Bacteriocin partitioning from a clarified fermentation broth of Lactobacillus plantarum ST16Pa in aqueous two-phase systems with sodium sulfate and choline-based salts as additives

Sabo, Sabrina da Silva; Lopes, André Moreni; Santos-Ebinuma, Valéria de Carvalho; Rangel-Yagui, Carlota de Oliveira; Oliveira, Ricardo Pinheiro de Souza

doi:10.1016/j.procbio.2017.11.018

Cited by 21 publications

(14 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Some previous studies have shown that the negative charge of NaPA can interact with the oppositely charged sites of biomolecules through electrostatic interactions, destabilizing or changing their molecular structure and decreasing their biological activity [31,41]. However, at the concentrations studied, GFP maintained its fluorescence activity, which is consistent with previous findings obtained for clavulanic acid, where no significant losses of activity were observed after the exposure of the biomolecule to aqueous solutions of NaPA 8000 at 5 and 15 wt% [42].…”

Section: Gfp Fluorescence Stabilitysupporting

confidence: 90%

Effect of electrolytes as adjuvants in GFP and LPS partitioning on aqueous two-phase systems: 1. Polymer-polymer systems

Lopes

Molino

Santos-Ebinuma

et al. 2018

Separation and Purification Technology

Self Cite

View full text Add to dashboard Cite

The production of recombinant biopharmaceuticals is highly dependent of a proper choice of the downstream processing stages. Particularly, the purification that must ensure that all the endotoxins (lipopolysaccharides-LPS) are efficiently removed from the final product. The efficient removal of LPS has a direct impact on the manufacturing of therapeutic biopharmaceuticals, since LPS is naturally presented in Gram-negative bacterial expression systems. In order to provide a more simple and faster technique for the purification of green fluorescent protein (GFP) and LPS removal, aqueous two-phase systems (ATPS) composed of polyethylene glycol (PEG) and poly(acrylic acid) (NaPA) and electrolytes were studied. Firstly, the binodal curves of PEG/ NaPA + salt systems were established using NaCl, Li 2 SO 4 , KI, and KNO 3 as additives. It was demonstrated that the formation of ATPS is enhanced following the anion tendency: SO 4 2− > Cl − ≫ NO 3 − > I −. The stability of GFP in the presence of all different phase forming agents (polymers and salts) was evaluated, and the high biocompatibility of these ATPS demonstrated by the maintenance of GFP fluorescence in all conditions under study. Then, the GFP extraction and LPS removal aptitude of each ATPS was investigated. GFP and LPS were preferentially partitioned into the top (PEG-rich) phase (K GFP > 20), but with a removal of 35% of LPS was attained. Hydrophobic and electrostatic interactions were found to be the major driving forces for GFP partitioning and LPS removal. Moreover, a new effect was found, the presence of high loads of LPS can affect (decrease) the K GFP values (K GFP without LPS > K GFP with 10 4 EU/mL > K GFP with 10 6 EU/mL). The ATPS with best GFP extraction performance was selected for the recovery directly from the cell lysates of Escherichia coli. In this experiment, the system composed of 12 wt% PEG 1000 g/mol, 12 wt% NaPA 8000 g/mol, and 0.25 M Li 2 SO 4 lead to a LPS removal (REM LPS) of 13%, K GFP of 20.2, and high selectivity relatively to the total proteins (S = 20), since the majority of contaminants proteins were preferentially partitioned into the bottom (NaPArich) phase (purification factor of 4-fold). It is here demonstrated that the ATPS composed of PEG/NaPA + salts as additives can be used as a first step for the recovery of GFP and removal of contaminants (LPS in part, and most contaminant proteins) from cell lysates by applying a system with low polymer content and using mild conditions.

show abstract

Section: Gfp Fluorescence Stabilitysupporting

confidence: 90%

Effect of electrolytes as adjuvants in GFP and LPS partitioning on aqueous two-phase systems: 1. Polymer-polymer systems

