Microbial Glycosidases for Nondigestible Oligosaccharides Production

There is much interest in the study and production of nondigestible oligosaccharides (NDOs), due to their bioactivities and beneficial effects to the human health. The main approach in the production of NDOs relies on the action of glycosidases performing hydrolysis or transglycosylation of polysaccharides and sugars. In this chapter, a description of the main microbial glycosidases used for NDOs production, their sources, their principal properties, and a description of the production processes with the better results obtained are discussed

Selective synthesis of citrus flavonoids prunin and naringenin using heterogeneized biocatalyst on graphene oxide

[EN] Production of citrus flavonoids prunin and naringenin was performed selectively through the enzyme hydrolysis of naringin, a flavonoid glycoside abundant in grapefruit wastes. To produce the monoglycoside flavonoid, prunin, crude naringinase from Penicillium decumbens was purified by a single purification step resulting in an enzyme with high -rhamnosidase activity. Both crude and purified enzymes were covalently immobilized on graphene oxide. The activity of the immobilized enzymes at different pH levels and temperatures, and the thermal stability were determined and compared with those exhibited by the free naringinases using specific substrates: p-nitrophenyl--d-glucoside (Glc-pNP) and p-nitrophenyl-alpha-l-rhamnopyranoside (Rha-pNP). The crude and purified naringinase supported on GO were tested in the hydrolysis of naringin, giving naringenin and prunin, respectively, in excellent yields. The supported enzymes can be reused many times without loss of activity. The naringinase stabilized on GO has high potential to produce the valuable citrus flavonoids prunin and naringenin.Authors acknowledge the financial support from MICINN Project CTQ-2015-67592-P and Program Severo Ochoa (SEV-2016-0683). JVC thanks Universitat Politecnica de Valencia for predoctoral fellowships. JY and AC thank the support from the National Natural Science Foundation of China (Grant No. 21320102001) and the 111 Project (Grant No. B17020).Carceller-Carceller, JM.; Martínez Galán, JP.; Monti, R.; Bassan, JC.; Filice, M.; Iborra Chornet, S.; Yu, J.... (2019). Selective synthesis of citrus flavonoids prunin and naringenin using heterogeneized biocatalyst on graphene oxide. Green Chemistry. 21(4):839-849. https://doi.org/10.1039/c8gc03661fS839849214Puri, M., & Banerjee, U. C. (2000). Production, purification, and characterization of the debittering enzyme naringinase. Biotechnology Advances, 18(3), 207-217. doi:10.1016/s0734-9750(00)00034-3Vila-Real, H., Alfaia, A. J., Rosa, M. E., Calado, A. R., & Ribeiro, M. H. L. (2010). An innovative sol–gel naringinase bioencapsulation process for glycosides hydrolysis. Process Biochemistry, 45(6), 841-850. doi:10.1016/j.procbio.2010.02.004C. Grassin and P.Fauquembergue , in Industrial Enzymology , ed. S. West and T. Godfrey , Nature Publishing Group , New York , 2nd edn, 1996 , p. 225Tsen, H.-Y., & Tsai, S.-Y. (1988). Comparison of the kinetics and factors affecting the stabilities of chitin-immobilized naringinases from two fungal sources. Journal of Fermentation Technology, 66(2), 193-198. doi:10.1016/0385-6380(88)90047-7SOARES, N. F. F., & HOTCHKISS, J. H. (1998). Naringinase Immobilization in Packaging Films for Reducing Naringin Concentration in Grapefruit Juice. Journal of Food Science, 63(1), 61-65. doi:10.1111/j.1365-2621.1998.tb15676.xPuri, M., Kaur, H., & Kennedy, J. F. (2005). Covalent immobilization of naringinase for the transformation of a flavonoid. Journal of Chemical Technology & Biotechnology, 80(10), 1160-1165. doi:10.1002/jctb.1303Norouzian, D., Hosseinzadeh, A., Inanlou, D. N., & Moazami, N. (1999). World Journal of Microbiology and Biotechnology, 15(4), 501-502. doi:10.1023/a:1008980018481Nishita, M., Park, S.-Y., Nishio, T., Kamizaki, K., Wang, Z., Tamada, K., … Minami, Y. (2017). Ror2 signaling regulates Golgi structure and transport through IFT20 for tumor invasiveness. Scientific Reports, 7(1). doi:10.1038/s41598-016-0028-xZhang, Y., Wu, C., Guo, S., & Zhang, J. (2013). Interactions of graphene and graphene oxide with proteins and peptides. Nanotechnology Reviews, 2(1), 27-45. doi:10.1515/ntrev-2012-0078Mathesh, M., Luan, B., Akanbi, T. O., Weber, J. K., Liu, J., Barrow, C. J., … Yang, W. (2016). Opening Lids: Modulation of Lipase Immobilization by Graphene Oxides. ACS Catalysis, 6(7), 4760-4768. doi:10.1021/acscatal.6b00942Li, W., Wen, H., Shi, Q., & Zheng, G. (2016). Study on immobilization of (+) γ-lactamase using a new type of epoxy graphene oxide carrier. Process Biochemistry, 51(2), 270-276. doi:10.1016/j.procbio.2015.11.030Hong, S.-G., Kim, J. H., Kim, R. E., Kwon, S.