Production of milk-coagulating protease by fungus Pleurotus djamor through solid state fermentation using wheat bran as the low-cost substrate.

Proteases are enzymes that hydrolyze peptide bonds present in proteins and peptides. They are widely used for various industrial applications, such as in the detergent, food, and dairy industries. Cheese is one of the most important products of the dairy industry, and the coagulation stage is crucial during the cheese-making process. Enzymatic coagulation is the most common technique utilized for this purpose. Microbial enzymes are frequently used for coagulation due to their advantages in terms of availability, sustainability, quality control, product variety, and compliance with dietary and cultural/religious requirements. In the present study, we identified and subsequently characterized milk coagulant activity from the fungus Pleurotus djamor PLO13, obtained during a solid-state fermentation process, using the agro-industrial residue, wheat bran, as the fermentation medium. Maximum enzyme production and caseinolytic activity occurred 120 h after cultivation. When the enzyme activity against various protease-specific synthetic substrates and inhibitors was analyzed, the enzyme was found to be a serine protease, similar to elastase 2. This elastase-2-like serine protease was able to coagulate pasteurized whole and reconstituted skim milk highly efficiently in the presence and absence of calcium, even at room temperature. The coagulation process was influenced by factors such as temperature, time, and calcium concentration. We demonstrate here, for the first time, an elastase-2-like enzyme in a microorganism and its potential application in the food industry for cheese production.

Use of waste frying oil and coconut pulp for the production, isolation, and characterization of a new lipase from Moesziomyces aphidis.

Lipases comprise the third most commercialized group of enzymes worldwide and those of microbial origin are sought for their multiple advantages. Agro-industrial waste can be an alternative culture medium for producing lipases, reducing production costs and the improper disposal of waste frying oil (WFO). This study aimed to produce yeast lipases through submerged fermentation (SF) using domestic edible oil waste as inducer and alternative culture medium. The optimal culture conditions, most effective inducer, and purification method for a new lipase from Moesziomyces aphidis BRT57 were identified. Yeast was cultured in medium containing green coconut pulp and WFO waste for 72 h. The maximum production of lipases in SF occurred in a culture medium containing WFO and yeast extract at 48 and 72 h of incubation, with enzyme activities of 8.88 and 11.39 U mL-1, respectively. The lipase was isolated through ultrafiltration followed by size exclusion chromatography, achieving a 50.46 % recovery rate. To the best of our knowledge, this is the first study to report the production and purification of lipases from M. aphidis, demonstrating the value of frying oil as inducer and alternative medium for SF, contributing to the production of fatty acids for biodiesel from food waste.

Aproveitamento da semente da jaca para a obtenção de endoglucanase a partir de Aspergillus níger por fermentação em estado sólido / The use of jackfruit seed for obtainment of endoglucanases from Aspergillus níger for fermentation in solid state

A jaca é alimento consumido por brasileiros sob diferentes formas: in natura, doces, conservas, dentre outros. Os resíduos derivados (sementes) desses produtos são ricos em nutrientes tais como proteínas, fibras, sais minerais e ácidos graxos podendo ser utilizados em diferentes alimentos como componentes enriquecedores, minimizando a poluição ambiental. O presente trabalho teve como objetivo avaliar o efeito da variação do teor de água (40%, 50% e 60%), do tempo de fermentação (24h, 48h, 72h e 96h) e da temperatura (25 °C, 30 °C e 35 °C), na produção de endoglucanase do fungo Aspergillus niger por fermentação em estado sólido, utilizando semente de jaca como substrato. As fermentações ocorreram em estufa bacteriológica, a endoglucanase foi quantificada como carboximetilcelulase utilizando CMC como substrato e o teor de proteína solúvel foi determinado pelo método de Bradford. O melhor resultado, atividade específica de 138,45 (U/mg), foi obtido nas condições de fermentação realizada com 60% do teor de água a 35 °C no tempo de 72h. A partir dos resultados obtidos foi possível concluir que a temperatura de fermentação foi o principal fator que afetou a produção da CMCase pelo Aspergillus niger a partir do semente de jaca.

Jackfruit is a kind of food eaten by many Brazilians in different ways: fresh, sweets, conserves, among others. The residue (seeds) derived from these products is rich in nutrients such as proteins, fibers, mineral salts and fatty acids, and it can be used in different kinds of food as enriching compounds, minimizing environmental pollution. The present study aimed to evaluate the effect of water percentage variation (40, 50, and 60%), time of fermentation (24, 48, 72, and 96 hours), and temperature (25, 30, and 35ºC) in endoglucanases production for Aspergillus niger fungus, through fermentation in solid state, having jackfruit seed as substratum. The fermentations took place in bacteriological greenhouse; endoglucanases was quantified as carboximetil-cellulose using CMC as substratum, and the percentage of solvable protein was determined by Bradford method. The best result, specific activity 138,45 (U/mg), was obtained at the conditions of fermentation realized with 60% of water percentage, at 35ºC, in 72 hours. Departing from results, it was possible to conclude that the temperature of fermentation was the main coefficient to affect the production of CMCase by Aspergillus niger from jackfruit seed.