Enhancing effect of lysine combined with other compounds on cephamycin C production in Streptomyces clavuligerus

Abstract

BackgroundLysine plays an important role in Streptomyces clavuligerus metabolism; it takes part in its catabolism, via cadaverine, and in its secondary metabolism, in which lysine is converted via 1-piperideine-6-carboxylate to alpha-aminoadipic acid, a beta-lactam antibiotic precursor. The role of lysine as an enhancer of cephamycin C production, when added to production medium at concentrations above 50 mmol l-1, has already been reported in the literature, with some studies attributing a positive influence to multifunctional diamines, among other compounds. However, there is a lack of research on the combined effect of these compounds on antibiotic production.ResultsResults from experimental design-based tests were used to conduct response surface-based optimization studies in order to investigate the synergistic effect of combining lysine with cadaverine, putrescine, 1,3-diaminopropane, or alpha-aminoadipic acid on cephamycin C volumetric production. Lysine combined with cadaverine influenced production positively, but only at low lysine concentrations. On the whole, higher putrescine concentrations (0.4 g l-1) affected negatively cephamycin C volumetric production. In comparison to culture media containing only lysine as additive, combinations of this amino acid with alpha-aminoadipic acid or 1,3-diaminopropane increased cephamycin C production by more than 100%.ConclusionThis study demonstrated that different combinations of lysine with diamines or lysine with alpha-aminoadipic acid engender significant differences with respect to antibiotic volumetric production, with emphasis on the benefits observed for lysine combined with alpha-aminoadipic acid or 1,3-diaminopropane. This increase is explained by mathematical models and demonstrated by means of bioreactor cultivations. Moreover, it is consistent with the positive influence of these compounds on lysine conversion to alpha-aminoadipic acid, a limiting step in cephamycin C production.

Streptomyces clavuligerus strain selection for clavulanic acid biosynthesis: a study based on culture composition effects and statistical analysis

“…Using the NZ-Amine medium a CA production of 11.80 mg·L -1 was achieved. Likewise, the NZ-amine has lysine as the main amino acid source with a high concentration (430 mg·L -1 ), which favors cephamycin C biosynthesis [20]. S. clavuligerus grown in the chemically defined medium GSPA produced 25.50 mg CA·L -1 ; this medium contains proline and asparagine which facilitate microbial anabolic activity conducing to increased CA biosynthesis.…”

Homologous expression of lysA encoding diaminopimelic acid (DAP) decarboxylase reveals increased antibiotic production in Streptomyces clavuligerus

“…Fermentation optimization or metabolic engineering approaches, or both, were used to overproduce CephC in the cell. The addition of L-lysine, its product, i.e., α-AAA, or its precursor, i.e., diaminopimelate, stimulated the production of CephC in S. clavuligerus [2,11,12]. Similarly, supplementation of the fermentation medium with L-lysine or diamines putrescine, cadaverine, and diaminopropane also enhanced cephamycin production in Nocardia lactamdurans, another cephamycin producer actinomycete [13,14].…”

“…Of these secondary metabolites, CephC is a naturally occurring cephamycin antibiotic and potent against penicillinresistant bacteria via a methoxyl group at the C-7 position of the cephalosporin core. CephC is highly stable at neutral pH and used as a raw material to synthesize semisynthetic antibiotics such as cefoxitin, cefmetazole, temocillin, and cefotetan [1,2]. CA is a β-lactamase inhibitor used in combination with β-lactam antibiotics such as amoxicillin (Augmentin™) and ticarcillin (Timentin™) against β-lactamase-producing bacteria [3].…”

“…CA is a β-lactamase inhibitor used in combination with β-lactam antibiotics such as amoxicillin (Augmentin™) and ticarcillin (Timentin™) against β-lactamase-producing bacteria [3]. Non-β-lactam compounds such as holomycin with pyrrolidine structure and tunicamycin with a glucosaminecontaining structure are also produced by S. clavuligerus [2,4]. Tunicamycin is a fatty acyl nucleoside-type antibiotic and composed of a 14-membered gene cluster lacking clustersituated regulator (CSR) [5].…”

Bioprocesso De Produção Do Antitumoral Holomicina Por Streptomyces Clavuligerus

“…Dados da literatura sempre associaram a produção de quantidades desprezíveis de Hol à linhagem selvagem S. clavuligerus ATCC 27064. Por outro lado, cultivos em glicerol ou amido adicionados dos aminoácidos lisina, ácido alfa-aminoadípico, arginina e/ou ornitina, ou da diamina 1,3 diaminopropano, promovem produções significativas de compostos beta-lactâmicos, como CefC ou AC, com destaque para este último (DOMINGUES et al, 2010;LEITE et al, 2013). No presente trabalho, a produção de biocompostos por S. clavuligerus foi investigada em dois meios quimicamente definidos contendo diferentes fontes de carbono e nitrogênio.…”