White rot fungi have one of the most efficient oxidative lignin degradation systems, and present an enzymatic complex that is responsible for digesting lignocellulosic matter. The aim of this study was to evaluate the production of laccase and total peroxidases by Ganoderma lucidum via solid-state fermentation using açaí seeds and marupá (Simarouba amara) sawdust unsupplemented and supplemented with wheat, corn and rice bran. G. lucidum was inoculated in substrates prepared with the residues and incubated at 25 ºC. Enzymatic extractions were performed on the culture substrates every 2 days for 30 days. The enzymatic activities were determined using ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt), with the addition of the enzymatic cofactor H2O2 for total peroxidases. The laccase activity was higher in the supplemented residues, with emphasis on the açaí-based substrate on the 16th day of cultivation, while in marupá the maximum activity was on the 6th day. In the unsupplemented açaí residue, the maximum peak of activity was on the 8th day and, in marupá, no fungal growth was observed. As for total peroxidases, G. lucidum cultivated in the supplemented açaí substrates showed activity peaks on the 8th, 12th, 16th and 28th day, and on 6th and 12th day under unsupplemented conditions. While, in the marupá, total peroxidase activity was observed only on the 6th day of cultivation. Thus, G. lucidum showed potential for producing laccases and total peroxidases, with the substrate supplementation inducing the synthesis of these enzymes.
Ganoderma lucidum is a medicinal mushroom widely recognized as a source of biomolecules with pharmacological properties, however, little is known about the factors that influence the synthesis of bioactive proteins by this fungus when cultivated under submerged fermentation. The objective of this work was to evaluate the production of mycelial biomass and intracellular proteases and protease inhibitors by G. lucidum cultivated under different submerged fermentation conditions. The cultivation was carried out in a medium composed of glucose (10 or 20 g.L-1), soy peptone (2.5 or 5 g.L-1) and yeast extract (5 g.L-1), with incubation under agitation (120 rpm) and non-agitation, totaling 8 experimental conditions. Biomass production was determined from the dry weight, while glucose consumption was estimated by quantification of reducing sugars. The proteins were extracted in NaCl (0.15 M), and the protein extracts were submitted to protein quantification by the Bradford method, total proteolytic activity using azocasein, caseinolytic and fibrinolytic activity in Petri dishes, activity of serine (trypsin and chymotrypsin) and cysteine (papain) protease inhibitors. Cultivation in agitated condition showed higher biomass production with a maximum value of 7 g.L-1, in addition to higher activities of trypsin, chymotrypsin and papain inhibitors, with 154 IU.mg-1, 153 IU.mg-1 e 343 IU.mg-1 of protein, respectively. The non-agitated condition showed a greater potential for obtaining proteins, total proteases, caseinolytic and fibrinolytic enzymes, with maximum values of 433 mg.g-1 of extract, 71 U.mL-1 of extract, 63.62 mm2 and 50.27 mm2, respectively. Thus, a medium composed of soy peptone, yest extract and glucose in a 1:2:4 proportion is recommended, under agitation to produce protease inhibitors, and the non-agitated condition when the target is, mainly caseinolytic and fibrinolytic enzymes.
The choice of residue to be used in the cultivation of a fungal species must take into account the structural and chemical composition of the material in order to be successful in their cultivation. Thus, this study evaluated the physical and chemical characteristics of pineapple and açaí residues and their alterations after the cultivation of two Pleurotus ostreatus strains (474 and 542). The fungi were cultivated in pineapple, açaí, and pineapple + açaí residues, which were added to a mixture of brans, and CaCO3 in the proportion of 78:20:2 w/w/w. The physical and chemical parameters and proximate composition of the residues, and the initial and spent substrates were determined, in addition to being analyzed using scanning electron microscopy and X-ray diffraction. In general, among the residues tested, the pineapple substrate showed more interesting nutritional characteristics for fungiculture, as well as structural conformation and greater availability of nutrients. The açaí substrate was not suitable for fungal cultivation due to its structural characteristics and high tannin content. In this sense, knowing the characteristics of the material used as a support for mushroom growth is of paramount importance in order to achieve successful cultivation.
Este artigo apresenta uma abordagem sobre o uso de controle biológico aplicando a ação de bactérias contra Corynespora cassiicola (Bert & Curt) Wei pois este é um dos principais fitopatógenos de propagação aérea que limita a produtividade das solanáceas. A utilização de bactérias no controle de doenças em solanáceas é uma metodologia em difusão, pois o biocontrole de fitopatógenos realizado por bactérias pode ocorrer por diferentes mecanismos como a inibição por antibióticos, o parasitismo, a competição pelo espaço físico ou a indução de resistência da planta ao ataque de patógenos. Esta pesquisa visou selecionar, identificar e avaliar in vitro e in planta bactérias antagonistas com potencial para desenvolvimento de um bioproduto que contribua para o desenvolvimento agrícola sustentável de tomate. As coletas e os testes in planta foram realizadas na Estação Experimental de Hortaliças Dr. Alejo von Der Pahlen, KM 14 da estrada AM 010 (S: 02° 59’ 45,7” e W: 60° 01’ 22,3”). Os testes in vitro foram realizados na Embrapa Amazônia Ocidental. Os resultados indicam que o Bacillus amyloliquefaciens, apresentou 75,73% de inibição do C. cassiicola in vitro e 56% de controle da mancha-alvo quando comparado ao Microbacterium resistens e a Stenotrophomonas maltophilia nos testes em tomateiro.
Ganoderma lucidum is a basidiomycete whose production is of great interest due to its medicinal properties, and analyzing the potential of Amazonian lignocellulosic residues in the cultivation of G. lucidum is a way to enable the use of this material in mushroom cultivation and the production of biomolecules, including enzymes of commercial interest. The objective of this study was to evaluate the activity of cellulases and xylanases produced by G. lucidum when cultivated in açaí seed (Euterpe sp.) residues and marupá (Simarouba amara) sawdust under supplemented and non-supplemented conditions. Solid-state cultivation was carried out in 250 mL flasks containing 50 g of lignocellulosic residues, plus 2% of CaCO3, under supplemented (18% of rice, wheat and corn bran) and non-supplemented conditions. The flasks were incubated in a BOD incubator at 25 ºC. Enzyme extraction was performed from the fungus growth substrate over 30 days of cultivation, with collections every two days. The enzymatic extracts had their endoglucanase (CMCase), total cellulase (FPase) and xylanase activities determined. The highest enzymatic activity of CMCase and xylanase were 5.97 U/g and 1.90 U/g, respectively, on the 26th day of cultivation in the supplemented marupá sawdust substrate. While the maximum FPase content was 0.24 U/g, which was observed in the 8th day for the supplemented açaí substrate. Thus, the tested residues proved to be promising for the mycelial development of G. lucidum, with the supplementation with corn, rice and wheat bran being favorable to the production of enzymes with biotechnological interest.
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