Purification of glucosyltransferase from cell-lysate of Streptococcus mutans by counter-current chromatography using aqueous polymer two-phase system

Amino acids partitioning in aqueous two-phase system of polypropylene glycol and magnesium sulfate

“…(m i γ i ) top = (m i γ i ) bot (11) The fitting parameters of the model were obtained as: A p-p = −0.785, A p-s = −0.465 and A s-s = 0.433 kg mol −1 .…”

“…The cross-axis coil planet centrifuge (CPC) allows satisfactory levels of stationary phase retention for the aqueous-aqueous polymer phase systems so that it can be effectively used for the preparative separation of proteins [8]. With some PEG-salt systems, this method has been used successfully for the separations of a variety of protein samples [9][10][11]. The CCC method using aqueous two-phase systems could also be applied for separation of the amino acids.…”

Purification of Proteins From Cell-Culture Medium or Cell-Lysate by High-Speed Counter-Current Chromatography Using Cross-Axis Coil Planet Centrifuge

“…This can be done spectrophotometrically by a simple test tube experiment as described elsewhere [26]. For the determination of partition coefficient of S. mutans cell-lysate (K SM ), the total UV at 220 nm of the proteins in both upper and lower phases were measured with a UV-1200 spectrophotometer (Shimadzu, Kyoto, Japan).…”

“…In the past the CCC separations of basic histones, serum proteins [25], and profilin-actin complex from crude Acanthamoeba extract [23] were performed using PEGdextran two-phase systems. We have demonstrated the purification of glucosyltransferase (GTF) from Streptococcus mutans cell-lysate and S. sobrinus culture medium [26,27] using a 7.7% PEG 8000-4.0% dextran T500 solvent system. After HSCCC purification, these polymers were removed from CCC fractions using a hydroxyapatite chromatography.…”

Aqueous–aqueous two-phase systems composed of low molecular weight of polyethylene glycols and dextrans for counter-current chromatographic purification of proteins

“…In the past the CCC separations of basic histones, serum proteins [20], and profilin-actin complex from crude Acanthamoeba extract [18] were performed using PEG-dextran two-phase systems. Recently, we have demonstrated the purification of glucosyltransferase (GTF) from S. mutans cell-lysate and S. sobrinus culture medium [21,22] using a 7.7% PEG 8000-4.0% dextran T500 solvent system. After CCC purification, these polymers were removed from CCC fractions by hydroxyapatite chromatography.…”