Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance

Monteiro, Paulo Sérgio; Guimarães, Valéria Monteze; Melo, Ricardo Rodrigues de; Rezende, Sebastião Tavares de

doi:10.1590/s1517-838220120037

Cited by 25 publications

(32 citation statements)

References 28 publications

(55 reference statements)

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“…Molecular weight of PASN03 (70.87 kDa), PASN11 (72.54 kDa), and PASN12 (57.46 kDa) phytases were compareable with weights of phytases from A. ficuum NTG-23 (65.5 kDa) (Zhang et al, 2010) and A. niger CFR 335 (66kDa) (Gunashree and Venkateswaran, 2015). Molecular weight (87.41kDa) of phytase of PASN10 was comparable with molecular weight of A. niger NICM (87kDa) (Bhavsar et al, 2011) and A. niger UFV-1 (81kDa) (Monteiro et al, 2015). Molecular weights PASN04, PASN06 and PASN08 were 100.54, 107.82 and 105.34 kDa, respectively which were similar to the molecular weights of phytase from A. niger FS3 (108kDa) (Spier et al, 2011) and R. oligospora (120kDa) (Casey and Walsh, 2004).…”

Section: Molecular Weight Of Phytasementioning

confidence: 67%

“…Isolates were confirmed as positive for phytase if the zone of hydrolysis was retained. Phytase production: Phytase was produced following the method of Monteiro et al (2015). Phytase was produced by inoculating ~10 6 spores of A. niger in 250mL flasks containing sterilized phytase screening broth (pH 5.6).…”

Section: Methodsmentioning

confidence: 99%

“…In present study, phytase activities of cell free supernatant of A. niger isolates grown in PSM broth ranged from 68.88±2.55 to 274.99±10.14 FTU/mL which were comparable with previous reports. Monteiro et al (2015) analyzed phytase production by A. niger UFV-1 and reported that it had 138.6 U/mL phytase enzyme activity. In another previous study by Rani and Ghosh (2011), phytase enzyme has been reported form Aspergillus oryzae.…”

Section: Identification Of Fungimentioning

confidence: 99%

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PRODUCTION AND CHARACTERIZATION OF PHYTASE FROM INDIGENOUS Aspergillus niger ISOLATES

Tariq¹

2017

PAKJAS

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Aim of the present study was to isolate phytase producing Aspergillus niger, and production, purification and characterization of phytase enzyme which is responsible for conversion of organic phosphorous (phytate) into inorganic form (available). Indigenous A. niger isolates (n=20) were identified through macroscopic (obverse side: brown to black, granular, folded, cottony from center with smooth and white edges, reverse side: center light pale, opaque edges and ridges) and microscopic characteristics (phialospores in chains, circular vesical, metullae, phialids, septate and hyaline hyphae). Isolates were screened for their ability to produce phytase on Phytase Screening Medium. Nine isolates (45%) gave positive screening results through cobalt chloride staining (2% cobalt chloride, 6.25% ammonium molybdate and 0.42% ammonium vanadate). Diameter of colonies, zone of hydrolysis and zone of hydrolysis to colony diameter ratio were 20.97-43.00 mm, 25.03-49.00 mm and 1.13-1.19, respectively. Phytase activity of cell free supernatants of indigenous A. niger isolates ranged from 68.88±2.55 to 274.99±10.14 FTU/mL. PASN01 and PASN06 exhibited highest enzyme activity (274.99±10.14 and 274.99±9.00 FTU/mL, respectively). Phytase were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Molecular weights of phytases from A. niger isolates ranged from 35.21 to 107.82 kilo Daltons. It is concluded that indigenous A. niger PASN01 and PASN06 may have commercial application after further characterization.

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Section: Molecular Weight Of Phytasementioning

confidence: 67%

Section: Methodsmentioning

confidence: 99%

Section: Identification Of Fungimentioning

confidence: 99%

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PRODUCTION AND CHARACTERIZATION OF PHYTASE FROM INDIGENOUS Aspergillus niger ISOLATES

Tariq¹

2017

PAKJAS

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“…Phytase activity was determined by measuring the Pi released from sodium phytate solution (Harland and Hraland, 1980;Monteiro et al, 2015). The reaction mixture consisted of 1 mL of 0.1 M MgSO4*7H2O, 2.4 mL of 6.82 mM phytic acid and 0.6 mL of appropriately diluted crude enzyme solution.…”

Section: Enzyme Activity Assaymentioning

confidence: 99%

Optimized production of phytase by solid-state fermentation using triticale as substrate and source of inducer

Neira-Vielma¹,

Cristobal²,

Anna³

et al. 2018

Afr. J. Biotechnol.

