Isolation and purification of bacterial proteinases by means of autofocusing

Sitkey, V.; Travěncová, Ivana; Minařík, Martin

doi:10.1016/0021-9673(91)85107-q

Cited by 3 publications

(2 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Many varieties of matrix-free, preparative, isoelectric focusing apparatuses have been developed. By using those apparatuses, enzymes, antibodies, peptides, and so on have been isolated without the addition of carrier ampholytes (12,(16)(17)(18)(19)(20)(21)(22)(23)(24). This approach has been referred to as autofocusing.…”

Section: Discussionmentioning

confidence: 99%

Development of a Large-Scale (50 L) Apparatus for Ampholyte-free Isoelectric Focusing (Autofocusing) of Peptides in Enzymatic Hydrolysates of Food Proteins

Hashimoto

Satô

Nakamura

et al. 2005

J. Agric. Food Chem.

View full text Add to dashboard Cite

It has been demonstrated that peptides in enzymatic hydrolysates of proteins can be fractionated on the basis of the amphoteric nature of the sample peptides, by a laboratory-scale isoelectric focusing apparatus, without adding a chemically synthesized carrier ampholyte. This approach is referred to as autofocusing. In the present study, a large-scale (up to 50 L) autofocusing apparatus was developed and tested. A tank (125 cm x 25 cm x 20 cm) was divided into 12 compartments by 11 plates, each with a window covered in a thin agarose gel layer supported by a nylon screen (100 mesh). The compartments at both ends were filled with 0.1 N phosphoric acid (anode) and 0.1 N NaOH (cathode), respectively, functioning as electrode compartments. The remaining compartments were used for sample compartments. Autofocusing was carried out at constant voltage according to two different methods. In method 1, all sample compartments were filled with a 1% water solution of casein or milk whey protein hydrolysates. In method 2, two compartments located in the center of the tank were filled with 5% sample solution and the others were filled with deionized water. Compositional and sequence analyses of the autofocusing fractions revealed that peptides in the two hydrolysates can be fractionated within 24 h by the present apparatus. Better fractionation was obtained by method 2, whereas enrichment of some peptides occurred by using method 1.

show abstract

Section: Discussionmentioning

confidence: 99%

Development of a Large-Scale (50 L) Apparatus for Ampholyte-free Isoelectric Focusing (Autofocusing) of Peptides in Enzymatic Hydrolysates of Food Proteins

Hashimoto

Satô

Nakamura

et al. 2005

J. Agric. Food Chem.

View full text Add to dashboard Cite

show abstract

“…By using this technique, peptide samples up to 50 L could be fractionated in the previous batch type apparatus (16). As the fractionation of peptides occurs in the agarose layer, the focusing time by using this system is considerably shorter than that by using large-scale free zone electrophoresis (17)(18)(19)(20)(21)(22)(23)(24). For industrial applications of this approach, much larger amounts of sample should be processed.…”

Section: Introductionmentioning

confidence: 99%

Development of Continuous Type Apparatus for Ampholyte-Free Isoelectric Focusing (Autofocusing) of Peptides in Protein Hydrolysates

Hashimoto

Satô

Nakamura

et al. 2006

J. Agric. Food Chem.

View full text Add to dashboard Cite

An apparatus for continuous fractionation of peptides on the basis of amphoteric nature of sample peptides was developed. A tank (66.5 cm x 8 cm x 8 cm, L x W x H) was divided into 12 compartments by a thin agarose gel layer. A drain tube (5.5 cm in length and 0.7 cm in i.d.) was fixed through the bottom of each compartment to give a height of 4 cm from the bottom. The tank with 12 compartments and electrodes was referred to as an autofocusing unit. The peptide solution or water was delivered to the sample compartments of the first unit. The solutions drained from the first unit were successively delivered to the second and third units. To the electrodes of three units, a direct electric current was applied. By using the present apparatus, peptides in casein hydrolysate can be continuously fractionated at least for 5 h. Better resolution was obtained in the second and third units.

show abstract

Isoelectric focusing in a multicompartment electrolyzer with zwitterionic membranes, exemplified by purification of glucoamylase

Wenisch¹,

Schneider²,

Hansen³

et al. 1993

Journal of Biochemical and Biophysical Methods

View full text Add to dashboard Cite

Isolation and purification of bacterial proteinases by means of autofocusing

Cited by 3 publications

References 4 publications

Development of a Large-Scale (50 L) Apparatus for Ampholyte-free Isoelectric Focusing (Autofocusing) of Peptides in Enzymatic Hydrolysates of Food Proteins

Development of a Large-Scale (50 L) Apparatus for Ampholyte-free Isoelectric Focusing (Autofocusing) of Peptides in Enzymatic Hydrolysates of Food Proteins

Development of Continuous Type Apparatus for Ampholyte-Free Isoelectric Focusing (Autofocusing) of Peptides in Protein Hydrolysates

Isoelectric focusing in a multicompartment electrolyzer with zwitterionic membranes, exemplified by purification of glucoamylase

Contact Info

Product

Resources

About