degradation of linoleic acid.…”

Section: Discussionmentioning

confidence: 99%

“…The ␣-branched fatty acid moiety of lipstatin is derived from Claisen condensation between octanoyl coenzyme A (octanoyl-CoA) and 3-hydroxytetradeca-5,8-dienoyl-CoA, both of which are obtained from incomplete ␤-oxidation of linoleic acid (17)(18)(19)(20)(21). In addition, the H-2 atom of the octanoic acid precursor is displaced with the solvent proton in the final product lipstatin (20).…”

mentioning

confidence: 99%

“…The biosynthesis of lipstatin in S. toxytricini has been intensively studied based on 15 N, 13 C, and deuterium labeling experiments by the Bacher group (17)(18)(19)(20)(21). The ␣-branched fatty acid moiety of lipstatin is derived from Claisen condensation between octanoyl coenzyme A (octanoyl-CoA) and 3-hydroxytetradeca-5,8-dienoyl-CoA, both of which are obtained from incomplete ␤-oxidation of linoleic acid (17)(18)(19)(20)(21).…”

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confidence: 99%

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

Bai

Zhang

Lin

et al. 2014

Appl Environ Microbiol

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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.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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

Bai

Zhang

Lin

et al. 2014

Appl Environ Microbiol

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“…Early studies have elucidated that the carbon skeleton of lipstatin molecule is biosynthesized via Claisen condensation of two fatty acid precursors, the 8-carbon atoms (octanoic acid), and 14-carbon atoms (tetra deca-5, 8-dienoic acid) (Eisenreich et al, 1997;Goese et al, 2000Goese et al, , 2001. As per this biosynthetic pathway and the basic building unit of lipstatin, oils rich in fatty acids are good resources for precursors.…”

Section: Full Papermentioning

confidence: 99%

Enhanced production of lipstatin from Streptomyces toxytricini by optimizing fermentation conditions and medium

Zhu

Wang

et al. 2014

J. Gen. Appl. Microbiol.

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This paper is concerned with optimization of fermentation conditions for lipstatin production with. Under these conditions, a maximum lipstatin of 4.208 g/ml was achieved in verification experiments in 500 ml shake flasks. Key words: divalent cations; L-leucine; lipstatin; Streptomyces toxytricini IntroductionLipstatin is an irreversible inhibitor of pancreatic lipases produced by Streptomyces toxytricini Weibel et al., 1987). Lipstatin is of considerable importance as the key intermediate for the preparation of tetrahydrolipstatin (THL, Orlistat), which is a drug designed for the prophylaxis and treatment of diseases associated with obesity. Lipstatin contains a β-lactone structure that probably accounts for the irreversible lipase inhibition by covalent modification of the serine residue of its catalytic triad (Luthra et al., 2013b).The wide clinical application of orlistat has promoted the commercial production of lipstatin (Zohrabian, 2100). Hence, investigations into the improvement of the productions of lipstatin are of commercial importance. In an attempt to improve lipstatin production, improvement of lipstatinproducing strains and fermentation processes has been carried out during the past decade.S. toxytricini strains with an increased lipstatin fermentation unit have been achieved through modification and improvement by utilizing physical and chemical mutagenesis. A high lipstatin-producing strain SIPI-HJ-80 was obtained by ultraviolet ray and microwave treatment, which showed an increased productivity by 2.5 times over the original strain SIPI-UM-5 (Huang et al., 2006). More recently, a N-methyl-N -nitro-N-nitrosoguanidine (NTG) mutated strain of S. toxytricini (derived from strain ATCC 19813) was reported to have a lipstatin production of 2.88 mg/g at 264 h of fermentation time in a shake flask (Luthra et al., 2013a).With the strain S. toxytricini ATCC 19813, 8-fold enhancement (from 0.097 g/L to 0.885 g/L) in lipstatin production in a shake flask was obtained through medium optimization. It is also indicated that soya oil, soya lecithin and soya bean flour had significant effects on lipstatin production (Luthra et al., 2012). An increased lipstatin titer of 1.980 g/L was observed with selected S. toxytricini strain NRRL 15443 (Kumar et al., 2012). Further, a maximum lipstatin production of 3.290 g/L was achieved via a full factorial medium Full PaperEnhanced production of lipstatin from Streptomyces toxytricini by optimizing fermentation conditions and medium (Received February 28, 2014; Accepted April 3, 2014) None of the authors of this manuscript has any financial or personal relationship with other people or organizations that could inappropriately influence their work.

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Proteinreaktive Naturstoffe

2005

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In der Post‐Genom‐Ära steht man vor der gewaltigen Aufgabe, die Strukturen und Funktionen zehntausender codierter Proteine zuzuweisen. Zur Analyse der Proteinfunktion werden neue Technologien mit großer Bandbreite entwickelt, etwa die aktivitätsbasierte Proteinanalyse (activity‐based protein profiling, ABPP). Bei diesem Verfahren sollen auf das aktive Zentrum gerichtete chemische Sonden entwickelt werden, um die Enzyme in ganzen Proteomen zu analysieren. Derartige chemische Proteomtechnologien können von proteinreaktiven Naturstoffen lernen, die Enzyme der unterschiedlichsten Klassen durch ein bemerkenswert vielfältiges Spektrum an Mechanismen kovalent modifizieren und somit eine reiche Quelle an Konzepten für die Entwicklung gegen aktive Zentren gerichteter Proteomsonden bilden. Hier werden wir anhand unterschiedlicher Naturstoffe zeigen, wie ihre Wirkmechanismen zum Fortschritt chemischer Proteomtechnologien wie ABPP beigetragen haben.

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Biosynthetic Origin of Hydrogen Atoms in the Lipase Inhibitor Lipstatin

Cited by 16 publications

References 14 publications

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

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

Enhanced production of lipstatin from Streptomyces toxytricini by optimizing fermentation conditions and medium

Proteinreaktive Naturstoffe

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