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This study was carried out to evaluate the process of phytase production by Aspergillus niger in solidstate fermentation (SSF) using triticale waste. A waste that currently has no use was reported for this biotechnological process, and is of high impact due to the null use. The process was carried out using an additive free medium, supplemented with only one nitrogen source. Under these conditions, phytase activity of 7.45 U/g dry substrate (DS) was obtained. The process was optimized using different additives such as dextrose, lactose, Tween 80 and potassium chloride. For fermentation maximization, two experimental designs were used: 1) Plackett-Burman design (PBD) and 2) the Box-Behnken design (BBD). PBD was used to evaluate the effect of related variables on the production of phytase, as well as their level of significance in the process, while BBD was used for optimal conditions determination. The process was conducted with Petri dishes and a maximum enzyme activity of 25.8 IU/g DS was obtained. Subsequently, SSF was carried out in a tray to increase the amount of fermented substrate and phytase activity of 23.63 IU/g DS was obtained. The results of this study suggest a minimal decrease (8.4%) in enzyme production with scaling.

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“…Fitase obtida do fungo A. niger e expressa em P. pastoris mostrou atividade máxima aos 55°C (ZHAO et al, 2007). Fitases de A. flavus e A. niger UFV-1, apresentaram temperaturas ótimas de 45 e 50°C, respectivamente (GAIND;MONTEIRO et al, 2015). Fitases de Candida krusei Wz-001, R. microsporus e A. fumigatus tiveram atividade máxima em 40 °C (QUAN et al, 2002;SATO, 2015;SANNI;LAWAL;ENUJIUGHA, 2019) e uma fitase de A. niger CFR 335, apresentou 100% de atividade em temperatura de 30°C (GUNASHREE; VENKATESWARAN, 2015).…”

Section: Dicroismo Circular (Cd)unclassified

Clonagem, expressão e caracterização de fitase do fungo termofílico >i<Myceliophthora thermophila>/i<

Ferreira¹

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Fitases são enzimas que hidrolisam o ácido fítico, substâncias orgânicas complexas amplamente encontradas na natureza, principalmente em cereais e leguminosas. Nesses alimentos vegetais, o fitato é a principal fonte de fósforo, correspondendo entre cinquenta a oitenta por cento do conteúdo de fósforo orgânico. Entretanto, o fósforo do fitato é muito pouco utilizado por humanos e outros animais monogástricos, como suínos e aves, devido à ausência ou insuficiência das enzimas que o degradam. Assim, rações contêm o fósforo orgânico do fitato e fósforo inorgânico adicionado, que é um mineral caro e não renovável. Em áreas de criação intensiva o excesso de fósforo é excretado no ambiente, causando problemas, tais como a eutrofização. Devido aos problemas ambientais e ao alto custo do fósforo inorgânico, a suplementação de enzimas, principalmente fitases, tem ganhado destaque, entre elas, as enzimas fúngicas, devido a sua estabilidade a diferentes temperaturas e pH. Além disso, técnicas de engenharia genética permitem a transferência de genes entre espécies, resultando em muitos benefícios, principalmente em relação ao aumento e melhoramento na produção de enzimas industriais. O objetivo deste trabalho foi realizar a clonagem, expressão heteróloga, purificação e caracterização bioquímica da enzima fitase (MtPhy) obtida a partir do fungo termofílico M. thermophila. O gene que codifica a enzima foi clonado no vetor pEXPYR e heterologamente expresso em A. nidulans. A proteína recombinante foi purificada e teve sua identidade confirmada por espectrometria de massas. Nas análises bioquímicas, apresentou pH ótimo de 5,0 e temperatura ótima de 60°C e manteve praticamente 100% de sua atividade a 50°C por 2 horas e pela análise de dicroísmo circular e ThermoFluor, apresentou desenovelamento de sua estrutura nas temperaturas de 63 e 67°C, respectivamente. Demonstrou atividade aumentada na presença de CaCl 2 e MgCl 2 e foi fortemente inibida por FeCl 3 , CuSO 3 e SDS. Sua atividade específica (954,8 U/mg) utilizando fitato de sódio foi bastante superior a de outras fitases descritas na literatura. Demonstrou-se efetiva na liberação do fósforo em rações e extremamente resistente às proteases, tripsina e pepsina, presentes no trato digestório de animais monogástricos, demonstrando dessa forma, seu elevado potencial para aplicação industrial. Palavras-chave: Fitases. Fungos termofílicos. Myceliophtora thermophila.

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Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance

Cited by 25 publications

References 28 publications

PRODUCTION AND CHARACTERIZATION OF PHYTASE FROM INDIGENOUS Aspergillus niger ISOLATES

PRODUCTION AND CHARACTERIZATION OF PHYTASE FROM INDIGENOUS Aspergillus niger ISOLATES

Optimized production of phytase by solid-state fermentation using triticale as substrate and source of inducer

Clonagem, expressão e caracterização de fitase do fungo termofílico >i<Myceliophthora thermophila>/i<

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