-J., Kim, D. W., Jung, H.-T., … Kim, J. (2016). Immobilization of glucose oxidase on graphene oxide for highly sensitive biosensors. Biotechnology and Bioprocess Engineering, 21(4), 573-579. doi:10.1007/s12257-016-0373-4Liu, F., Piao, Y., Choi, K. S., & Seo, T. S. (2012). Fabrication of free-standing graphene composite films as electrochemical biosensors. Carbon, 50(1), 123-133. doi:10.1016/j.carbon.2011.07.061Wang, Z., Zhou, X., Zhang, J., Boey, F., & Zhang, H. (2009). Direct Electrochemical Reduction of Single-Layer Graphene Oxide and Subsequent Functionalization with Glucose Oxidase. The Journal of Physical Chemistry C, 113(32), 14071-14075. doi:10.1021/jp906348xSingh, R. K., Kumar, R., & Singh, D. P. (2016). Graphene oxide: strategies for synthesis, reduction and frontier applications. RSC Advances, 6(69), 64993-65011. doi:10.1039/c6ra07626bVila-Real, H., Alfaia, A. J., Bronze, M. R., Calado, A. R. T., & Ribeiro, M. H. L. (2011). Enzymatic Synthesis of the Flavone Glucosides, Prunin and Isoquercetin, and the Aglycones, Naringenin and Quercetin, with Selective α-L-Rhamnosidase and β-D-Glucosidase Activities of Naringinase. Enzyme Research, 2011, 1-11. doi:10.4061/2011/692618Mamma, D., Kalogeris, E., Hatzinikolaou, D. G., Lekanidou, A., Kekos, D., Macris, B. J., & Christakopoulos, P. (2004). Biochemical Characterization of the Multi-enzyme System Produced byPenicillium decumbensGrown on Rutin. Food Biotechnology, 18(1), 1-18. doi:10.1081/fbt-120030382Chang, H.-Y., Lee, Y.-B., Bae, H.-A., Huh, J.-Y., Nam, S.-H., Sohn, H.-S., … Lee, S.-B. (2011). Purification and characterisation of Aspergillus sojae naringinase: The production of prunin exhibiting markedly enhanced solubility with in vitro inhibition of HMG-CoA reductase. Food Chemistry, 124(1), 234-241. doi:10.1016/j.foodchem.2010.06.024Yadav, S., Yadava, S., & Yadav, K. D. S. (2013). Purification and characterization of α-l-rhamnosidase from Penicillium corylopholum MTCC-2011. Process Biochemistry, 48(9), 1348-1354. doi:10.1016/j.procbio.2013.05.001Zhu, Y., Jia, H., Xi, M., Xu, L., Wu, S., & Li, X. (2017). Purification and characterization of a naringinase from a newly isolated strain of Bacillus amyloliquefaciens 11568 suitable for the transformation of flavonoids. Food Chemistry, 214, 39-46. doi:10.1016/j.foodchem.2016.06.108Zhang, T., Yuan, W., Li, M., Miao, M., & Mu, W. (2018). Purification and characterization of an intracellular α-l-rhamnosidase from a newly isolated strain, Alternaria alternata SK37.001. Food Chemistry, 269, 63-69. doi:10.1016/j.foodchem.2018.06.134Vila-Real, H., Alfaia, A. J., Rosa, J. N., Gois, P. M. P., Rosa, M. E., Calado, A. R. T., & Ribeiro, M. H. (2011). α-Rhamnosidase and β-glucosidase expressed by naringinase immobilized on new ionic liquid sol–gel matrices: Activity and stability studies. Journal of Biotechnology, 152(4), 147-158. doi:10.1016/j.jbiotec.2010.08.005Smith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., … Klenk, D. C. (1985). Measurement of protein using bicinchoninic acid. Analytical Biochemistry, 150(1), 76-85. doi:10.1016/0003-2697(85)90442-7Erickson, H. P. (2009). Size and Shape of Protein Molecules at the Nanometer Level Determined by Sedimentation, Gel Filtration, and Electron Microscopy. Biological Procedures Online, 11(1), 32-51. doi:10.1007/s12575-009-9008-xZhang, J., Zhang, F., Yang, H., Huang, X., Liu, H., Zhang, J., & Guo, S. (2010). Graphene Oxide as a Matrix for Enzyme Immobilization. Langmuir, 26(9), 6083-6085. doi:10.1021/la904014zMarolewski, A. (1996). Fundamentals of Enzyme Kinetics. Revised Edition By Athel Cornish-Bowden. Portland Press, London. 1995. xiii + 343 pp. 17.5 cm × 24.5 cm. ISBN 1-85578-072-0. $29.00. Journal of Medicinal Chemistry, 39(4), 1010-1011. doi:10.1021/jm9508447Romero, C., Manjón, A., Bastida, J., & Iborra, J. (1985). A method for assaying the rhamnosidase activity of naringinase. Analytical Biochemistry, 149(2), 566-571. doi:10.1016/0003-2697(85)90614-1Fox, D. W., Savage, W. L., & Wender, S. H. (1953). Hydrolysis of Some Flavonoid Rhamnoglucosides to Flavonoid Glucosides. Journal of the American Chemical Society, 75(10), 2504-2505. doi:10.1021/ja01106a507Miller, G. L. (1959). Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar. Analytical Chemistry, 31(3), 426-428. doi:10.1021/ac60147a030LAEMMLI, U. K. (1970). Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 227(5259), 680-685. doi:10.1038/227680a0Heukeshoven, J., & Dernick, R. (1985). Simplified method for silver staining of proteins in polyacrylamide gels and the mechanism of silver staining. Electrophoresis, 6(3), 103-112. doi:10.1002/elps.1150060302Sheldon, R. A., & van Pelt, S. (2013). Enzyme immobilisation in biocatalysis: why, what and how. Chem. Soc. Rev., 42(15), 6223-6235. doi:10.1039/c3cs60075

