A new method for isolation of native α-lactalbumin from sweet whey

Pilot-scale purification of α-lactalbumin from enriched whey protein concentrate by anion-exchange chromatography and ultrafiltration

“…, which is exactly as expected for unmodified α-La, and the denaturation temperature T m was 66.1°C, which corresponds very well with the results obtained by others for the holo form of α-La (Konrad and Kleinschmidt 2008;Wang et al 2006;Tolkach et al 2005). Moreover, after converting calcium and α-La contents into moles, the molar ratio of Ca to α-La was 0.94, which is very close to 1.…”

“…Although this purity was slightly lower than that obtained at lab scale (98.6%), it is still the highest purity found when comparing with other pilot-scale purifications using WPC or WPI as raw material (Table 1). In addition, the total amount obtained (1100 g) is the highest amount obtained with reasonable purity (Table 1; Toro-Sierra et al 2013;Konrad and Kleinschmidt 2008;Kristiansen et al 1998). With the optimized method applied here, the total buffer volume used for recovery of α-La was 550 L (calculations not shown).…”

“…Schmidt and Poll (1991) reported that native α-La was highly resistant to trypsin while β-Lg was not. At lab scale, tryptic hydrolysis of β-Lg and discarding the fractions by membrane filtration seem suitable for achieving 100% purity of α-La (Lisak et al 2013;Cheison et al 2011); however, when upscaling, the purity drops to 90-95% and the recovery is quite low (Konrad and Kleinschmidt 2008). In order to achieve high purity, both at laboratory and pilot scale, chromatographic methods seem to be the best choice (Gurgel et al 2000;Ye et al 2000;Kristiansen et al 1998), and most often ion exchange chromatography is used.…”

Adsorptive behavior of α-lactalbumin on cation-exchange supermacroporous monolithic column

“…Several works have been conducted with the purpose of recovering and purifying whey proteins by different methods [8][9][10][11][12][13]. Ion-exchange chromatography (IEC) has been extensively employed in protein recovery and purification [14][15][16][17][18][19].…”

Influence of hydrolysis temperature and pH on the selective hydrolysis of whey proteins by trypsin and potential recovery of native alpha-lactalbumin

“…Schmidt and Poll (1991) showed that native a-La was highly resistant to tryptic digestion while native b-Lg was not. Indeed, Konrad and Kleinschmidt (2008), taking advantage of this selective susceptibility of b-Lg, used a combination of membrane technology and trypsin hydrolysis of the permeate to obtain a yield of a-La of 90e95% in purity. However, only 15% of the original a-La was recovered during hydrolysis at 42 C and pH 7.7.…”