Biosynthetic Precursors of the Lipase Inhibitor Lipstatin

Schuhr, Christoph A.; Eisenreich, Wolfgang; Goese, Markus; Stohler, Peter; Kupfer, Ernst; Bacher, Adelbert

doi:10.1021/jo016285v

Cited by 16 publications

(21 citation statements)

References 8 publications

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“…It is found in this study that the addition of short chain L-leucine was effective in the middle phase of fermentation when the synthesis of lipstatin started gradually, whereas the addition of long-chain octanoic acid was more effective in the late phase of fermentation when the synthesis of lipstatin is most active. Our results provide further evidence in support of precursors for biosynthesis of lipstatin as described by Schuhr et al (2002). As described in a patent (US 4598089, Hadvary et al, 1988), feeding of L-leucine or N-formyl-L-leucine during the fermentation process is identified as being particularly useful in order to increase the final yield of lipstatin with S. toxytricini NRRL15443.…”

Section: Effects Of Precursors On Lipstatin Productionsupporting

confidence: 82%

“…Linoleic acid contains the same number of double bonds as lipstatin chain and it is the basic building unit. Further experiments with feeding putative intermediates in the process of fermentation indicated that linoleic acid, caprylic acid and N-formyl-L-leucine or preferably L-leucine are effective precursors of lipstatin biosynthesis (Schuhr et al, 2002;Eisenreich et al, 2003;Demirev et al, 2010).…”

Section: Full Papermentioning

confidence: 99%

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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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Section: Effects Of Precursors On Lipstatin Productionsupporting

confidence: 82%

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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“…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).…”

mentioning

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).…”

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confidence: 99%

“…Subsequently, Pks13, a type I polyketide synthase (PKS) with multiple functional domains, takes the two substrates by attaching them to two phosphopantetheine-binding (PPB) domains and catalyzes Claisen condensation between two tethered substrates by the ketosynthase (KS) domain (23). Furthermore, when [ 13 C-formyl, 15 N]-N-formyl-leucine was fed to S. toxytricini fermentation broth, only the 15 N-leucine moiety of the double-labeled chemical was incorporated into lipstatin, suggesting that the incorporation of leucine takes place after hydrolysis of the formamide motif and that the formyl group is transferred to the L-leucine residue afterwards (21).…”

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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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Chemoenzymatic Synthesis of the HMG‐CoA Reductase Inhibitor Rosuvastatin and Natural Styryl Lactone Cryptomoscatone E1

et al. 2017

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Racemic syn‐ethyl (E)‐2‐(2,2‐dimethyl‐6‐styryl‐1,3‐dioxan‐4‐yl)acetate is hydrolyzed with high enantioselectivity by Novozyme‐435 in 0.05 m sodium phosphate buffer at room temperature. The optically pure unreacted ethyl 2‐((4R,6S)‐2,2‐dimethyl‐6‐((E)‐styryl)‐1,3‐dioxan‐4‐yl)acetate and hydrolyzed (4S,6R)‐acid are used in the synthesis of HMG‐CoA reductase inhibitor rosuvastatin and the naturally occurring styryl lactone cryptomoscatone E1, respectively.

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Biosynthetic Precursors of the Lipase Inhibitor Lipstatin

Cited by 16 publications

References 8 publications

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

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

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

Chemoenzymatic Synthesis of the HMG‐CoA Reductase Inhibitor Rosuvastatin and Natural Styryl Lactone Cryptomoscatone E1

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