Covalent Immobilization of Naringinase over Two-Dimensional 2D Zeolites and its Applications in a Continuous Process to Produce Citrus Flavonoids and for Debittering of Juices

This is the peer reviewed version of the following article: J. M. Carceller, J. P. Martínez Galán, R. Monti, J. C. Bassan, M. Filice, J. Yu, M. J. Climent, S. Iborra, A. Corma, ChemCatChem 2020, 12, 4502, which has been published in final form at https://doi.org/10.1002/cctc.202000320. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.[EN] The crude naringinase from Penicillium decumbens and a purified naringinase with high a-L-rhamnosidase activity could be covalently immobilized on two-dimensional zeolite ITQ-2 after surface modification with glutaraldehyde. The influence of pH and temp. on the enzyme activity (in free and immobilized forms) as well as the thermal stability were detd. using the specific substrate: p-nitrophenyl-alpha-L-rhamnopyranoside (Rha-pNP). The crude and purified naringinase supported on ITQ-2 were applied in the hydrolysis of naringin, giving the flavonoids naringenin and prunin resp. with a conversion >90% and excellent selectivity. The supported enzymes showed long term stability, being possible to perform up to 25 consecutive cycles without loss of activity, showing its high potential to produce the valuable citrus flavonoids prunin and naringenin. We have also succeeded in the application of the immobilized crude naringinase on ITQ-2 for debittering grapefruit juices in a continuous process that was maintained operating for 300 h, with excellent results.The authors acknowledge financial support from PGC2018-097277-B-100 (MCIU/AEI/FEDER,UE) project and Program Severo Ochoa (SEV-2016-0683). Jilin agreement 111 Project (Grant No. B17020). JMC thanks to Universitat Politecnica de Valencia for predoctoral fellowships.Carceller-Carceller, JM.; Martínez Galán, JP.; Monti, R.; Bassan, JC.; Filice, M.; Yu, J.; Climent Olmedo, MJ.... (2020). Covalent Immobilization of Naringinase over Two-Dimensional 2D Zeolites and its Applications in a Continuous Process to Produce Citrus Flavonoids and for Debittering of Juices. ChemCatChem. 12(18):4502-4511. https://doi.org/10.1002/cctc.202000320S450245111218Puri, M., & Banerjee, U. C. (2000). Production, purification, and characterization of the debittering enzyme naringinase. Biotechnology Advances, 18(3), 207-217. doi:10.1016/s0734-9750(00)00034-3Vila-Real, H., Alfaia, A. J., Rosa, M. E., Calado, A. R., & Ribeiro, M. H. L. (2010). An innovative sol–gel naringinase bioencapsulation process for glycosides hydrolysis. Process Biochemistry, 45(6), 841-850. doi:10.1016/j.procbio.2010.02.004RoitNer, M., Schalkhammer, T., & Pittner, F. (1984). Preparation of prunin with the help of immobilized naringinase pretreated with alkaline buffer. Applied Biochemistry and Biotechnology, 9(5-6), 483-488. doi:10.1007/bf02798402Ribeiro, I. A., Rocha, J., Sepodes, B., Mota-Filipe, H., & Ribeiro, M. H. (2008). Effect of naringin enzymatic hydrolysis towards naringenin on the anti-inflammatory activity of both compounds. Journal of Molecular Catalysis B: Enzymatic, 52-53, 13-18. doi:10.1016/j.molcatb.2007.10.011Puri, M., Marwaha, S. S., Kothari, R. M., & Kennedy, J. F. (1996). Biochemical Basis of Bitterness in Citrus Fruit Juices and Biotech Approaches for Debittering. Critical Reviews in Biotechnology, 16(2), 