Proteomic insight into the primycin fermentation process of Saccharomonospora azurea

Abstract

Saccharomonospora azurea SZMC 14600 is a member of the family Pseudonocardiaceae exclusively used for industrial scale production of primycin a large 36-membered non-polyene macrolide lactone antibiotic belonging to the polyketide class of natural products. Even though maximum antibiotic yield has been achieved by empirically optimized two-step fermentation process, little is known about the molecular components and mechanisms underlying the efficient antibiotic production. In order to identify differentially expressed proteins (DEPs) between the pre-and main-fermentation stages of primycin, comparative 2D-PAGE experiments were performed. In total, 98 DEP spots were reproducibly detected, out of which four spots were excised from gels, and identified through MALDI-TOF/TOF mass spectrometry. Peptide mass fingerprint analysis revealed peptide matches to HicB antitoxin for the HicAB toxinantitoxin system (EHK86651), to a nucleoside diphosphate kinase regulator ((Ndk; EHK81899) and two other proteins with unknown function (EHK88946 and EHK86777).

Antimicrobial Biosynthetic Potential and Phylogenetic Analysis of Culturable Bacteria Associated with the Sponge Ophlitaspongia sp. from the Yellow Sea, China

“…HMA015 had both PKS and NRPS genes. The PKS gene sequence from HMA005 exhibited the closest amino acid similarity (99.11%) with sequences from Saccharomonospora azurea , which produces primycin [ 77 ]. The NRPS gene sequences of HMA035 and HMA027 exhibited 96.2% and 97.5% similarities at the amino acid level, respectively, with that of Streptomyces sp.…”

Structural and functional comparison of Saccharomonospora azurea strains in terms of primycin producing ability

“…Bacterial cultures were stored as a suspension in Luria Bertani (LB) broth with 20% (v/v) glycerol at − 80 °C. Culture conditions were carried out according to Valasek et al ( 2016 ). Briefly 1 mL bacterial cell suspension was inoculated into 50 mL seed medium containing 3% (w/v) soy flour, 4.2% (w/v) water soluble starch, 0.36% (w/v) NaCl, 0.6% (w/v) CaCO 3 , 0.5% (w/v) sunflower oil, pH 8.0, and incubated for 2 days at 37 °C in an orbital shaker at 200 rpm.…”

Understanding the Role of Fatty Acid Substrates on Primycin Biosynthesis by Saccharomonospora azurea During Batch Fermentation

“…Culture conditions were carried out according to Valasek et al with modifications. 30 Briefly frozen stock cultures were thawed at room temperature then inoculated directly into sterile 300 mL Erlenmeyer flask, containing 50 mL Luria-Bertani broth and incubated for 2 days at 37°C in an orbital shaker at 200 rpm. After 2 days 1 mL bacterial cell suspension was inoculated into sterile 300 mL Erlenmeyer flask, containing 35 mL primycin fermentation medium consisting of 4% soy flour, 4% water soluble starch, 0.3% NaCl, 0.5% CaCO 3 , 0.6% sunflower oil, 0.1% KH 2 PO 4 and 0.3% to 0.6% selected FA according to the experiment (stearic acid [C18:0], palmitic acid [C16:0], lauric acid [C12:0], capric acid [C10:0], enanthic acid [C7:0], caproic acid [C6:0], or butyric acid [C4:0]).…”

“…In this present study various FA compositions of the primycin fermentation media were investigated in order to determine their effect on antibiotic production. As a first step of our FA experiments the original 3 g/L stearic acid-containing fermentation media 30 was compared with FA-free media and with 3 g/L palmitic acid-containing fermentation media in order to determine if a slight modification in FA composition-changing the 18 carbon length to 16 carbon-in the growth medium has any beneficial effect on primycin production. During fermentation primycin concentrations were measured from the fifth day till the seventh day by the high-performance liquid chromatography-diode array detector-mass spectrometry detection (HPLC-DAD-MSD) method ( Figure 2).…”