Actinoplanes teichomyceticus ATCC 31121 as a cell factory for producing teicoplanin

Abstract

BackgroundTeicoplanin is a glycopeptide antibiotic used clinically in Europe and in Japan for the treatment of multi-resistant Gram-positive infections. It is produced by fermenting Actinoplanes teichomyceticus. The pharmaceutically active principle is teicoplanin A2, a complex of compounds designated T-A2-1-A2-5 differing in the length and branching of the fatty acid moiety linked to the glucosamine residue on the heptapeptide scaffold. According to European and Japanese Pharmacopoeia, components of the drug must be reproduced in fixed amounts to be authorized for clinical use.ResultsWe report our studies on optimizing the fermentation process to produce teicoplanin A2 in A. teichomyceticus ATCC 31121. Robustness of the process was assessed on scales from a miniaturized deep-well microtiter system to flasks and 3-L bioreactor fermenters. The production of individual factors T-A2-1-A2-5 was modulated by adding suitable precursors to the cultivation medium. Specific production of T-A2-1, characterized by a linear C10:1 acyl moiety, is enhanced by adding methyl linoleate, trilinoleate, and crude oils such as corn and cottonseed oils. Accumulation of T-A2-3, characterized by a linear C10:0 acyl chain, is stimulated by adding methyl oleate, trioleate, and oils such as olive and lard oils. Percentages of T-A2-2, T-A2-4, and, T-A2-5 bearing the iso-C10:0, anteiso-C11:0, and iso-C11:0 acyl moieties, respectively, are significantly increased by adding precursor amino acids L-valine, L-isoleucine, and L-leucine. Along with the stimulatory effect on specific complex components, fatty acid esters, oils, and amino acids (with the exception of L-valine) inhibit total antibiotic productivity overall. By adding industrial oils to medium containing L-valine the total production is comparable, giving unusual complex compositions.ConclusionsSince the cost and the quality of teicoplanin production depend mainly on the fermentation process, we developed a robust and scalable fermentation process by using an industrial medium in which a complex composition can be modulated by the combined addition of suitable precursors. This work was performed in the wild-type strain ATCC 31121, which has a clear genetic background. This is important for starting a rational improvement program and also helps to better control teicoplanin production during process and strain development.

Relationship between Glycopeptide Production and Resistance in the Actinomycete Nonomuraea sp. ATCC 39727

“…Batch fermentations were conducted using the industrial production medium FM2 (8 g/liter Bacto-yeast extract . Fermentation runs were started by adding a 10% (vol/vol) inoculum from the vegetative cultures into FM2 medium in a 2-literworking-volume P-100 Applikon glass reactor (height, 25 cm; diameter, 13 cm) equipped with an AD1030 Biocontroller and AD1032 motor (21,37). The temperature was maintained at 30°C, stirring was at 500 rpm (corresponding to 1.17 m/s of tip speed), and the aeration rate was 2 liters/min.…”

A multi-centre, phase IV study to evaluate the steady-state plasma concentration and serum bactericidal activity of a generic teicoplanin preparation

“…3 Teicoplanin is a glycopeptide antibiotic, a class which includes vancomycin, telavancin, bleomycin and ramoplanin. The pharmacologically active ingredient, teicoplanin, refers to a group of structurally similar chemical entities, termed teicoplanin A 2-1 to A [2][3][4][5] , which are microbial secondary metabolites produced by a number of non-ribosomal peptide synthetases to form a cyclic heptapeptide structure. The A 2-1 to A 2-5 structural forms account for 89-95% of active teicoplanin.…”

“…The A 2-1 to A 2-5 structural forms account for 89-95% of active teicoplanin. 4 The different chemical entities in the teicoplanin A 2 group have identical chemical core structures, and differ only in a variable glucosamine N-acyl fatty acid side chain ranging in length from 9-12 carbons, with or without branching. The identical teicoplanin core consists of a seven amino acid, tetracyclic peptide backbone, with three sugar residues.…”

16S rRNA-based Metagenomic Analysis of Endophytic Actinomycetes Diversity from Tinospora crispa L. Miers

“…As endophytic actinomycetes, Amycolatopsis and Actinoplanes were isolated from uncultivatedly rice paddy roots (Tian et al 2007). Amycolatopsis was found to have antibacterial and anti-fungal activity (Qin et al 2009), whereas Actinoplanes produced Teicoplanin, a glycopeptide antibiotic for the treatment of multiresistant gram-positive infection (Taurino et al 2011) and α-glucosidase inhibitors (Zhang et al 2003). Meanwhile the α-glucosidase inhibitors can also be produced by Streptomyces isolated from T. crispa (Pujiyanto et al 2012).…”