Abstract
The production of a bacteriocin-like substance with antimicrobial activity, named peocin, by the probiotic Paenibacillus ehimensis NPUST1 was previously reported by our laboratory. The present study aimed to identify peocin and increase the peocin yield by heterologous expression in Escherichia coli BL21(DE3). Peocin was identified as a DNA starvation/stationary phase protection protein, also called DNA-binding protein from starved cells (Dps), by gel overlay and LC-MS/MS analysis. For mass production of peocin, fed-batch cultivation of E. coli was performed using a pH-stat control system. Purification by simple nickel affinity chromatography and dialysis yielded 45.3 mg of purified peocin from a 20-mL fed-batch culture (49.3% recovery). The biological activity of the purified peocin was confirmed by determination of the MIC and MBC against diverse pathogens. Purified peocin exhibited antimicrobial activity against aquatic, food spoilage, clinical and antibiotic-resistant pathogens. In an in vivo challenge test, zebrafish treated with purified peocin exhibited significantly increased survival rates after A. hydrophila challenge. The present study is the first to show the antimicrobial activity of Dps and provides an efficient strategy for production of bioactive peocin, which will aid the development of peocin as a novel antimicrobial agent with potential applications in diverse industries.
Highlights
Since the first antibiotic penicillin was accidentally discovered by Alexander Fleming in 1928, investigations on the isolation of new antibiotics or chemical modification of antibiotics have rapidly developed over the past few decades
These results suggest that the bacteriocin-like substance peocin from P. ehimensis NPUST1 is a DNA starvation/stationary phase protection protein
The results showed a distinct inhibition zone, convincingly demonstrating that peocin is a bacteriocin-like substance produced by P. ehimensis NPUST1 and that the recombinant peocin possesses antimicrobial activity (Figure 3C)
Summary
Since the first antibiotic penicillin was accidentally discovered by Alexander Fleming in 1928, investigations on the isolation of new antibiotics or chemical modification of antibiotics have rapidly developed over the past few decades. The indiscriminate use of antibiotics has led to problems, including the rapid spread of multidrug-resistant (MDR) or pandrug-resistant (PDR) pathogens, reduced effectiveness of antibiotic treatment, damage to microbial ecosystems in the environment, and the emergence of antibiotic-resistant organisms from the consumption of antibiotic residues in food [1,2,3]. Lactobacillus plantarum with pediocin-like activity isolated from cattle shows potential probiotic traits of bile and low-pH tolerance, suggesting that this species could be used to overcome the problem of antibiotic resistance in veterinary medicine [11] Bacteriocins such as nisin, pediocin A, and divercin were used to prevent necrotic enteritis in poultry caused by Clostridium perfringens [12]. Nisin-producing Lactococcus lactis strain A5 isolated from broadhead catfish (Clarias macrocephalus) exhibited antagonistic activity against pathogens, suggesting the potential applications of this strain as a feed component in aquaculture [13]
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