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URI permanente desta comunidadehttps://thoth.dti.ufv.br/handle/123456789/3352
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Item Purification and characterization of a protease from Aspergillus sydowii URM5774: Coffee ground residue for protease production by solid state fermentation(Academia Brasileira de Ciências, 2021) Rocha, Felype T. B.; Brandão-Costa, Romero M. P.; Neves, Anna Gabrielly D.; Cardoso, Kethylen B. B.; Nascimento, Thiago P.; Albuquerque, Wendell W. C.; Porto, Ana Lúcia F.Solid state fermentation is a promising technology largely used in biotechnology process and is a suitable strategy for producing low-cost enzymatic products. At the present study, a novel enzyme obtained through solid state fermentation using Aspergillus sydowii was herein purifi ed and characterized. The fermentations used coffee ground residue as substrate and the crude enzyme was submitted through further purifi cation steps of: acetonic precipitation, DEAE-Sephadex and Superdex G-75 column. Both crude and purifi ed enzymes were submitted to biochemical characterization of their thermostability, optimal temperature and pH, effects of inhibitors and metal ions. A purifi ed protease was obtained with yield of 5.9-fold and 53% recovery, with maximal proteolytic activity of 352.0 U/mL. SDS-PAGE revealed a band of protein at 47.0 kDa. The enzyme activity was abolished in the presence of phenyl-methyl sulfonyl fl uoride and partially inhibited against Triton X-100 (78.0%). The optimal activity was found in pH 8.0 at 45°C of temperature. Besides, the enzyme showed stability between 35°C and 50°C. It was possible to determine appropriate conditions to the obtainment of thermostable proteases with biotechnological interest associated with a method that concomitantly shows excellent production levels and recovery waste raw material in a very profi table process.Item Large-scale, high-efficiency production of coffee somatic embryos(Crop Breeding and Applied Biotechnology, 2016-03) Maciel, Anna Lygia de Rezende; Rodrigues, Filipe Almendagna; Pasqual, Moacir; Carvalho, Carlos Henrique Siqueira deThe study aims to compare the efficiency of previously used liquid media for coffee, evaluating NAA concentrations in liquid medium and proline in semi-solid medium in the regeneration of somatic embryos and asses concentrations of BA and IAA in the maturation of embryos in temporary immer- sion bioreactors. For the regeneration of globular embryos from embryogenic aggregates and calli, we tested five concentrations (0.00, 0.25, 0.50, 1.00 and 2.00 mg L -1 ) of NAA in liquid medium and five concentrations (0.0, 0.5, 1.0, 2.0 and 4.0 g L -1 ) of proline in semisolid medium. The multiplication of embryogenic aggregates was highest in culture medium MM, reaching a density 7.5 times greater than that of the initial density. NAA promoted a linear increase in embryo regeneration. The medium containing 2.0 mg L -1 BA and 0.0 mg L -1 IAA yielded the highest percentage of large cotyledonary embryos.