A statistical approach for the enhanced production of alkaline protease showing fibrinolytic activity from a newly isolated Gram-negative Bacillus sp. strain AS-S20-I

Mukherjee, Ashis K.; Sudhir, K.

doi:10.1016/j.nbt.2010.11.003

Cited by 65 publications

(36 citation statements)

References 29 publications

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“…(Streptomycetaceae) isolated from Amazonian lichens ACTA AMAZONICA Proteolytic activity significantly was inhibited by PMSF, an inhibitor of serine proteases, which indicates that the enzyme's catalytic site includes a serine residue. These results are in agreement with (Afifah et al 2014;Mukherjee and Rai 2011).…”

Section: Discussionsupporting

confidence: 92%

Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens

et al. 2016

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Thrombosis is a pathophysiological disorder caused by accumulation of fibrin in the blood. Fibrinolytic proteases with potent thrombolytic activity have been produced by diverse microbial sources. Considering the microbial biodiversity of the Amazon region, this study aimed at the screening, production and biochemical characterization of a fibrinolytic enzyme produced by Streptomyces sp. isolated from Amazonian lichens. The strain Streptomyces DPUA1576 showed the highest fibrinolytic activity, which was 283 mm 2 . Three variables at two levels were used to assess their effects on the fibrinolytic production. The parameters studied were agitation (0.28 -1.12 g), temperature (28 -36 ºC) and pH (6.0 -8.0); all of them had significant effects on the fibrinolytic production. The maximum fibrinolytic activity (304 mm 2 ) was observed at 1.12 g, 28 ºC, and pH of 8.0. The crude extract of the fermentation broth was used to assess the biochemical properties of the enzyme. Protease and fibrinolytic activities were stable during 6 h, at a pH ranging from 6.8 to 8.4 and 5.8 to 9.2, respectively. Optimum temperature for protease activity ranged between 35 and 55 °C, while the highest fibrinolytic activity was observed at 45 ºC. Proteolytic activity was inhibited by Cu 2+ and Co 2+ ions, phenylmethylsulfonyl fluoride (PMSF) and pepstatin A, which suggests that the enzyme is a serine protease. Enzymatic extract cleaved fibrinogen at the subunits Aα-chain, Aβ-chain, and γ-chain. The results indicated that Streptomyces sp. DPUA 1576 produces enzymes with fibrinolytic and fibrinogenolytic activity, enzymes with an important application in the pharmaceutical industry. KEYWORDS: Actinomycetes, fibrinolytic protease, inhibitor protease, fibrinogenolytic activity.Seleção, produção e caracterização bioquímica de uma nova enzima fibrinolítica produzida por Streptomyces sp. (Streptomycetaceae) isolada de liquens da Amazônia RESUMO A trombose é uma doença patofisiológica causada pelo acúmulo de fibrina no sangue. Proteases fibrinolíticas com potente atividade trombolítica são produzidas por diversas fontes microbianas. Considerando a biodiversidade microbiana da região amazônica, o presente estudo teve como objetivo a seleção, produção e caracterização bioquímica da enzima fibrinolítica de Streptomyces sp. isolado de líquens da Amazônia. Streptomyces DPUA1576 foi a melhor produtora com atividade fibrinolítica de 283 mm 2 . Três variáveis em dois níveis foram utilizadas para determinar as variáveis mais relevantes na produção da enzima fibrinolítica (FA). Os parâmetros estudados foram agitação (0.28 -1.12 g), temperatura (28 -36 ºC) e pH (6.0 -8.0) e todos obtiveram efeitos significativos na produção fibrinolítica. A maior atividade fibrinolítica (304 mm 2 ) foi obtida a 1.12 g, 28 ºC e pH 8.0. O extrato bruto da fermentação foi usado para determinar as propriedades bioquímicas da enzima. Atividades proteásica e fibrinolítica foram estáveis durante 6 horas no intervalo de pH entre 6.8 -8.4 e 5.8 -9.2, respectivamente. Temperatura ...

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Section: Discussionsupporting

confidence: 92%

Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens

et al. 2016

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“…The fibrinolytic enzyme yield obtained in this study was found to be higher than the results reported on fibrinolytic enzyme production by Bacillus sp. (Mukherjee and Rai, 2011) and B. vallismortis Ace02 (Kim et al, 2007). B. cereus SRM-001 isolate used in this study may be used for the enhanced production of fibrinolytic enzyme for medical applications.…”

Section: Discussionmentioning

confidence: 99%

Fibrinolytic enzyme production by newly isolated <i>Bacillus cereus</i> SRM-001 with enhanced <i>in-vitro</i> blood clot lysis potential

Narasimhan

Chandrasekaran²,

Rajesh

2015

J. Gen. Appl. Microbiol.

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The discovery of plasmin-like microbial fibrinolytic enzymes having high specificity and negligible side effects is crucial for thrombolytic therapy. Herein, we report one such extra-cellular fibrinolytic enzyme producing Bacillus cereus SRM-001 isolated from the blood-laden soil of a chicken dump yard. The potency of the enzyme was established with fibrin plate assay and in-vitro blood clot lysis assay. The shake-flask operating parameters and media composition were optimized for maximizing the productivity of the enzyme. The operating parameters, pH 7, 37°C, 1% inoculum volume and 24 h inoculum age, were found to be the optimum. The levels of media components, corn flour (0.3% w/v), soyabean powder (1.9% w/v) and MnSO 4 (11.5 mM) were optimized by statistical analysis using Box-Behnken design derived RSM. This resulted in an almost 1.8 fold increase in fibrinolytic enzyme productivity. The 3D response surface plots showed soyabean powder and MnSO 4 to be the key ingredients for enhancing the enzyme productivity, whereas corn flour had a marginal effect. The in-vitro blood clot lysis assay conducted at near physiological pH 7 at 37°C showed the enzyme to be a potential therapeutic thrombolytic agent.

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“…Furthermore, microbial enzyme production always favours the economy of bulk production (Bajaj et al 2013). Most of the studies on fibrinolytic proteases involved purification and characterization of enzymes (Mahajan et al 2012) with relatively a fewer reports on optimization of enzyme production (Mukherjee and Rai 2011). Literature shows that studies on fungal fibrinolytic proteases have been more emphasized than bacterial ones (Simkhada et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Optimization of fibrinolytic protease production from Bacillus subtilis I-2 using agro-residues

Bajaj¹,

Singh²,

Khullar³

et al. 2014

Braz. arch. biol. technol.

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A statistical approach for the enhanced production of alkaline protease showing fibrinolytic activity from a newly isolated Gram-negative Bacillus sp. strain AS-S20-I

Cited by 65 publications

References 29 publications

Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens

Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens

Fibrinolytic enzyme production by newly isolated <i>Bacillus cereus</i> SRM-001 with enhanced <i>in-vitro</i> blood clot lysis potential

Optimization of fibrinolytic protease production from Bacillus subtilis I-2 using agro-residues

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