145-155. doi:10.3109/07388559609147419Barbosa, O., Ortiz, C., Berenguer-Murcia, Á., Torres, R., Rodrigues, R. C., & Fernandez-Lafuente, R. (2015). Strategies for the one-step immobilization–purification of enzymes as industrial biocatalysts. Biotechnology Advances, 33(5), 435-456. doi:10.1016/j.biotechadv.2015.03.006Garcia-Galan, C., Berenguer-Murcia, Á., Fernandez-Lafuente, R., & Rodrigues, R. C. (2011). Potential of Different Enzyme Immobilization Strategies to Improve Enzyme Performance. Advanced Synthesis & Catalysis, 353(16), 2885-2904. doi:10.1002/adsc.201100534ONO, M., TOSA, T., & CHIBATA, I. (1978). Preparation and properties of immobilized naringinase using tannin-aminohexyl cellulose. Agricultural and Biological Chemistry, 42(10), 1847-1853. doi:10.1271/bbb1961.42.1847Tsen, H.-Y., & Tsai, S.-Y. (1988). Comparison of the kinetics and factors affecting the stabilities of chitin-immobilized naringinases from two fungal sources. Journal of Fermentation Technology, 66(2), 193-198. doi:10.1016/0385-6380(88)90047-7SOARES, N. F. F., & HOTCHKISS, J. H. (1998). Naringinase Immobilization in Packaging Films for Reducing Naringin Concentration in Grapefruit Juice. Journal of Food Science, 63(1), 61-65. doi:10.1111/j.1365-2621.1998.tb15676.xPuri, M., Kaur, H., & Kennedy, J. F. (2005). Covalent immobilization of naringinase for the transformation of a flavonoid. Journal of Chemical Technology & Biotechnology, 80(10), 1160-1165. doi:10.1002/jctb.1303Lei, S., Xu, Y., Fan, G., Xiao, M., & Pan, S. (2011). Immobilization of naringinase on mesoporous molecular sieve MCM-41 and its application to debittering of white grapefruit. Applied Surface Science, 257(9), 4096-4099. doi:10.1016/j.apsusc.2010.12.003Luo, J., Li, Q., Sun, X., Tian, J., Fei, X., Shi, F., … Liu, X. (2019). The study of the characteristics and hydrolysis properties of naringinase immobilized by porous silica material. RSC Advances, 9(8), 4514-4520. doi:10.1039/c9ra00075eNunes, M. A. P., Vila-Real, H., Fernandes, P. C. B., & Ribeiro, M. H. L. (2009). Immobilization of Naringinase in PVA–Alginate Matrix Using an Innovative Technique. Applied Biochemistry and Biotechnology, 160(7), 2129-2147. doi:10.1007/s12010-009-8733-6Busto, M. D., Meza, V., Ortega, N., & Perez-Mateos, M. (2007). Immobilization of naringinase from Aspergillus niger CECT 2088 in poly(vinyl alcohol) cryogels for the debittering of juices. Food Chemistry, 104(3), 1177-1182. doi:10.1016/j.foodchem.2007.01.033Huang, W., Zhan, Y., Shi, X., Chen, J., Deng, H., & Du, Y. (2017). Controllable immobilization of naringinase on electrospun cellulose acetate nanofibers and their application to juice debittering. International Journal of Biological Macromolecules, 98, 630-636. doi:10.1016/j.ijbiomac.2017.02.018Gong, X., Xie, W., Wang, B., Gu, L., Wang, F., Ren, X., … Yang, L. (2017). Altered spontaneous calcium signaling of in situ chondrocytes in human osteoarthritic cartilage. Scientific Reports, 7(1). doi:10.1038/s41598-017-17172-wCarceller, J. M., Martínez Galán, J. P., Monti, R., Bassan, J. C., Filice, M., Iborra, S., … Corma, A. (2019). Selective synthesis of citrus flavonoids prunin and naringenin using heterogeneized biocatalyst on graphene oxide. Green Chemistry, 21(4), 839-849. doi:10.1039/c8gc03661fPuri, M., Marwaha, S. S., & Kothari, R. M. (1996). Studies on the applicability of alginate-entrapped naringiase for the debittering of kinnow juice. Enzyme