Aspergillus phoenicis é um fungo termotolerante interessante, uma vez que pode sintetizar enzimas com diversas aplicações em indústrias alimentícias em função de seu grande potencial de hidrólise. Neste trabalho, verificou-se que esse fungo produziu níveis enzimáticos elevados, quando o mesmo foi cultivado em meio de cultura de baixo custo, constituído de flocos de diferentes origens, como farinha de mandioca, fibra de trigo, soja triturada, resíduos agroindustriais, amido e glicose ou maltose. Diversos sistemas enzimáticos foram produzidos a partir dessas fontes de carbono, mas a amilase foi a mais evidente, seguida de pectinase e xilanase. Traços de atividades de CMcases, avicelase, lipase, β-xilosidase, β-glicosidase e α-glicosidase também foram detectados. Amilases foram produzidas em flocos de centeio, amido, flocos de aveia, flocos de milho, farinha de mandioca e fibra de trigo. Níveis amilolíticos significantes foram produzidos em meio de cultura contendo glicose ou quando esse açúcar não estava mais presente, sugerindo uma forma constitutiva dessa enzima. Farinha de mandioca, flocos de centeio, aveia, cevada e milho também foram utilizados como substratos nas reações de hidrólise, com o objetivo de verificar a potencial liberação de açúcares redutores. Flocos de milho induziram maior formação de açúcares redutores, quando comparados com os outros substratos. A análise por cromatografia em camada delgada dos produtos finais da reação revelou que a hidrólise da farinha de mandioca liberou malto-oligossacarídeos, mas farinha de mandioca e flocos de milho, centeio, aveia e cevada foram hidrolisados até glicose. Esses resultados sugeriram a presença de glucoamilase e α-amilase como parte da mistura enzimática de A. phoenicis.

Aspergillus phoenicis is an interesting heat tolerant fungus that can synthesize enzymes with several applications in the food industry due to its great hydrolytic potential. In this work, the fungus produced high enzymatic levels when cultivated on inexpensive culture media consisting of flakes from different origins such as cassava flour, wheat fibre, crushed soybean, agro-industrial wastes, starch, glucose or maltose. Several enzymatic systems were produced from these carbon sources, but amylase was the most evident, followed by pectinase and xylanase. Traces of CMCases, avicelase, lipase, β-xylosidase, β-glucosidase and α-glucosidase activities were also detected. Amylases were produced on rye flakes, starch, oat flakes, corn flakes, cassava flour and wheat fibre. Significant amylolytic levels were produced in the culture medium with glucose or when this sugar was exhausted, suggesting an enzyme in the constitutive form. Cassava flour, rye, oats, barley and corn flakes were also used as substrates in the hydrolytic reactions, aiming to verify the liberation potential of reducing sugars. Corn flakes induced greater liberation of reducing sugars as compared to the others. Thin layer chromatography of the reaction end products showed that the hydrolysis of cassava flour liberated maltooligosaccharides, but cassava flour and corn, rye, oats and barley flakes were hydrolyzed to glucose. These results suggested the presence of glucoamylase and α-amylase as part of the enzymatic pool of A. phoencis.

The filamentous fungus A. phoenicis produced high levels of ?-D-fructofuranosidase (FFase) when grown for 72 hrs under Solid-State Fermentation (SSF), using soy bran moistened with tap water (1:0.5 w/v) as substrate/carbon source. Two isoforms (I and II) were obtained, and FFase II was purified 18-fold to apparent homogeneity with 14% recovery. The native molecular mass of the glycoprotein (12% of carbohydrate content) was 158.5 kDa with two subunits of 85 kDa estimated by SDS-PAGE. Optima of temperature and pH were 55�C and 4.5. The enzyme was stable for more than 1 hr at 50�C and was also stable in a pH range from 7.0 to 8.0. FFase II retained 80% of activity after storage at 4�C by 200 hrs. Dichroism analysis showed the presence of random and ?-sheet structure. A. phoenicis FFase II was activated by Mn2+, Mg2+ and Co2+, and inhibited by Cu2+, Hg2+ and EDTA. The enzyme hydrolyzed sucrose, inulin and raffinose. Kd and Vmax values were 18 mM and 189 U/mg protein using sucrose as substrate.

Fungi collected from Brazilian soil and decomposing plants were screened for pectinase production. R. microsporus var. rhizopodiformis was the best producer and was selected to evaluate the pectic enzyme production under several nutritional and environmental conditions. The pectinase production was studied at 40ºC, under 28 carbon sources-supplemented medium. The inducer effect of several agro-industrial residues such as sugar cane bagasse, wheat flour and corncob on polygalacturonase (PG) activity was 4-, 3- and 2-fold higher than the control (pectin). In glucose-medium, a constitutive pectin lyase (PL) activity was detected. The results demonstrated that R. microsporus produced high levels of PG (57.7 U/mg) and PL (88.6 U/mg) in lemon peel-medium. PG had optimum temperature at 65 ºC and was totally stable at 55 ºC for 90 min. Half-life at 70 ºC was 68 min. These results suggested that the versatility of waste carbon sources utilization by R. microsporus, produce pectic enzymes, which could be useful to reduce production costs and environmental impacts related to the waste disposal.

O fungo filamentoso Aspergillus caespitosus foi um bom produtor de invertases intracelular e extracelular em fermentação submersa (FSbm) ou em estado sólido (FES), usando resíduos agroindustriais como fonte de carbono, sendo que para ambas as condições de cultivo, a maior produtividade foi obtida empregandose farelo de trigo. A produção da forma extracelular em FES mantido a 30ºC, por 72 horas, foi aumentada usandose solução de sais SR (1:1, m/v) para umidificar o substrato, sendo aproximadamente 5,5 vezes maior se comparada a FSbm (Meio Khanna) com a mesma fonte de carbono. Entretanto, a mistura de farelo de trigo e farinha de aveia em FES levou a um aumento de 2,2 vezes na produção enzimática se comparada ao uso isolado do farelo de trigo. A produção enzimática, em ambas as condições de cultivo, foi afetada pela adição suplementar de fontes de nitrogênio e fosfato. A adição de glicose em FSbm e em FES promoveu a diminuição da enzima extracelular, mas favoreceu um acúmulo intracelular de 35 vezes maior. A temperatura ótima de atividade para as invertases produzidas em FES e em FSbm foi de 50ºC e 60ºC, respectivamente, sendo estáveis a 50ºC por mais de 60 minutos. Todas as formas enzimáticas apresentaram atividade máxima em uma faixa de pH de 4.0-6.0.

The filamentous fungus Aspergillus caespitosus was a good producer of intracellular and extracellular invertases under submerged (SbmF) or solid-state fermentation (SSF), using agroindustrial residues, such as wheat bran, as carbon source. The production of extracellular enzyme under SSF at 30ºC, for 72h, was enhanced using SR salt solution (1:1, w/v) to humidify the substrate. The extracellular activity under SSF using wheat bran was around 5.5-fold higher than that obtained in SbmF (Khanna medium) with the same carbon source. However, the production of enzyme with wheat bran plus oat meal was 2.2-fold higher than wheat bran isolated. The enzymatic production was affected by supplementation with nitrogen and phosphate sources. The addition of glucose in SbmF and SSF promoted the decreasing of extracellular activity, but the intracellular form obtained in SbmF was enhanced 3-5-fold. The invertase produced in SSF exhibited optimum temperature at 50ºC while the extraand intracellular enzymes produced in SbmF exhibited maximal activities at 60ºC. All enzymatic forms exhibited maximal activities at pH 4.0-6.0 and were stable up to 1 hour at 50ºC.