Purification and characterization of a novel Aspergillus heteromorphus URM 0269 protease extracted by aqueous two-phase systems PEG/citrate

Fernandes, Lígia Maria Gonçalves; Carneiro-da-Cunha, Márcia Nieves; Silva, Jônatas de Carvalho; Porto, Ana Lúcia Figueiredo; Porto, Tatiana Souza

doi:10.1016/j.molliq.2020.113957

Cited by 16 publications

(7 citation statements)

References 40 publications

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“…In a previous study by our research group, the protease obtained by SSF using the same fungal strain and substrate (wheat bran) showed different optimal conditions (40 • C and pH 8.0) [19], suggesting that the fermentation method may directly influence the characteristics of the biocatalyst. A similar optimum temperature was observed for proteases obtained from different Aspergillus species, such as A. heteromorphus [20], A. niger [21] and A. oryzae [22], in conventional fermentation processes (SSF and SmF).…”

Section: Effect Of Temperature and Ph On Protease Activitysupporting

confidence: 69%

“…The relatively low ∆H* value (37.90 kJ mol −1 ) indicates that the formation of the transition state or activated enzyme-substrate complex occurred effectively; however, this value is higher than that reported by Fernandes et al [20] for azocasein hydrolysis by A. heteromorphus protease (21.8 kJ mol −1 ). As known, ∆S* is correlated to the order degree of a reaction system; therefore, in enzyme-catalyzed reactions negative values such as that estimated in this study (−73.94 J K −1 mol −1 ) suggest that the structure of enzyme-substrate at transition state is more ordered than that of the enzyme-substrate complex.…”

Section: Kinetic and Thermodynamic Parameters Of Azocasein Hydrolysiscontrasting

confidence: 55%

“…These results indicate that the protease produced by SF is more thermostable than that obtained by SSF; therefore, one of the next efforts will be an enzyme structural analysis that can provide some additional information for a better understanding of the thermostabilization mechanism under SF conditions. Moreover, the protease produced in the present study appears to be more thermostable than other Aspergillus proteases at same temperature (50 • C), such as those produced by A. fumigatus (t 1/2 : 96.3 min) [28], A. niger (t 1/2 : 135.91 min; D-value: 451.49 min) [30] and A. heteromorphus (t 1/2 : 54.6 min; D-value: 181.3 min) [20].…”

Section: Kinetic and Thermodynamic Parameters Of Protease Thermal Denaturationmentioning

confidence: 88%

“…As known, ∆G* is the most suitable thermodynamic parameter to measure the feasibility and extent of a reaction, as the lower the ∆G* value the more feasible the reaction, i.e., the conversion of the reactant into the product is more spontaneous [35]. The value of ∆G* estimated for azocasein hydrolysis catalyzed by A. tamarii URM4634 protease (59.94 kJ mol −1 ) is lower than those of casein and azocasein hydrolysis catalyzed by other fungal (A. heteromorphus: ∆G* = 65.8 kJ mol −1 [20]) and bacterial (Bacillus stearothermophilus: ∆G* = 91.71 kJ mol −1 [32]; Bacillus cereus: ∆G* = 68.68-69.42 kJ mol −1 [36]) proteases, which indicates that the conversion of the transition state of enzyme-substrate complex into the product is more spontaneous.…”

Section: Kinetic and Thermodynamic Parameters Of Azocasein Hydrolysismentioning

confidence: 89%

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Use of a Sequential Fermentation Method for the Production of Aspergillus tamarii URM4634 Protease and a Kinetic/Thermodynamic Study of the Enzyme

Oliveira¹,

Claudino²,

Converti

et al. 2021

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Microbial proteases are commonly produced by submerged (SmF) or solid-state fermentation (SSF), whose combination results in an unconventional method, called sequential fermentation (SF), which has already been used only to produce cellulolytic enzymes. In this context, the aim of the present study was the development of a novel SF method for protease production using wheat bran as a substrate. Moreover, the kinetic and thermodynamic parameters of azocasein hydrolysis were estimated, thus providing a greater understanding of the catalytic reaction. In SF, an approximately 9-fold increase in protease activity was observed compared to the conventional SmF method. Optimization of glucose concentration and medium volume by statistical means allowed us to achieve a maximum protease activity of 180.17 U mL−1. The obtained enzyme had an optimum pH and temperature of 7.0 and 50 °C, respectively. Kinetic and thermodynamic parameters highlighted that such a neutral protease is satisfactorily thermostable at 50 °C, a temperature commonly used in many applications in the food industry. The results obtained suggested not only that SF could be a promising alternative to produce proteases, but also that it could be adapted to produce several other enzymes.

