title: Scaling Up the Production of Recombinant Antimicrobial Plantaricin E from a Heterologous Host, Escherichia coli.
abstract: 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.
authors.affiliation: Department of Genetics, University of Delhi, South Campus, New Delhi, 110021, India.
authors.forename: Gargi
authors.initials: G
authors.lastname: Pal
authors.forename: Sheela
authors.initials: S
authors.lastname: Srivastava
mesh_terms: D000890:Anti-Infective Agents / Q000737:chemistry*; D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics; D061249:Batch Cell Culture Techniques; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial; D048191:Lactiplantibacillus plantarum / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D059467:Transcriptome
chemical_list: D000890:Anti-Infective Agents; D001430:Bacteriocins; D011994:Recombinant Proteins