Soybean-milk-coagulating activity of Bacillus pumilus derives from a serine proteinase.
A proteolytic enzyme from Bacillus pumilus strain TYO-67, which was able to coagulate the protein in soybean milk, was characterized enzymologically. The optimum pH and temperature for its activities were 9.0 and 50 degrees C, respectively. The enzyme was strongly believed to be a serine proteinase because it was completely inhibited by the addition of diisopropyl fluorophosphate or phenylmethanesulfonyl fluoride. Hammerstein milk casein, cytochrome c and soybean protein were good substrates for the enzyme. Seven cleavages were detected using the oxidized insulin B-chain as peptide substrate for the proteolytic specificity test of the serine proteinase from B. pumilus. The bonds most susceptible to the action of the serine proteinase from B. pumilus were Leu-15-Tyr-16. The mode of action on soybean milk protein by the enzyme from B. pumilus was also investigated. The acidic subunit in glycinin and the alpha-, alpha- and beta-subunits in beta-conglycinin were degraded during the enzyme reaction. However, the basic subunit in glycinin could not be degraded by the enzyme. The formation of coagula in soybean milk caused by the serine proteinase from B. pumilus was mainly due to the hydrophobic interaction.
Aoyama
M
M
Department of Bioscience and Biotechnology, Faculty of Agriculture, University of the Ryukyus, Okinawa, Japan.
Yasuda
M
M
Nakachi
K
K
Kobamoto
N
N
Oku
H
H
Kato
F
F
D001407:Bacillus / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D030262:Soybean Proteins / Q000378:metabolism*; D013025:Glycine max / Q000737:chemistry; D013379:Substrate Specificity; D020860:Subtilisin / Q000378:metabolism; D013696:Temperature
D007328:Insulin; D015842:Serine Proteinase Inhibitors; D030262:Soybean Proteins; D010450:Endopeptidases; D012697:Serine Endopeptidases; D020860:Subtilisin; C119936:soybean-milk-coagulating enzyme