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

Abstract

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.

Implication of mutagenesis and precursor supplementation towards the enhancement of lipstatin (an antiobesity agent) biosynthesis through submerged fermentation using Streptomyces toxytricini

“…Mutant strains of S. toxytricini showing highest production of lipstatin were used for precursor supplementation. Metabolic precursors such as citric acid (1 g/L), thiamine (1 g/L), and biotin (1 g/L) were supplemented after 96 h (idiophase) of incubation of fermentation medium (Zhu et al 2014;Kumar and Dubey 2016). In this study, one precursor was supplemented at one time and change in lipstatin production was analyzed.…”

“…Natural producers of lipstatin are Streptomyces toxytricini (S. toxytricini) and S. virginiae, but only S. toxytricini is explored for the production of lipstatin at commercial scale. To enhance the lipstatin production from S. toxytricini, researchers performed optimization of culture conditions and medium modifications which enhanced production of lipstatin to 3290 mg/L (Luthra et al 2013b), 4208 mg/L (Zhu et al 2014) and 2262.63 mg/L (Kumar and Dubey 2016). Moreover, it was also suggested that further improvement in lipstatin production may be performed by metabolic engineering and strain improvement approaches (Kumar and Dubey 2015).…”

“…To enhance the production of lipstatin by random mutagenesis of S. toxytricini; Luthra et al (2013a) have reported the 2.34 and 2.88 mg/g production of lipstatin through UV and NTG mutagenesis, respectively. Including this, Zhu et al (2014) have reported enhancement in lipstatin production through optimization of culture conditions and precursor supplementation to mutated strain of S. toxytricini.…”

Current trends and future prospects of lipstatin: a lipase inhibitor and pro-drug for obesity

“…Though, scientific data supports the enhancement in the lipstatin production upto 4208µg/ml 84 Researchers must explore metabolic engineering approach for strain development though it is dependent on the knowledge of metabolomics, genomics, transciptomics, flux investigation, and efficient genetic tools.…”

In Vivo Assay Reveals Microbial OleA Thiolases Initiating Hydrocarbon and β-Lactone Biosynthesis

“…For example, salinosporamide A, a ␤-lactone natural product, is currently in phase three clinical trials for the treatment of glioblastoma (7). Lipstatin, produced by various Streptomyces spp., is hydrogenated industrially to make the FDA-approved antiobesity drug tetrahydrolipstatin (8,9).…”