and Microbial Technology, 18(4), 281-285. doi:10.1016/0141-0229(95)00100-xNorouzian, D., Hosseinzadeh, A., Inanlou, D. N., & Moazami, N. (1999). World Journal of Microbiology and Biotechnology, 15(4), 501-502. doi:10.1023/a:1008980018481Saallah, S., Naim, M. N., Lenggoro, I. W., Mokhtar, M. N., Abu Bakar, N. F., & Gen, M. (2016). Immobilisation of cyclodextrin glucanotransferase into polyvinyl alcohol (PVA) nanofibres via electrospinning. Biotechnology Reports, 10, 44-48. doi:10.1016/j.btre.2016.03.003Cipolatti, E. P., Valério, A., Henriques, R. O., Moritz, D. E., Ninow, J. L., Freire, D. M. G., … de Oliveira, D. (2016). Nanomaterials for biocatalyst immobilization – state of the art and future trends. RSC Advances, 6(106), 104675-104692. doi:10.1039/c6ra22047aCorma, A., Fornes, V., & Rey, F. (2002). Delaminated Zeolites: An Efficient Support for Enzymes. Advanced Materials, 14(1), 71-74. doi:10.1002/1521-4095(20020104)14:13.0.co;2-wGallego, E. M., Portilla, M. T., Paris, C., León-Escamilla, A., Boronat, M., Moliner, M., & Corma, A. (2017). «Ab initio» synthesis of zeolites for preestablished catalytic reactions. Science, 355(6329), 1051-1054. doi:10.1126/science.aal0121Margarit, V. J., Díaz-Rey, M. R., Navarro, M. T., Martínez, C., & Corma, A. (2018). Direct Synthesis of Nano-Ferrierite along the 10-Ring-Channel Direction Boosts Their Catalytic Behavior. Angewandte Chemie, 130(13), 3517-3521. doi:10.1002/ange.201711418Barbosa, O., Ortiz, C., Berenguer-Murcia, Á., Torres, R., Rodrigues, R. C., & Fernandez-Lafuente, R. (2014). Glutaraldehyde in bio-catalysts design: a useful crosslinker and a versatile tool in enzyme immobilization. RSC Adv., 4(4), 1583-1600. doi:10.1039/c3ra45991hSmith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., … Klenk, D. C. (1985). Measurement of protein using bicinchoninic acid. Analytical Biochemistry, 150(1), 76-85. doi:10.1016/0003-2697(85)90442-7Marolewski, A. (1996). Fundamentals of Enzyme Kinetics. Revised Edition By Athel Cornish-Bowden. Portland Press, London. 1995. xiii + 343 pp. 17.5 cm × 24.5 cm. ISBN 1-85578-072-0. $29.00. Journal of Medicinal Chemistry, 39(4), 1010-1011. doi:10.1021/jm9508447Miller, G. L. (1959). Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar. Analytical Chemistry, 31(3), 426-428. doi:10.1021/ac60147a030Cheong, M. W., Liu, S. Q., Zhou, W., Curran, P., & Yu, B. (2012). Chemical composition and sensory profile of pomelo (Citrus grandis (L.) Osbeck) juice. Food Chemistry, 135(4), 2505-2513. doi:10.1016/j.foodchem.2012.07.012Fox, D. W., Savage, W. L., & Wender, S. H. (1953). Hydrolysis of Some Flavonoid Rhamnoglucosides to Flavonoid Glucosides. Journal of the American Chemical Society, 75(10), 2504-2505. doi:10.1021/ja01106a507Corma, A., Fornes, V., Pergher, S. B., Maesen, T. L. M., & Buglass, J. G. (1998). Delaminated zeolite precursors as selective acidic catalysts. Nature, 396(6709), 353-356. doi:10.1038/24592Camblor, M. A., Corma, A., & Valencia, S. (1998). Characterization of nanocrystalline zeolite Beta. Microporous and Mesoporous Materials, 25(1-3), 59-74. doi:10.1016/s1387-1811(98)00172-3Beck, J. S., Vartuli, J. C., Roth, W. J., Leonowicz, M. E., Kresge, C. T., Schmitt, K. D., … Schlenker, J. L. (1992). A new family of mesoporous molecular sieves prepared with liquid crystal templates. Journal of the American Chemical Society, 114(27), 10834-10843. doi:10.1021/ja00053a02

