Production and Purification of Acid Protease from the Thermophilic Fungus,
            <i>Penicillium duponti</i>
            K1014

Hashimoto, Hikotaka; Kaneko, Yukio; Iwaasa, Takashi; Yokotsuka, Tamotsu

doi:10.1128/am.25.4.584-588.1973

Cited by 10 publications

(1 citation statement)

References 13 publications

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“…The crude protease preparation of Penicillium duponti K 1014 was provided by Kikkoman Shoyu Company, Noda, Japan, and consisted of the crude precipitate obtained by adding three volumes of 95% ethanol to one volume of clarified fermented broth, followed by centrifugation and drying in vacuo (Hashimoto et al, 1973b). The dried crude enzyme was a brownish powder, having a specific activity of 618 units/mg of protein.…”

Section: Methodsmentioning

confidence: 99%

Purification and properties of the thermostable acid protease of Penicillium duponti

Emi¹,

Myers²,

Iacobucci³

1976

Biochemistry

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An acid protease produced by the thermophilic fungus Penicillium duponti K 1014 has been purified by consecutive ion-exchange and gel permeation chromatography, and crystallized from aqueous acetone solution. The purified endopeptidase gave a symmetrical schlieren peak by sedimentation velocity, and was found to be homogeneous upon disc gel electrophoresis at pH 9.5. The enzyme was most active at pH 2.5 against milk casein and showed high thermostability. An isoelectric point of 3.81 was found by isoelectric focusing. A minimum molecular weight of 41 590 was calculated from the amino acid composition, adopting an arginine content of one residue per mole of enzyme. This minimum molecular weight is in good agreement with the value of 41 000 previously found by gel permeation (Hashimoto, H., Iwaasa, T., and Yokotsuka, T. (1973), Appl. Microbiol. 25, 578). Besides the thermostability, the purified P. duponti protease differs from other well-characterized acid proteases in that it contains carbohydrate, 4.33% expressed as glucose. The enzyme was not affected by p-bromophenacyl bromide, but was completely inactivated by alpha-diazo-p-bromoacetophenone, diazoacetyl-DL-norleucine methyl ester, and diazoacetylglycine ethyl ester, in the presence of Cu2+. The complete inactivation of the protease by diazoacetyl-DL-norleucine methyl ester resulted in the specific incorporation of 1 mol of norleucine/mol of enzyme. On the basis of similar behavior of other acid proteases toward this inactivator, the results suggest the presence at the active site of an unusually reactive carboxyl group, involved in the catalytic function. The naturally occurring pepsin inhibitor of Streptomyces naniwaensis [Murao, S., and Satoi, S. (1970), Agric. Biol. Chem. 34, 1265] inhibited also the protease, at a threefold molar excess with respect to the enzyme.

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Section: Methodsmentioning

confidence: 99%

Purification and properties of the thermostable acid protease of Penicillium duponti

Emi¹,

Myers²,

Iacobucci³

1976

Biochemistry

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Proteolysis by toxigenic Aspergillus nidulans from Nigerian palm produce

1987

Nahrung

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The submerged cultures of Aspergillus nidulans had optimal growth and protease production at 37 degrees C and within 6 days of incubation. A rapid drop in pH of the growth medium from 6.9 to 4.8 and a subsequent gradual rise was recorded with the period of incubation. The acid-protease produced was purified by a combination of ethanolic precipitation, ultrafiltration and fractionation on DEAE-cellulose and Sephadex G-200. A single peak showing protease activity was subsequently obtained with a 16-fold increase in specific activity and a recovery value of 36%. The purified enzyme had optimal activity on casein and gelatin at pH 5.4 and a temperature of 40 degrees C.

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Gewinnung und Charakteristik von Proteasen aus Thermoactinomyces vulgaris I. Proteasen aus thermophilen Mikroorganismen (Literaturüberblick)

Ruttloff

Klingenberg

Behnké

et al. 1978

J of Basic Microbiology

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Es wird ein Literaturüberblick zur Gewinnung und Charakteristik von Proteasen aus thermophilen Mikroorganismen gegeben. Nach einem allgemeinen Überblick zur Problematik der Thermophilie wird auf die von Schimmelpilzen und Bakterien produzierten proteolytischen Enzyme eingegangen. Besonders hervorgehoben werden die Protease aus Streptomyces rectus var. proteolyticus, die „Aminopeptidase I”︁ aus Bacillus stearothermophilus sowie das Thermolysin aus B. thermoproteolyticus.

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Production and Purification of Acid Protease from the Thermophilic Fungus, Penicillium duponti K1014

Cited by 10 publications

References 13 publications

Purification and properties of the thermostable acid protease of Penicillium duponti

Purification and properties of the thermostable acid protease of Penicillium duponti

Proteolysis by toxigenic Aspergillus nidulans from Nigerian palm produce

Gewinnung und Charakteristik von Proteasen aus Thermoactinomyces vulgaris I. Proteasen aus thermophilen Mikroorganismen (Literaturüberblick)

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