Primary recovery of a bacteriocin-like inhibitory substance derived from Pediococcus acidilactici Kp10 by an aqueous two-phase system

Abstract

A polymer-salt aqueous two-phase system (ATPS) consisting of polyethylene-glycol (PEG) with sodium citrate was developed for direct recovery of a bacteriocin-like inhibitory substance (BLIS) from a culture of Pediococcus acidilactici Kp10. The influences of phase composition, tie-line length (TLL), volume ratio (VR), crude sample loading, pH and sodium chloride (NaCl) on the partition behaviour of BLIS was investigated. Under optimum conditions of ATPS, the purification of BLIS was achieved at 26.5% PEG (8000)/11% sodium citrate with a TLL of 46.38% (w/w), VR of 1.8, and 1.8% crude load at pH 7 without the presence of NaCl. BLIS from P. acidilactici Kp10 was successfully purified by the ATPS up to 8.43-fold with a yield of 81.18%. Given that the operation of ATPS is simple, environmentally friendly and cost-effective, as it requires only salts and PEG, it may have potential for industrial applications in the recovery of BLIS from fermentation broth.

Purification of Antilisterial Peptide (Subtilosin A) from Novel Bacillus tequilensis FR9 and Demonstrate Their Pathogen Invasion Protection Ability Using Human Carcinoma Cell Line

“…To demonstrate the antimicrobial activity, 10 μL test pathogens (10 7 –10 9 CFU/mL) were spread onto agar plates, 100 μL pH neutralized CFCS (pH 6.5) was added to each well and incubated (Supplementary Table S1). Antimicrobial activity was expressed as Arbitrary unit (AU) using following formula (Abbasiliasi et al, 2014). …”

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

“…Although a few studies using bacteriocin partitioning/recovery using PEG/salt-based ATPSs [26][27][28][29][30], and with an aqueous two-phase micellar system (ATPMS) [31,32], have been evaluated, PEG/NaPA systems composed of inorganic salts or choline-based salts have not been proposed before. In this context, and based on the advantages mentioned above, the present study investigated a PEG/NaPA based ATPS with Na 2 SO 4 or choline chloride ([Ch]Cl) as additives to extract bacteriocin from the fermented broth of Lactobacillus plantarum ST16Pa.…”

Purification of a Bacteriocin‐Like Inhibitory Substance Derived from Pediococcus acidilactici Kp10 by an Aqueous Micellar Two‐Phase System

“…3 Unfortunately, the conventional methods were laborious as they consisted of a multistep operation, expensive, time-consuming, low yield, and not efficient for industrial-scale requirement. 7 Therefore, there has been an increasing interest in seeking for alternative methods for bacteriocin purification such as through the liquid-liquid extraction method, which include the aqueous two-phase system (ATPS) [11][12][13] and aqueous micellar two-phase system (AMTPS) 14,15 These systems have been widely used for many target proteins extractions from unclarified microbial cultures, owing to their advantages: easy for scale-up, mildness for biological material, and controllable performance for process optimization-by varying the condition of the solution used. 16 In AMTPS, a variant of ATPS, a homogeneous aqueous micellar solution that composed of certain nonionic surfactants (with certain concentrations) becomes cloudy and separates into two macroscopic phases (micelle-rich phase and micellepoor phase) at certain temperatures (cloud point).…”