Optimization of ß-Fructofuranosidase Production from Agrowaste by Aspergillus carbonarius and Its Application in the Production of Inverted Sugar.

RESEARCH BACKGROUND: Microbial ß-fructofuranosidases are widely employed in food industry to produce inverted sugar or fructooligosaccharides. In this study, a newly isolated Aspergillus carbonarius PC-4 strain was used to optimize the ß-fructofuranosidase production in a cost-effective process and the sucrose hydrolysis was evaluated to produce inverted sugars. EXPERIMENTAL APPROACH: Optimization of nutritional components of culture medium was carried out using simplex lattice mixture design for 72 and 120 h at 28 °C. One-factor-at-a-time methodology was used to optimize the physicochemical parameters. Crude enzyme was used for sucrose hydrolysis at different concentrations. RESULTS AND CONCLUSIONS: The optimized conditions of enzyme production were achieved from cultivations containing pineapple crown waste (1.3%, m/V) and yeast extract (0.3%, m/V) after 72 h with an enzyme activity of 9.4 U/mL, obtaining R2=91.85%, R2 adjusted=85.06%, highest F-value (13.52) and low p-value (0.003). One-factor-at-a-time used for optimizing the physicochemical conditions showed optimum temperature (20 °C), pH (5.5), agitation (180 rpm) and time course (72 h) with a 3-fold increase of enzyme production. The invertase-induced sucrose hydrolysis showed the maximum yield (3.45 mmol of reducing sugars) using 10% of initial sucrose concentration. Higher sucrose concentrations caused the inhibition of invertase activity, possibly due to the saturation of substrate or formation of sucrose aggregates, making it difficult for the enzyme to access sucrose molecules within the created clusters. Therefore, a cost-effective method was developed for the invertase production using agroindustrial waste and the produced enzyme can be used efficiently for inverted sugar production at high sucrose concentration. NOVELTY AND SCIENTIFIC CONTRIBUTION: This study presents an efficient utilization of pineapple crown waste to produce invertase by a newly isolated Aspergillus carbonarius PC-4 strain. This enzyme exhibited a good potential for inverted sugar production at high initial sucrose concentration, which is interesting for industrial applications.

ß-Fructofuranosidase and ß -D-Fructosyltransferase from New Aspergillus carbonarius PC-4 Strain Isolated from Canned Peach Syrup: Effect of Carbon and Nitrogen Sources on Enzyme Production.

ß-fructofuranosidase (invertase) and ß-D-fructosyltransferase (FTase) are enzymes used in industrial processes to hydrolyze sucrose aiming to produce inverted sugar syrup or fructooligosaccharides. In this work, a black Aspergillus sp. PC-4 was selected among six filamentous fungi isolated from canned peach syrup which were initially screened for invertase production. Cultivations with pure carbon sources showed that invertase and FTase were produced from glucose and sucrose, but high levels were also obtained from raffinose and inulin. Pineapple crown was the best complex carbon source for invertase (6.71 U/mL after 3 days of cultivation) and FTase production (14.60 U/mL after 5 days of cultivation). Yeast extract and ammonium chloride nitrogen sources provided higher production of invertase (6.80 U/mL and 6.30 U/mL, respectively), whereas ammonium nitrate and soybean protein were the best nitrogen sources for FTase production (24.00 U/mL and 24.90 U/mL, respectively). Fermentation parameters for invertase using yeast extract were Y P/S = 536.85 U/g and P P = 1.49 U/g/h. FTase production showed values of Y P/S = 2,627.93 U/g and P P = 4.4 U/h using soybean protein. The screening for best culture conditions showed an increase of invertase production values by 5.10-fold after 96 h cultivation compared to initial experiments (fungi bioprospection), while FTase production increased by 14.60-fold (44.40 U/mL) after 168 h cultivation. A. carbonarius PC-4 is a new promising strain for invertase and FTase production from low cost carbon sources, whose synthesized enzymes are suitable for the production of inverted sugar, fructose syrups, and fructooligosaccharides.

Evaluation of the fungitoxic activity of vegetal extracts on the mycelial growth of phytopathogens / Avaliação da atividade fungitóxica de extratos vegetais sobre o crescimento micelial de fitopatógenos

Although the productivity of common bean in Tocantins is economically favorable, it has been infected by various pathogens found in soil. Among the major diseases is the web blight and root rot caused by Rhizoctonia solani and collar rot caused by the fungus Sclerotium rolfsii. This study aimed to evaluate the fungitoxic activity of methanol extracts of eight plant species on the inhibition of mycelial growth of S. rolfsii and R. solani. The fungitoxic activities were carried out over the inhibition of mycelial growth by means in vitro assays. The extracts were applied in concentrations of 250, 500, 1000, 2500 e 5000 µg ml-1 in PDA culture medium. In bioassays, it was found the significant effect of plant, concentration and also their interaction on the antifungal activity of the extracts. However, some extracts showed no inhibition of mycelial growth of the pathogens studied. Among those who had higher inhibitions is the extract of Lantana trifolia, which inhibited the mycelial growth of S. rolfsii in all concentrations, being the same as 97% for the highest concentration. When the methanol extract of Piper amplum Kunth, inhibition of the highest concentration was 83% for S. rolfsii and 74% for R. solani. These results show the potential of methanolic extract of Lantana trifolia and Piper amplum Kunth in the control set of plant pathogens studied.

Apesar da produtividade do feijão comum no Tocantins ser economicamente favorável, o mesmo pode ser infectado por vários patógenos habitantes do solo, dentre as principais doenças encontra-se a mela e a podridão radicular causadas pelo fungo Rhizoctonia solani e a podridão do colo causada pelo fungo Sclerotium rolfsii. Este trabalho teve como objetivo avaliar a atividade fungitóxica dos extratos metanólicos de oito espécies vegetais sobre a inibição do crescimento micelial de Sclerotium rolfsii e Rhizoctonia solani. As atividades fungitóxicas foram realizadas perante a inibição do crescimento micelial por meio de ensaios in vitro, sendo os extratos aplicados nas concentrações de 250, 500, 1000, 2500 e 5000 µg ml-1 em meio de cultura BDA. Observou-se o efeito significativo dos fatores planta, concentração e também da interação destes sobre as atividades fungitóxicas. No entanto, alguns extratos não apresentaram inibição do crescimento micelial dos fitopatógenos estudados. Entre os que apresentaram maiores inibições encontra-se o extrato de Lantana trifolia, que inibiu o crescimento micelial do S. rolfsii em todas as concentrações, sendo o mesmo de 97% para a maior concentração. Já a concentração mais elevada do extrato metanólico de Piper amplum apresentou inibição de 83% sobre o crescimento micelial de S. rolfsii e 74% sobre o crescimento micelial de R. solani. Tais resultados evidenciam a potencialidade dos extratos metanólicos das folhas de Lantana trifolia e de Piper amplum no controle dos fitopatógenos estudados.