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Xylan is the hemicellulose with the highest incidence in agricultural and agroindustrial residues. Xylooligosaccharides (XOS) are xylose oligomers recognized for their prebiotic potential, in addition to other biological properties that promote health. XOS were produced through enzymatic hydrolysis and their prebiotic effect was evaluated. Mostly, XOS with a high degree of polymerization were produced. The microorganisms used were able to grow in the medium containing the XOS produced.

Made available in DSpace on 2021-06-25T10:56:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Xylooligosaccharides (XOS) are xylose oligomers, recognized for their prebiotic effect, with the ability to impact health by influencing the gut microbiota. This study aimed to produce XOS by xylan enzymatic hydrolysis and evaluate the prebiotic effect. Endoxylanase was produced by Aspergillus versicolor and purified, and the xylan from banana pseudostem was alkali extracted. XOS production was performed through enzymatic hydrolysis by varying the parameters of xylanase load, substrate mass load, and reaction time. Lactobacillus plantarum and Lactobacillus fermentum were grown using XOS as a carbon source. Using an enzymatic load of 30 IU. g−1, it was possible to obtain a good amount of XOS and good yield, around 11 g.L−1, and 61%, respectively. Regarding the substrate concentration, the increase of concentration also increased XOS production, however, decreased the yield. There was not significant XOS production until 16 h and production was similar after 24 h. XOS with a higher degree of polymerization were mainly produced. Both probiotic bacteria were able to grow on XOS containing medium. Up to 48 h, the growth was similar, of 1010 CFU/mL, with glucose and XOS as the carbon source. The growth with XOS increased after 48 h, while the growth with glucose remained stationary. This study demonstrates that XOS from banana pseudostem xylan presents prebiotic properties by increasing probiotic bacteria growth. Biosciences Institute Biochemistry and Microbiology Department São Paulo State University (UNESP) Institute for Research in Bioenergy (IPBEN) São Paulo State University (UNESP), R. 10, 2527, Santana School of Pharmaceutical Sciences (FCF) Department of Bioprocesses Engineering and Biotechnology São Paulo State University (UNESP) Biosciences Institute Biochemistry and Microbiology Department São Paulo State University (UNESP) Institute for Research in Bioenergy (IPBEN) São Paulo State University (UNESP), R. 10, 2527, Santana School of Pharmaceutical Sciences (FCF) Department of Bioprocesses Engineering and Biotechnology São Paulo State University (UNESP) CAPES: 001 FAPESP: 2017/22401-8 FAPESP: 2019/12997-6

In this study, Penicillium sp. section Gracilenta CBMAI 1583 was used to produce lipase under submerged conditions. The enzyme was purified and the biochemical properties of both the crude and purified enzymes were evaluated. Maximum lipase production (1.62 U mL−1) was obtained using olive oil 0.5% (w v−1) after 72 h of cultivation, representing a 90% increase in the lipase initially produced. The enzyme was purified using hydrophobic interaction chromatography (phenyl Sepharose) under conditions which allowed its interfacial activation. The partially purified sample showed an enzyme with esterase activity (65.4 kDa) on α- and β-naphthyl acetate and other with lipase activity (52.9 kDa) on octyl oleate. Optimum activity of crude and purified lipase was observed at pH 4.0 and 70 °C. The purified lipase was activated by NaCl, BaCl2, NH4Cl, MnSO4 and MgSO4; it also presented high stability in organic solvents such as hexane, 2.2.4-trimethylpentane, acetone, DMSO and toluene. Maximum enzyme activity was observed with p-nitrophenyl decanoate as substrate; and the enzyme kinetics showed to be directly affected by Triton X-100. The enzyme shows potential application in processes that operate in acid pH, such as treatment of dairy and industry effluents, resolution of esters in the pharmaceutical industry or in the food industry, as well in synthesis reaction under non-aqueous conditions., The authors gratefully acknowledge to São Paulo Research Foundation - FAPESP, Brazil, for the scholarship granted to the first author and Spanish Ministry of Science and Innovation - MICINN, Spain (Project BIO-2012–36861).

