Methods in Molecular Biology • 16 Enzymes of Molecular Biology

(Nancy Kaufman) #1
CHAPTER 14

Pronase (EC 3.4.24.4)


Patricia J. Sweeney and John M. Walker



  1. Introduction
    Pronase is the name given to a group of proteolytic enzymes that are
    produced in the culture supematant of Streptomyces griseus K- 1 (1-3).
    Pronase is known to contain at least ten proteolytic components: five
    serine-type proteases, two Zn 2+ endopeptidases, two Zn2+-leucine ami-
    nopeptidases, and one Zn 2+ carboxypeptidase (4,5). Pronase therefore
    has very broad specificity, and is used in cases where extensive or
    complete degradation of protein is required. It has been used, for
    example, to reveal the protein components of cell organelles by the
    hydrolysis of tissue slices (6), and as an alternative to proteinase K to
    remove protein during plasmid DNA (7), chromosomal DNA (8), and
    RNA isolation (9-11). Another use of pronase is the production of a
    protein hydrolysate suitable for amino acid analysis (12,13).
    Traditionally, protein hydrolysates for amino acid analysis are pro-
    duced by hydrolysis in 6N HC1. However, this method has the disad-
    vantage that tryptophan is totally destroyed, serine and threonine
    partially (5-10%) destroyed, and most importantly, asparagine and
    glutamine are hydrolyzed to the corresponding acids. Digestion of the
    protein/peptide with pronase overcomes these problems, and is par-
    ticularly useful when the concentration of asparagine and glutamine is
    required. The use of pronase for this application is detailed in Section

  2. The use of pronase in the preparation of optically active amino acids
    has been described. The hydrolysis of amino amides by pronase results
    in high-rate significant enantioselectivity and a high degree of conver-


From: Methods in Molecular Biology, VoL 16: Enzymes of Molecular Biology
Edited by: M. M. Burrell Copyright ©1993 Humana Press Inc., Totowa, NJ

271
Free download pdf