An alkaline proteinase of an alkalophilicBacillus sp.

Tsuchida, Osamu; Yamagata, Yuriko; Ishizuka, Takehiko; Arai, Takeji; Yamada, Jun−ichi; Takeuchi, Michio; Ichishima, Eiji

doi:10.1007/bf01568094

Cited by 116 publications

(63 citation statements)

References 8 publications

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“…Relative enzyme activity by different inhibitors is described in Figure 8. Our findings are similar to those of Tsuchida et al (1986) where the protease was completely inhibited by PMSF. This indicated that it is a serine alkaline protease.…”

Section: Effect Of Protease Inhibitors On Enzyme Activitysupporting

confidence: 90%

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Isolation, purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus thuringinsis-SH-II-1A

Harer¹,

Bhatia²,

Bhatia³

2018

Afr. J. Biotechnol.

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In the present study, the isolation, purification and partial characterization of thermostable serine alkaline protease produced from Bacillus thuringinsis SH-II-1A was reported. The culture was isolated from soil of slaughter house waste and identified further from ribosomal sequence. The crude enzyme was purified by ammonium sulfate precipitation, dialysis and Sephadex G-200 gel permeation chromatography up to 17.04 fold with recovery of 8.47%. Relative molecular weight (67 kDa) of purified enzyme was observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Maximum production of enzyme and cell biomass was observed for 48 h of incubation period at 45°C. Strong activity of enzyme was observed at pH 10 to 11; also stability of up to 2 and 20 h incubation at the same pH range confirms alkaline protease. Optimum temperature recorded for protease activity was 45°C, and 100% thermal stability up to 350 min of incubation was recorded. Among different natural substrates tried, casein was found as ideal substrate. Enzyme activity was strongly enhanced by metal ions like Ca 2+ , Mg 2+ and Mn 2+ whereas, 100% enzyme activity was inhibited by phenylmethylsulphonyl fluoride (PMSF), and up to 92% inhibition by diisopropyl fluorophosphates (DFP) confirmed serine protease. Detergent compatibility of the enzyme was studied in the presence of 10 mM CaCl 2 and 1 M glycine at 45°C. This indicates 80 to 100% stability for a period of 0.5 to 2.5 h incubation. Improved washing performance and removal of blood stains from the cotton cloth was observed when detergent Surf excel was used with enzyme. Overall, the observed properties of isolated protease conclude its commercial application in detergent and leather industries.

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Section: Effect Of Protease Inhibitors On Enzyme Activitysupporting

confidence: 90%

“…Caseinase activity was assessed by the modified procedure (Tsuchida et al, 1986) using 2% casein substrate dissolved in 0.2 M carbonate buffer pH 10. Casein solution (0.5 ml) with an equal volume of suitably diluted enzyme solution was incubated at 37°C.…”

Section: Determination Of Proteolytic Activity (Caseinase Determination)mentioning

confidence: 99%

Isolation, purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus thuringinsis-SH-II-1A

Harer¹,

Bhatia²,

Bhatia³

2018

Afr. J. Biotechnol.

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“…Protease activity in the crude enzyme was assayed by the modified Tsuchida procedure [12] using 1% casein as substrate. Casein solution (1.0 ml) was mixed with an equal volume of crude protease enzyme solution and incubated at 45˚C for 10 min.…”

Section: Assay Of Enzyme Activitymentioning

confidence: 99%

Enzymatic Decolorization of Malachite Green Dye by a Newly Isolated Bacillus Cereus Strain wwcp

Wanyonyi¹,

Onyari²,

Shiundu³

et al. 2014

IOSRJESTFT

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“…Protease activity in the culture filtrate of S. griseolus was assayed by method of Tsuchida et al [28] by using casein as the substrate. One protease unit is defined as the amount of enzyme that releases 1µg of a tyrosine per ml per minute under the above assay conditions.…”

Section: Quantitative Assay Of Proteasementioning

confidence: 99%

In Vitro Action of Streptomyces griseolus Proteases as Bio-Control on Fasciola gigantic Eggs

Gammal¹,

Shalaby²

2014

J Bacteriol Parasitol

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This work evaluated the in vitro bio-control action of actinomycetes isolated from Egyptian soil against Fasciola gigantica eggs. In this respect, the identified Streptomyces griseolus was the best parasite-control agent which gave the highest mortality percent applying three methods; the first method was the application of the grown bacterial disks with the eggs at zero time on the medium (2% w/v water-agar) and incubated for 21 days (75.5% mortality), the second method was the incubation of the bacterial disks using the same medium for 5 days before eggs inoculation, after 21 days of incubation the mortality percent was 80.8%. The application of the third method showed a good effect by using all bacterial culture filtrates. S. griseolus filtrate which had the highest lytic enzyme and proteolytic activities reached 2.0 and 660.7 U/ml, respectively, gave the highest mortality (95.5%) of F. gigantica eggs compared to the other bacterial filtrates. The crude culture filtrate and the diluted culture filtrate (1.33 fold) of S. griseolus gave the highest mortality percent against the parasite´s eggs (95.8 and 94.5%, respectively) compared to the higher diluted filtrates. The culture filtrate of S. griseolus grown on two different media and its partially purified enzyme (precipitated by 60% ammonium sulfate) were tested for their proteolytic activity and inhibitory effect against F. gigantica eggs. Medium no. 2 showed the highest proteolytic activity (1030.3 and 1138.1 U/ml) for the crude and the partially purified enzyme, respectively. While the highest inhibitory effect reached 97.6 and 89.9%, respectively. Finally, the application of crude enzyme filtrate was better than the partially purified one. Streptomyces griseolus was proved as a potential biological control agent for this helminth.

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An alkaline proteinase of an alkalophilicBacillus sp.

Cited by 116 publications

References 8 publications

Isolation, purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus thuringinsis-SH-II-1A

Isolation, purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus thuringinsis-SH-II-1A

Enzymatic Decolorization of Malachite Green Dye by a Newly Isolated Bacillus Cereus Strain wwcp

In Vitro Action of Streptomyces griseolus Proteases as Bio-Control on Fasciola gigantic Eggs

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