Made available in DSpace on 2018-12-11T17:15:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-02-01 Microbial lipases are important enzymes in food and pharmaceutical industries. In this work, a Candida viswanathii lipase was purified by hydrophobic interaction chromatography on octyl Sepharose. The purification presented 78.4% yield and the enzyme was 8.7-fold purified, with specific activity 700.4 U/mg protein and 69 kDa molecular weight. Immobilization of the enzyme on the same support presented 72.5% yield and a derivative with 101% expressed activity (109.2 U/g support), indicating hyperactivation of the enzyme. Optimal activity for both free and immobilized lipase was observed at pH 4.0 and 45 °C. The free and immobilized lipase showed broad-range stability from acid to neutral pH, and apparent activation and stability on organic solvents. The derivative was 60-fold thermostabilized in relation to the free enzyme and fully retained its activity after four cycles of p-nitrophenyl palmitate hydrolysis. Slight activation was observed with dithiothreitol and β-mercaptoethanol. The free and immobilized lipase efficiently hydrolyzed monoesters, simple and mixed long chain triacylglycerols, as well as soy lecithin. The activity and stability in acid pH, the organic solvent tolerance and the lecithin hydrolysis indicate high potential application of the enzyme and its derivative in textile, food and pharma industries and for chemical synthesis. Bioprocess Engineering and Biotechnology Federal University of Tocantins – UFT, Rua Badejós, Chácaras 69/72, Zona Rural Biochemistry and Microbiology Department Bioscience Institute São Paulo State University IB/UNESP, Av. 24-A, 1515, Bela Vista Environmental Studies Center São Paulo State University CEA/UNESP, Av. 24-A, 1515, Bela Vista Biochemistry and Microbiology Department Bioscience Institute São Paulo State University IB/UNESP, Av. 24-A, 1515, Bela Vista Environmental Studies Center São Paulo State University CEA/UNESP, Av. 24-A, 1515, Bela Vista

Made available in DSpace on 2020-12-12T01:50:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Hemicelluloses are mainly branched heteropolysaccharides composed by xylose, arabinose, mannose, galactose, rhamnose and glucose. The main hemicellulose is xylan, a structure composed by xylose main chain branched with other monosaccharides or glucuronic and galacturonic acids. Among various hydrolytic enzymes acting on the degradation of xylan, α-L-arabinofuranosidases catalyze the hydrolysis of linkages between α-L-arabinofuranosides and the polysaccharide main chain. In this study, Aspergillus hortai strain CRM1919, isolated from soil surrounding saline and alkaline lagoons of the Nhecolândia region in Pantanal Matogrossense was investigated regarding the production of α-L-arabinofuranosidases. A. hortai produced α-L-arabinofuranosidase at high levels when cultivated in medium containing citrus pulp and orange peel at 1% (w/v) under static submerged cultures for 4 days and at pH 2.5 and 30 °C. The optimization steps increased 15-fold the enzymatic activity. The purified α-L-arabinofuranosidase was optimally active at pH of 4.0 and 60 °C, presenting half-lives of 265 and 230 min at 30 and 40 °C, respectively. High enzyme stability was observed after 24 h incubation at pH 5.0 and in presence of high concentration of NaCl in the reaction medium. The purified enzyme was mainly active against ρ-nitrophenyl-α-L-arabinofuranoside presenting the following kinetic parameters Km of 8.73 mmol/L, Vmax of 7.91 μmol/min.mg of protein and Kcat of 0.59/min. Hydrolysis of oat xylan performed in the presence of the purified α-arabinofuranosidase individually and in association with a purified xylanase made possible to verify the cooperation between these xylanolytic enzymes and the sequential action of each enzyme in the xylan structure hydrolysis. Bioenergy Research Institute Univ Estadual Paulista - UNESP, São Paulo Biochemistry and Microbiology Department Bioscience Institute Univ Estadual Paulista - UNESP, São Paulo Department of Biochemistry and Tissue Biology Institute of Biology Univ Campinas – UNICAMP, São Paulo Bioenergy Research Institute Univ Estadual Paulista - UNESP, São Paulo Biochemistry and Microbiology Department Bioscience Institute Univ Estadual Paulista - UNESP, São Paulo