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Section: Effect Of Temperature and Ph On Protease Activitysupporting

confidence: 69%

Section: Kinetic and Thermodynamic Parameters Of Azocasein Hydrolysiscontrasting

confidence: 55%

Section: Kinetic and Thermodynamic Parameters Of Protease Thermal Denaturationmentioning

confidence: 88%

Section: Kinetic and Thermodynamic Parameters Of Azocasein Hydrolysismentioning

confidence: 89%

See 2 more Smart Citations

Use of a Sequential Fermentation Method for the Production of Aspergillus tamarii URM4634 Protease and a Kinetic/Thermodynamic Study of the Enzyme

Oliveira¹,

Claudino²,

Converti

et al. 2021

Catalysts

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“…Um estudo realizado porFernandes et al (2020) para purificar proteases de Aspergillus heteromorphus produzidas por fermentação em estado sólido apresentou pH ótimo 8, semelhante ao apresentado no presente trabalho. Semelhantemente os autores também observaram a partição preferencial da protease para fase PEG e observaram efeito positivo da concentração de citrato e negativo da massa molar do PEG para o K. Porém a massa molar do PEG maior (8000 g/mol) foi melhor para purificação da enzima.…”

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PURIFICAÇÃO DE PROTEASES DE Aspergillus heteromorphus URM 0269 POR SISTEMA DE DUAS FASES AQUOSAS PEG-Citrato

Menezes¹,

Silva²,

de³

et al. 2021

Inovação Gestão E Sustentabilidade Na Agroindústria

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As proteases são enzimas capazes de catalisar a hidrólise de ligações peptídicas e apresentam diversas aplicações biotecnológicas, sendo uma das principais enzimas empregadas nos diferentes setores industriais, especialmente na indústria de alimentos. A produção dessas enzimas através de métodos fermentativos apresenta diversas vantagens como baixo custo, rendimento elevado e produção em larga escala, porém, a presença de impurezas nos extratos obtidos no final do processo pode ocasionar diminuição no potencial biotecnológico em termos de aplicações, sendo assim, necessário técnicas que proporcionem a viabilidade da função e estrutura das proteases. O Sistema de Duas Fases Aquosas (SDFA) é uma técnica simples, econômica e com elevando rendimento de extração. Diante disso, o SDFA PEG/Citrato foi utilizado para extrair e purificar parcialmente as proteases produzidas a partir do micro-organismo Aspergillus heteromorphus URM 0269 por fermentação submersa (FSm). Para essa finalidade, foi realizado um planejamento fatorial completo 2 4 tendo como variáveis reposta o coeficiente de partição (K), recuperação (Y) e fator de purificação (FP) e as variáveis independentes estudadas foram a massa molar de PEG (MPEG), concentração de PEG (CPEG), concentração de citrato (CCITRATO) e pH. A protease apresentou maior preferência pela fase superior, rica em PEG, onde as variáveis MPEG e CCITRATO, bem como a interação entre ambas, apresentaram efeitos significativos para todas as variáveis resposta. O ensaio 5 (MPEG 400g/mol, CPEG 24% m/m, CCITRATO 20% m/m, pH 6) apresentou os melhores resultados K (9,24), Y (222,53%) e FP (1,93). A utilização do SDFA se mostrou eficiente para extração e purificação parcial da protease obtida por Aspergillus heteromorphus URM 0269 apresentando elevadas recuperações, indicando a possibilidade de aplicação na extração de proteases com potencial de aplicação industrial, especialmente na indústria de alimentos.

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Aqueous two‐phase systems in Latin America: perspective and future trends

Picó

Lima

Mussagy

et al. 2021

J of Chemical Tech & Biotech

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Aqueous two‐phase systems (ATPS) have been demonstrated to be a simple and robust worldwide applied bioprocessing technology. From high value microscopic to macroscopic biological molecules obtained from a wide broad of matrices, ATPS have shown a high efficiency capacity for their concentration, enrichment, recovery, and purification. Particularly in Latin America, four relevant countries have paved the way and played important leading roles in this transformation history. This perspective article aims to present a critical analysis of the practical evolution of the main implementations of ATPS in Argentina, Brazil, Chile, and Mexico. Special attention is given to novel applications of this biphasic bioprocess that are being explored to accomplish visionary therapeutic tasks for the healthcare, biotechnology, and pharmaceutical sectors, as well as other environmental purposes. © 2021 Society of Chemical Industry (SCI).

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Purification and characterization of a novel Aspergillus heteromorphus URM 0269 protease extracted by aqueous two-phase systems PEG/citrate

Cited by 16 publications

References 40 publications

Use of a Sequential Fermentation Method for the Production of Aspergillus tamarii URM4634 Protease and a Kinetic/Thermodynamic Study of the Enzyme

Use of a Sequential Fermentation Method for the Production of Aspergillus tamarii URM4634 Protease and a Kinetic/Thermodynamic Study of the Enzyme

PURIFICAÇÃO DE PROTEASES DE Aspergillus heteromorphus URM 0269 POR SISTEMA DE DUAS FASES AQUOSAS PEG-Citrato

Aqueous two‐phase systems in Latin America: perspective and future trends

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