Perfil, evolução e desfecho dos pacientes atendidos pelo serviço de atendimento móvel de urgência

Objective: to analyze the profile, the evolution and outcome of patients served by the SAMU 192. Method: cross-sectional, exploratory and descriptive study. The sample consisted of data from 600 adult patients, served in the year 2015 by the SAMU and referred to another health service. Descriptive analysis was performed to identify the population profile, the evolution and the intra-hospital outcome; correlation testing between response time in the pre-hospital service and length of stay; and multiple logistic regression between response time and outcome. It used p < 0.05 as the significance level. Results: there was a predominance of care for clinical occurrences, male gender and average age of 55.5 years. After pre-hospital care, 50.2% of the surveyed patients remained under observation and 34.8% required hospitalization. The pre-hospital response time was positively correlated with the length of stay. The main outcome was medical discharge, with 41% of them being referred for the continuity of treatment. Conclusion: the analysis showed a profile of care provided to patients with an average age of 55 years and suffering from clinical diseases that required referral to a health unit, remaining under hospital observation for up to 12 hours. These results are important for the support of care flow protocols in the RUE, in order to reduce the overload of tertiary services.Objetivo: analisar perfil, evolução e desfecho dos pacientes atendidos pelo SAMU 192. Método: estudo transversal, exploratório e descritivo. A amostra constou dados de 600 pacientes adultos, atendidos no ano de 2015 pelo SAMU e encaminhados para outro serviço de saúde. Realizou-se análise descritiva para identificar o perfil da população, a evolução e o desfecho intra-hospitalar; teste de correlação entre o tempo resposta no serviço pré-hospitalar e o tempo de internação; e regressão logística múltipla entre o tempo resposta e o desfecho. Utilizou-se p < 0,05 como nível de significância. Resultados: predominaram o atendimento de ocorrências clínicas, o sexo masculino e a média de idade de 55,5 anos. Após o atendimento pré-hospitalar, 50,2% dos pacientes permaneceram em observação e 34,8% precisaram de internação hospitalar. O tempo resposta pré-hospitalar apresentou correlação positiva com o tempo de internação. O principal desfecho foi a alta médica, sendo que 41% deles foram encaminhados para continuidade do tratamento. Conclusão: a análise mostrou um perfil de atendimentos a pacientes com idade média de 55 anos e acometidos por doenças de natureza clínica que necessitaram de encaminhamento a uma unidade de saúde, permanecendo em observação hospitalar por até 12h. Esses resultados são importantes para o embasamento de protocolos de fluxo assistencial na RUE, a fim de diminuir a sobrecarga dos serviços terciários.info:eu-repo/semantics/publishedVersio

