Inducing Effect of Diamines on Transcription of the Cephamycin C Genes from the lat and pcbAB Promoters in Nocardia lactamdurans

Abstract

The diamines putrescine, cadaverine, and diaminopropane stimulate cephamycin biosynthesis in Nocardia lactamdurans, in shake flasks and fermentors, without altering cell growth. Intracellular levels of the P7 protein (a component of the methoxylation system involved in cephamycin biosynthesis) were increased by diaminopropane, as shown by immunoblotting studies. Lysine-6-aminotransferase and piperideine-6-carboxylate dehydrogenase activities involved in biosynthesis of the α-aminoadipic acid precursor were also greatly stimulated. The diamine stimulatory effect is exerted at the transcriptional level, as shown by low-resolution S1 protection studies. The transcript corresponding to the pcbAB gene and to a lesser extent also the lat transcript were significantly increased in diaminopropane-supplemented cultures, whereas transcription from the cefD promoter was not affected. Coupling of the lat and pcbABpromoters to the reporter xylE gene showed that expression from the lat and pcbAB promoters was increased by addition of diaminopropane in Streptomyces lividans. Intracellular accumulation of diamines in Nocardia may be a signal to trigger antibiotic production.

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

“…In contrast, the glycerol feeding strongly enhanced CA as previously reported [21]. Interestingly, the variable concentration of 1,3-diaminopropane in the culture changes that connected production in the favor of either compound [13,22].…”

“…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].…”

Dissociation of cephamycin C and clavulanic acid biosynthesis by 1,3-diaminopropane inStreptomyces clavuligerus

“…Several researchers have promoted production of betalactam antibiotics by adding 1,3-diaminopropane to the culture medium (Leitão et al 1999;Martín et al 2011;Leite, Cavallieri and Araujo 2013). Leitão et al (1999) and Leite, Cavallieri and Araujo (2013) reported a 100% increase in CephC production by S. clavuligerus by adding 5 g l −1 1,3-diaminopropane to the culture medium. Martín et al (2011) added low quantities of 1,3diaminopropane (0.35 and 0.75 g l −1 ) to a Penicillium chrysogenum culture medium and observed, too, that penicillin production increased by approximately 100%.…”

“…Polyamines also stabilize macromolecules, proteins and membranes modulating electrostatic proteinprotein interactions and affecting DNA-protein interactions (Bassard et al 2010). Leitão et al (1999) studied the effect of 1,3diaminopropane on Amycolatopsis lactamdurans and found that this diamine induces gene expression and stabilizes messenger RNAs. These authors reported, also, high levels of LAT and 1piperideina-6-carboxylate dehydrogenase (P6C dehydrogenase) enzymes and P7 protein.…”

“…The use of glycerol and vegetable oils as carbon source or ornithine as nitrogen source increases CA production (Domingues et al 2010;Teodoro et al 2010;Bellão et al 2013;Salem-Bekhit, Alanazi and Alsarra 2013). Also, additions of diamines, L-lysine or alpha-aminoadipic acid promote the production of beta-lactam antibiotics in actinomycetes (Fang, Keables and Demain 1996;Leitão et al 1999;Demain and Vaishnav 2006;Leite, Cavallieri and Araujo 2013). On the other hand, ammonium ions and inorganic phosphate can inhibit CephC and CA biosynthesis (Bibb 2005;Demain and Vaishnav 2006).…”