Lopes

Molino

Santos-Ebinuma

et al. 2018

Separation and Purification Technology

Self Cite

View full text Add to dashboard Cite

show abstract

“…These results might have been due to removal of protein contaminants responsible for suppression of pediocin PA-1 antimicrobial activity or its inactivation. Likewise, Sabo et al (2018), using ATPS for bacteriocin partitioning from clarified broth of Lb. plantarum ST16Pa, demonstrated the potential of this system as an initial step for bacteriocin recovery and purification.…”

Section: Purification Of Bacteriocin St16 Obtained In Mrs Broth and Imentioning

confidence: 99%

Bacteriocin production by Lactobacillus plantarum ST16Pa in supplemented whey powder formulations

Sabo

Converti

Ichiwaki

et al. 2019

Journal of Dairy Science

View full text Add to dashboard Cite

Whey, the main by-product of the dairy industry, is frequently disposed of in the environment without any treatment due to the high cost of this process. Alternatively, whey can be used as a medium to culture lactic acid bacteria and produce value-added products such as bacteriocins. In this work, we attempted to improve bacteriocin production by Lactobacillus plantarum ST-16Pa in a whey powder formulation supplemented with additional sources of carbon, nitrogen, and vitamin B 12 at different levels and varying the agitation intensity according to a Plackett-Burman experimental design. Only the addition of tryptone positively influenced the production of this bacteriocin. The results allowed us to identify a supplemented whey formulation, comprising 150 g/L of whey total solids plus 10 g/L of tryptone and soybean extract, whose fermentation by Lb. plantarum ST16Pa in shake flasks under agitation at 150 rpm led to a cell-free supernatant with an antimicrobial activity against Listeria innocua 6a CLIST 2865 (inhibition zone of 13.23 mm) close to that previously obtained in de Man, Rogosa and Sharpe medium by other authors. These results are significant considering that the same strain cultured in cheese whey did not previously display any antimicrobial activity.

show abstract

“…Hence, the purification factor decreased with increasing the PEG molecular weight to 6000 and 8000 due to the excluded volume effect. Sabrina da Silva Sabo et al [12] reported that a low concentration of PEG could increase the purification factor of BLIS in an aqueous two phase system. Hence, 10% (w/w) of PEG 4000 with the highest purification factor was used in further experiments.…”

Section: Discussionmentioning

confidence: 99%

Recovery of a Bacteriocin-Like Inhibitory Substance from Lactobacillus bulgaricus FTDC 1211 Using Polyethylene-Glycol Impregnated Amberlite XAD-4 Resins System

Aziz

Abbasiliasi

et al. 2020

Molecules

View full text Add to dashboard Cite

Lactobacillus bulgaricus is a LAB strain which is capable of producing bacteriocin substances to inhibit Staphylococcus aureus. The aim of this study was to purify a bacteriocin-like inhibitory substance (BLIS) produced by L. bulgaricus FTDC 1211 using an aqueous impregnated resins system consisting of polyethylene-glycol (PEG) impregnated on Amberlite XAD4. Important parameters influencing on purification of BLIS, such as the molecular weight and concentration of PEG, the concentration and pH of sodium citrate and the concentration of sodium chloride, were optimized using a response surface methodology. Under optimum conditions of 11% (w/w) of PEG 4000 impregnated Amberlite XAD4 resins and 2% (w/w) of sodium citrate at pH 6, the maximum purification factor (3.26) and recovery yield (82.69% ± 0.06) were obtained. These results demonstrate that AIRS could be used as an alternate purification system in the primary recovery step.

show abstract

Bacteriocin partitioning from a clarified fermentation broth of Lactobacillus plantarum ST16Pa in aqueous two-phase systems with sodium sulfate and choline-based salts as additives

Cited by 21 publications

References 44 publications

Effect of electrolytes as adjuvants in GFP and LPS partitioning on aqueous two-phase systems: 1. Polymer-polymer systems

Effect of electrolytes as adjuvants in GFP and LPS partitioning on aqueous two-phase systems: 1. Polymer-polymer systems

Bacteriocin production by Lactobacillus plantarum ST16Pa in supplemented whey powder formulations

Recovery of a Bacteriocin-Like Inhibitory Substance from Lactobacillus bulgaricus FTDC 1211 Using Polyethylene-Glycol Impregnated Amberlite XAD-4 Resins System

Contact Info

Product

Resources

About