Purification and Properties of a Protease Produced by Bacillus Subtilis CN2 Isolated from a Vietnamese Fish Sauce

Uchida, Hiroyuki; Kondo, Daisaku; Yamashita, Satoko; Tanaka, Tomoko; Tran, Lien Ha; Nagano, Hiroko; Uwajima, Takayuki

doi:10.1023/b:wibi.0000043168.15067.44

Cited by 36 publications

(30 citation statements)

References 12 publications

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“…Proteases with distinct optimal values of pH have been reported, and the value presented in this paper describes proteases that are active at a near-neutral pH. The data presented are similar to those of the alkaline proteases from Bacillus subtilis CN2 reported in the pH range of 7-11 (Uchida et al, 2004). Bolumar et al (2005) showed that neutralalkaline proteases ranging in pH 6.0 to 12, showed beat around pH 8.0 with stability at temperatures above 75ºC, while there was decrease of stability at 37°C.…”

Section: Enzyme Assay For Extracellular Proteolytic Enzyme Productionsupporting

confidence: 72%

Effect of pH and temperature on the production and activity of Schwanniomyces polymorphus extracellular proteases in fermentation medium

Alexander¹,

Cleiton²,

Dangely³

et al. 2015

Afr. J. Microbiol. Res.

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Section: Enzyme Assay For Extracellular Proteolytic Enzyme Productionsupporting

confidence: 72%

Effect of pH and temperature on the production and activity of Schwanniomyces polymorphus extracellular proteases in fermentation medium

Alexander¹,

Cleiton²,

Dangely³

et al. 2015

Afr. J. Microbiol. Res.

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“…An optimum pH of 11 of alkaline protease was observed in the protease produced by Bacillus sp 41,42 .…”

Section: Optimum Reaction Temperature and Phmentioning

confidence: 94%

Production of an alkaline protease using Bacillus pumilus D3 without inactivation by SDS, its characterization and purification

Özçelik

Aytar

Gedikli

et al. 2013

Journal of Enzyme Inhibition and Medicinal Chemistry

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In this study, protease-producing capacity of Bacillus pumilus D3, isolated from hydrocarbon contaminated soil, was evaluated and optimized. Optimum growing conditions for B. pumilus D3 in terms of protease production were determined as 1% optimum inoculum size, 35 C temperature, 11 pH and 48 h incubation time, respectively. Stability studies indicated that the mentioned protease was stable within the pH range of 7-10.5 and between 30 C and 40 C temperatures. Surprisingly, the activity of the enzyme increased in the presence of SDS with concentration up to 5 mM. The protease was concentrated 1.6-fold with ammonium sulfate precipitation and dialysis. At least six protein bands were obtained from dialysate by electrophoresis. Four clear protein bands with caseinolytic activity were detected by zymography. Dialysate was further purified by anion-exchange chromatography and the caseinolytic active fraction showed a single band between 29 and 36 kDa of reducing conditions.

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“…Nascimento 16 also found an alkaline protease which was active in a broad pH range of 6 to 10, with a relative activity of 60% retained at pH above 9. The pH stability of protease produced from B. subtilis PE-11 6 and B. subtilis CN2 15 were found to be in the ranges of 8 to 11 and 7 to 11 with the relative activity of more than 90% and 70%, respectively. Other strong inhibitors were found to be PCMB, EDTA, PMSF and 1, 10 Phenanthroline, with resulted relative activity of 2, 7, 7 and 12%, respectively.…”

Section: Inhibitors Of the Protease Inhibitors Of The Protease Inhibimentioning

confidence: 98%

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Yossan¹,

Reungsang²,

Yasuda³

2006

ScienceAsia

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An alkaline protease produced by Bacillus megaterium isolated from a fermented broth of Thai fish sauce was purified by hydrophobic interaction combined with gel filtration techniques. After final purification step, the enzyme was purified 148-fold with an increase in specific activity from 0.09 to 13.33 U/mg protein.The properties of the purified enzyme were then analyzed. The molecular weight of the enzyme under denaturing condition was estimated to be 27 kDa. The optimum pH and temperature for protease activity were 10 and 50 o C, respectively. This protease could retain the activity in the pH and temperature ranging from pH 7.5 to 9.5 and 30 to 45 o C, respectively, resulting in the relative activity of higher than 80%. The enzyme was completely inhibited by diisopropyl fluorophosphate (DFP) suggesting that it was a serine protease. Phenylmethylsulfonyl fluoride (PMSF), p-Chloromercuribenzoic acid (PCMB), ethylenediaminetetraacetic acid (EDTA), 1, 10-phenanthroline and Fe 3+ strongly inhibited the activity of this purified enzyme activity, decreasing relative activity to lower than 15%. Protease activity was enhanced by Mn 2+ , Ca 2+ and Mg 2+ . Cytochrome C, soybean protein isolate and casein were good substrates specific to the enzyme with the relative activity of more than 100%.

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Purification and Properties of a Protease Produced by Bacillus Subtilis CN2 Isolated from a Vietnamese Fish Sauce

Cited by 36 publications

References 12 publications

Effect of pH and temperature on the production and activity of Schwanniomyces polymorphus extracellular proteases in fermentation medium

Effect of pH and temperature on the production and activity of Schwanniomyces polymorphus extracellular proteases in fermentation medium

Production of an alkaline protease using Bacillus pumilus D3 without inactivation by SDS, its characterization and purification

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