Immobilization of trypsin in lignocellulosic waste material to produce peptides with bioactive potential from whey protein

ABSTRACT: In this study, trypsin (Enzyme Comission 3.4.21.4) was immobilized in a low cost, lignocellulosic support (corn cob powder—CCP) with the goal of obtaining peptides with bioactive potential from cheese whey. The pretreated support was activated with glyoxyl groups, glutaraldehyde and IDA-glyoxyl. The immobilization yields of the derivatives were higher than 83%, and the retention of catalytic activity was higher than 74%. The trypsin-glyoxyl-CCP derivative was thermally stable at 65 ̋C, a value that was 1090-fold higher than that obtained with the free enzyme. The trypsin-IDA-glyoxyl-CCP and trypsin-glutaraldehyde-CCP derivatives had thermal stabilities that were 883- and five-fold higher, respectively, then those obtained with the free enzyme. In the batch experiments, trypsin-IDA-glyoxyl-CCP retained 91% of its activity and had a degree of hydrolysis of 12.49%, while the values for trypsin-glyoxyl-CCP were 87% and 15.46%, respectively. The stabilized derivative trypsin-glyoxyl-CCP was also tested in an upflow packed-bed reactor. The hydrodynamic characterization of this reactor was a plug flow pattern, and the kinetics of this system provided a relative activity of 3.04 ̆ 0.01 U ̈ g ́1 and an average degree of hydrolysis of 23%, which were suitable for the production of potentially bioactive peptides

Hidrólise das proteínas do soro do queijo utilizando a alcalase imobilizada em pó de sabugo de milho / Hydrolysis of cheese whey proteins using immobilized alkali powder from corn cob

A hidrólise das proteínas do soro de queijo foi feita a 50º C, pH=9, 100 rpm por 24 h, utilizando o derivado alcalase glioxil pó de sabugo de milho (AGSM) e os resultados comparados com o derivado alcalase glioxil agarose (AGA) e enzima livre (AL). AL apresentou menor KM = 0,243mMe maior grau de hidrólise (DH)=59,63% com relação aos derivados, provavelmente devido aos efeitos difusionais da imobilização, dificultando o acesso do substrato a enzima. AGSM apresentou melhor estabilidade térmica (62 e 15,5 vezes maior do que a AL e AGA respectivamente). A SDS-PAGE demonstrou a ocorrência de hidrólise utilizando a AGSM, além do DH=26,59% e 3,29U/mg. Confirmando esta hidrólise, o perfil cromatográfico mostrou um aumento no número de picos em diferentes tempos de retenção. Resultados semelhantes foram encontrados para AGA, portanto, o SM representa uma fonte inovativa, de baixo custo, para imobilização da alcalase e obtenção de hidrolisados proteicos. 

Fatores relacionados às readmissões ao Serviço de Atendimento Móvel de Urgência

Resumo Objetivo analisar os fatores associados à readmissão de atendimento ao Serviço de Atendimento Móvel de Urgência (SAMU). Método estudo epidemiológico, do tipo seccional. Analisaram-se dados de 600 pacientes adultos atendidos pelo serviço de um município do interior de São Paulo, Brasil, no ano de 2015. Uma regressão logística múltipla identificou os fatores associados à readmissão. Resultados predominaram o atendimento de ocorrências clínicas, pacientes do sexo masculino e média de idade de 55,5 anos. Identificou-se um retorno de 26,7% nos seis meses seguintes ao atendimento no serviço pré-hospitalar. As readmissões se associaram aos fatores clínicos dos pacientes, aos procedimentos realizados no ambiente pré-hospitalar móvel e ao fluxo intra-hospitalar. Ademais, foi possível verificar relação com a região da cidade na qual o estudo foi realizado. Conclusão e implicações para a prática a análise mostrou um perfil de atendimentos a pacientes com idade média de 55 anos e acometidos por doenças crônicas não transmissíveis. A chance de retorno se associou à natureza clínica da doença, aos fluxos assistenciais e à região do atendimento. Estudos como este auxiliam no planejamento e na elaboração de políticas públicas e ações em saúde condizentes com as necessidades identificadas, com potencial de auxiliar na diminuição da sobrecarga dos serviços de urgência