Cloning, Production, and Functional Expression of the Bacteriocin Enterocin A, Produced by Enterococcus faecium T136, by the Yeasts Pichia pastoris, Kluyveromyces lactis, Hansenula polymorpha, and Arxula adeninivorans

Borrero, Juan; Kunze, Gotthard; Jiménez, Juan J.; Böer, Erik; Gútiez, Loreto; Herranz, Carmen; Cintas, Luis M.; Hernández, Pablo E.

doi:10.1128/aem.00530-12

Cited by 34 publications

(20 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several studies have been performed on heterologous expression to improve the yield of bacteriocins and other antimicrobial compounds [18,19]. In this study, it has been clearly demonstrated that the production of recombinant plantaricin can be achieved from a heterologous host, E. coli.…”

Section: Discussionmentioning

confidence: 96%

Scaling Up the Production of Recombinant Antimicrobial Plantaricin E from a Heterologous Host, Escherichia coli

Pal

Srivastava

2015

Probiotics & Antimicro. Prot.

View full text Add to dashboard Cite

Enhanced production of heterologously expressed plantaricin (plnE) from Escherichia coli BL21 (DE3) was achieved from a small- to large-scale batch culture. Starting from a 15-ml shake-flask culture grown in Luria-Bertani (LB) broth, the protein expression could be scaled up using 50 ml, 100 ml, 1 l, and 2 l batch culture. Using similar condition, plantaricin E (PlnE) was successfully expressed in a 30-l stirred fermenter. The protein was expressed as TRX-(His)6-fusion protein and separated by Ni(2+) affinity chromatography. Growth in two complex media, LB and Terrific broth (TB), was optimized and compared for the production of PlnE, which was higher in LB in comparison with that of TB. In the fermenter, 140 and 180 mg of PlnE could be produced from 12 l of culture volume at 30 and 25 °C, respectively. The yield of heterologously purified PlnE was found to be 1.2-1.5%, which was much higher in comparison with the plantaricins produced from the native strain of Lactobacillus plantarum (0.3-0.7%). Overproduction of PlnE with the help of heterologous expression can overcome the constraint of the low yield from producer strain and provides an easy and low-cost strategy for large-scale production.

show abstract

Section: Discussionmentioning

confidence: 96%

Scaling Up the Production of Recombinant Antimicrobial Plantaricin E from a Heterologous Host, Escherichia coli

Pal

Srivastava

2015

Probiotics & Antimicro. Prot.

View full text Add to dashboard Cite

show abstract

“…In addition, this bacteriocin has been expressed in Lc. lactis (Martín et al 2007) and in yeasts (Borrero et al 2012) to improve food preservation. In this study, the MIC of enterocin A is lower than other known Fig.…”

Section: Discussionmentioning

confidence: 99%

Enhanced bactericidal effect of enterocin A in combination with thyme essential oils against L. monocytogenes and E. coli O157:H7

Ghrairi

Hani

2013

J Food Sci Technol

View full text Add to dashboard Cite

The combined effects of enterocin A with Thymus vulgaris essential oils (EOs) against Listeria monocytogenes and Escherichia coli O157:H7 were investigated in vitro by enumeration of surviving populations of testing pathogens and minimal inhibitory concentration (MIC) determination. Enterocin A was purified to homogeneity by RP-HPLC from the culture fluid of Enterococcus strain and thyme EOs were extracted from local Thymus vulgaris plants. The major constituent of thyme EOs oils determined by GC-MS was thymol (78.4 %). Combination of enterocin A with thyme EOs showed an enhanced bactericidal effect against Listeria monocytogenes. Checkerboard assay and isobologram construction displayed a synergistic interaction between these compounds against Listeria (FIC index <0.5). Moreover, the MIC value of enterocin A has fallen fivefold (from 4.57 to 0.9 μg/ml), while the MIC of thyme EOs decreased threefold (from 3.6 to 1.2 μg/ml). Treatments with enterocin A alone did not affect the growth of the enteric pathogen E. coli O157:H7. However, the addition of thyme EOs and enterocin A yielded a synergistic antimicrobial effect against E. coli (MIC thyme EOs decrease from 2.2 to 0.71 μg/ml). This is the first report on the combined effect of enterocin A and thyme EOs against food pathogen bacteria. This combination could be useful in food bio-preservation.

show abstract

“…However, verifying the antimicrobial activity of putative bacteriocins can be difficult due to the potentially narrow activity spectra and highly diverse mechanisms 4,37 . Additionally, traditional methods of natural bacteriocin isolation as well as heterologous expression strategies are complicated by purification limitations and low yield 38–40 . Here we describe a complete strategy by which de novo mining of bacteriocins can be parsed using a biophysical algorithm to identify minimally active bacteriocin peptide candidates.…”

Section: Discussionmentioning

confidence: 99%

Novel antimicrobial peptide discovery using machine learning and biophysical selection of minimal bacteriocin domains

Fields

Freed

Carothers

et al. 2018

Preprint

View full text Add to dashboard Cite

17Bacteriocins are ribosomally produced antimicrobial peptides that represent an untapped 18 source of promising antibiotic alternatives. However, inherent challenges in isolation and 19 identification of natural bacteriocins in substantial yield have limited their potential use as 20 viable antimicrobial compounds. In this study, we have developed an overall pipeline for 21 bacteriocin-derived compound design and testing that combines sequence-free prediction of 22 bacteriocins using a machine-learning algorithm and a simple biophysical trait filter to generate 23 minimal 20 amino acid peptide candidates that can be readily synthesized and evaluated for 24 activity. We generated 28,895 total 20-mer peptides and scored them for charge, a-helicity, and 25 hydrophobic moment, allowing us to identify putative peptide sequences with the highest 26 potential for interaction and activity against bacterial membranes. Of those, we selected sixteen 27 sequences for synthesis and further study, and evaluated their antimicrobial, cytotoxicity, and 28 hemolytic activities. We show that bacteriocin-based peptides with the overall highest scores for 29 our biophysical parameters exhibited significant antimicrobial activity against E. coli and P. 30 aeruginosa. Our combined method incorporates machine learning and biophysical-based 31 minimal region determination, to create an original approach to rapidly discover novel 32 bacteriocin candidates amenable to rapid synthesis and evaluation for therapeutic use.

show abstract

Cloning, Production, and Functional Expression of the Bacteriocin Enterocin A, Produced by Enterococcus faecium T136, by the Yeasts Pichia pastoris, Kluyveromyces lactis, Hansenula polymorpha, and Arxula adeninivorans

Cited by 34 publications

References 44 publications

Scaling Up the Production of Recombinant Antimicrobial Plantaricin E from a Heterologous Host, Escherichia coli

Scaling Up the Production of Recombinant Antimicrobial Plantaricin E from a Heterologous Host, Escherichia coli

Enhanced bactericidal effect of enterocin A in combination with thyme essential oils against L. monocytogenes and E. coli O157:H7

Novel antimicrobial peptide discovery using machine learning and biophysical selection of minimal bacteriocin domains

Contact Info

Product

Resources

About