Studies on Bacterial Protease

TSURU, Daisuke; Kira, Heizo; Yamamoto, Takehiko; Fukumoto, Juichiro

doi:10.1271/bbb1961.30.856

Cited by 22 publications

(2 citation statements)

References 3 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The bacterial neutral proteases are zinc metalloenzymes (McConn et al, 1964;Tsuru et al, 1966;Hiramatsu, 1967;Matheson and Armstrong;Keay, 1969;Griffin and Prescott, 1970;Keay et al, 1971;Naicajima et al, 1974). Thermolysin, B. subtilis neutral protease, and the neutral protease of Aeromonas proteolytica all actively hydrolyze esters, a feature hitherto unrecognized.…”

Section: Discussionmentioning

confidence: 99%

Esterase activity of zinc neutral proteases

Holmquist¹,

Vallée²

1976

Biochemistry

View full text Add to dashboard Cite

The hydrolysis of a series of depsipeptides demonstrates that the zinc neutral endopeptidases of bacteria are active esterases. Esters such as BzGly-OPhe-Ala, BzGly-OLeu-Ala, and FA-Gly-OLeu-NH2 are hydrolyzed at rates three- to eightfold slower than are their exact peptide analogues, when hydrolyzed by thermolysin, Bacillus subtilis neutral protease and the neutral protease from Aeromonas proteolytica. Ester hydrolysis by zinc neutral proteases follows the characteristic preference for hydrophobic amino acids adjacent to the site of cleavage, discerned from the hydrolysis of peptide substrates. Removal of zinc from thermolysin abolishes the esterase activity of the native enzyme. Among the metals examined, only Co2+ and Zn2+ restore esterase activity to any significant extent, Co2+ restoring 50% and Zn2+ 100% of the native thermolysin activity. The hydrolysis of esters and peptides by thermolysin does not differ with respect to either the binding or catalytic steps. Substrate specificity, pH-rate profiles, inhibitor, and deuterium isotope effects are identical for both types of substrates.

show abstract

Section: Discussionmentioning

confidence: 99%

Esterase activity of zinc neutral proteases

Holmquist¹,

Vallée²

1976

Biochemistry

View full text Add to dashboard Cite

show abstract

“…With many bacterial proteases stabilizing effect of CaH has been reported. 33 10, the four of alkaline-earth metal cations were tested and only Ca H could stabilize the Sfericase at high temperature. These results were quite different from those reported by McConn and Miyata.…”

Section: Table V Effect Of Some Cations For Edta Inactivation At Low Temperaturementioning

confidence: 99%

Purification and Some Properties ofBacillus sphaericusProtease

Yoshida¹,

H²,

Miyado³

et al. 1977

Agricultural and Biological Chemistry

View full text Add to dashboard Cite

A new protease, Sfericase, found in culture filtrate of a strain of Bacillus sphaericus was purified and isolated in a crystalline form by salting out with ammonium sulfate followed by column chromatography on DEAE-Sephadex A-50 (using a specific elution buffer). The optimal and maximal caseinolytic activities of the protease were around pH 9.0~9.3 and at 55°C, respectively. The proteolytic activity was inhibited by DFP, potato protease inhibitors and EDTA, but not by PCMB, o-phenanthroline, TPCK and TLCK. The molecular weight of the protease was estimated by sedimentation equilibrium (32,000±400) and gel chromatography (27,000).The enzyme molecule contained two atoms of undialyzable calcium ion and two halfcystine residues which should afford one disulfide linkage as no free sulfhydryl group was found even with the denatured enzyme.By immunological cross reaction, it was indicated that the enzyme was distinct from other alkaline proteases of Bacillus origins.

show abstract

Proteases of the genus Bacillus. I. Neutral proteases

Keay

Wildi

1970

Biotech & Bioengineering

140

View full text Add to dashboard Cite

B. subtdis NRRL B3411 neutral protease has been extensively purified by solvent and salt fractionation, pigment removal with DEAE-cellulose followed by chromatography on hydroxylapatite, and a final passage through a Sephadex G-100 column. The neutral protease was shown to be homogeneous by disc gel and cellulose acetate electrophoresis, gel filtration chromatography, and ultracentrifugation. The molecular weight w&s determined by osmometry and ultracentrifugation to be about 3842,000 and the amino acid composition and zinc content determined. The general properties of the enzyme, pH-activity relationship, stability, effect of inhibitors, and specificity are discussed. Comparative studies were carried out on the B. subtilas NRRL B3411 and B. sublilis vur. umylosacchuriticus neutral proteases and these enzymes were found to be indistinguishable by the methods used, but quite distinct from the thermostable enzyme thermolysin from B. thermoproteolyticus. ABBREVLATIONS FAGLA-Furylacryloylglycyl Gleucine amide CBZ -Carbobenzoxy DFP -DiisopropyHuorophosphate DEAE -Diethylaminoethyl CM-carboxymethyl P N E -p-nitrophenyl ester TMED -tetramethylethylenediamine 179 possessing similar specificities with respect to the peptide bonds cleaved2J; and (c) alkaline proteases having maximum activity at pH 9.0-11.0, cleaving a wide range of peptide bonds, possessing esterase activity, and unaffected by metal-chelating agents but inhibited by DFP.4,5Microbial enzymes have recently found widespread application in the field of detergents and so far all of the enzymes in use are produced by organisms of the genus Bacillus. The neutral protease may not play as significant a role in detergent applications of the enzymes as the alkaline protease because of the high pH in detergent solutions as well as the relatively poor stability of the neutral protease.6 B. subtilis enzymes have also been shown to reduce dental plaque in humans? and the neutral protease is the more effective agent in this application.The neutral proteases have been less extensively studied than the alkaline proteases. The neutral protease of B. subtilis var. amylo-~a c c h a r i t i c u s~.~ is claimed to be different from the neutral protease of a strain of B subtilis (now registered as strain S R R L B3411)6J0J1 in certain properties. The protease of B. megaterium shows similar general properties and specificity to the B. subtilis neutral protease.12J3The other Bacillus neutral protease which has been studied in any depth is thermolysin, produced by B. thermoproteolyticus, a strain of B. ~tearothermophilus.~~ Thermolysin is quite different from the other neutral proteases having very high thermostability and amino acid composition,15 although its specificity of bond cleavage on both synthetic peptides and B-chain of insulin is ~i r n i l a r .~.~~ A preliminary account of the distinctions between the B. subtilis and B. thermoproteolyticus enzymes has been published.'? This paper gives a detailed account of the isolation of B. subtilis neutral protease by a new method,...

show abstract

Studies on Bacterial Protease

Cited by 22 publications

References 3 publications

Esterase activity of zinc neutral proteases

Esterase activity of zinc neutral proteases

Purification and Some Properties ofBacillus sphaericusProtease

Proteases of the genus Bacillus. I. Neutral proteases

Contact Info

Product

Resources

About