{"id":4557,"date":"2021-04-29T10:10:14","date_gmt":"2021-04-29T10:10:14","guid":{"rendered":"https:\/\/longchangchemical.com\/?p=4557"},"modified":"2024-09-28T11:56:32","modified_gmt":"2024-09-28T11:56:32","slug":"immobilized-on-magnetic-silicon-based-materials","status":"publish","type":"post","link":"https:\/\/longchangchemical.com\/pt\/immobilized-on-magnetic-silicon-based-materials\/","title":{"rendered":"Par\u00e2metros experimentais da naringinase imobilizada em materiais magn\u00e9ticos \u00e0 base de sil\u00edcio"},"content":{"rendered":"<h1 class=\"wp-block-heading\"><strong>1. M\u00e9todo de detec\u00e7\u00e3o da atividade da enzima naringinase<\/strong><\/h1>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4583\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-45.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h3 class=\"wp-block-heading\">1-1 M\u00e9todo do espectrofot\u00f4metro<\/h3>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4582\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-44.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">O m\u00e9todo espectrofotom\u00e9trico do para-nitrofenol usa p-nitrofenol-\u03b1-ramnos\u00eddeo (pNP- \u03b1- Rha) e p-nitrofenol-\u03b2-D-glucopiranos\u00eddeo (pNP- \u03b2-Glu) como substrato, para detectar a atividade enzim\u00e1tica da \u03b1-ramnosidase e da \u03b2-glucosidase. Li Ping usou o m\u00e9todo do espectrofot\u00f4metro de p-nitrofenol, no experimento de adi\u00e7\u00e3o de \u03b2-glicosidase de Aspergillus niger na solu\u00e7\u00e3o de p-nitrofenol-\u03b2-glicos\u00eddeo (pNPG) para hidr\u00f3lise enzim\u00e1tica, e o valor da absorb\u00e2ncia foi medido no comprimento de onda ultravioleta de 400 nm. Com base no valor da absorb\u00e2ncia, calcula-se a atividade enzim\u00e1tica da \u03b2-glucosidase. Esse m\u00e9todo tem forte especificidade de substrato, mas o substrato padr\u00e3o \u00e9 caro, e a atividade enzim\u00e1tica da naringinase para hidrolisar glicos\u00eddeos em alimentos n\u00e3o pode ser determinada, portanto, esse m\u00e9todo n\u00e3o \u00e9 adequado para a detec\u00e7\u00e3o da atividade enzim\u00e1tica na prepara\u00e7\u00e3o industrial em larga escala da naringinase.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4581\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-43.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">O princ\u00edpio do m\u00e9todo do espectrofot\u00f4metro Davis \u00e9 que a naringina pode ocorrer em uma rea\u00e7\u00e3o cromog\u00eanica com dietilenoglicol 90% em condi\u00e7\u00f5es levemente alcalinas, e a produ\u00e7\u00e3o de chalcona amarela de naringina pode ser detectada sob luz ultravioleta com comprimento de onda de 420 nm. De acordo com o valor de absorb\u00e2ncia medido, a quantidade restante do substrato naringina pode ser calculada e, em seguida, a atividade da naringinase pode ser obtida por c\u00e1lculo. Wang Weina et al. usaram um m\u00e9todo Davis aprimorado, tomaram a naringina como substrato e adicionaram naringinase produzida por fermenta\u00e7\u00e3o l\u00edquida de Aspergillus niger FFCC 848 para hidr\u00f3lise enzim\u00e1tica. A absorb\u00e2ncia foi medida sob luz ultravioleta de 420 nm e, em seguida, calculada a atividade enzim\u00e1tica. O m\u00e9todo do espectrofot\u00f4metro Davis para medir a atividade da enzima \u00e9 simples de operar e tem uma velocidade r\u00e1pida, portanto, esse m\u00e9todo \u00e9 amplamente usado na detec\u00e7\u00e3o em tempo real da produ\u00e7\u00e3o de naringinase por fermenta\u00e7\u00e3o e da atividade da naringinase imobilizada.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4580\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-42.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h3 class=\"wp-block-heading\">1-2 Cromatografia l\u00edquida de alto desempenho<\/h3>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4579\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-41.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">O princ\u00edpio da cromatografia l\u00edquida de alto desempenho (HPLC) \u00e9 que a HPLC pode determinar com precis\u00e3o e analisar quantitativamente o substrato naringina, o produto intermedi\u00e1rio purunina e o produto final naringenina durante todo o processo de hidr\u00f3lise e, em seguida, calcular a atividade enzim\u00e1tica. Chen Yuelong e outros usaram a HPLC para determinar o tempo de reten\u00e7\u00e3o da naringina, arbutina, miricetina e seus produtos, e obtiveram uma separa\u00e7\u00e3o completa com uma resolu\u00e7\u00e3o superior a 1,5, monitorando com efic\u00e1cia o efeito da naringinase em v\u00e1rios glicos\u00eddeos no processo de hidr\u00f3lise e, ao mesmo tempo, a atividade da enzima foi determinada com precis\u00e3o. O m\u00e9todo HPLC tem as vantagens da alta precis\u00e3o e pode evitar a interfer\u00eancia de impurezas, mas, em compara\u00e7\u00e3o com o m\u00e9todo Davis, esse m\u00e9todo tem um tempo de detec\u00e7\u00e3o mais longo e uma opera\u00e7\u00e3o mais complicada.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4578\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-40.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h3 class=\"wp-block-heading\">1-3 M\u00e9todo aprimorado de determina\u00e7\u00e3o da atividade da enzima naringinase<\/h3>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4577\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-39.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">Usando o m\u00e9todo Davis aprimorado: 0,8 mL de 0,8 g-L<sup>-1<\/sup>\u00a0Uma solu\u00e7\u00e3o de naringina preparada com tamp\u00e3o de pH 4,5 foi adicionada ao sistema de rea\u00e7\u00e3o contendo 40 mg de naringinase imobilizada (ou 0,2 mL de naringinase livre). Coloque-o em um banho-maria com temperatura constante de 50 \u2103 por 30 minutos. Pegue 0,1 mL da solu\u00e7\u00e3o de hidr\u00f3lise enzim\u00e1tica da rea\u00e7\u00e3o e adicione 5 mL de dietilenoglicol (fra\u00e7\u00e3o volum\u00e9trica de 90%) e 0,1 mL de solu\u00e7\u00e3o de NaOH (4 mol-L<sup>-1<\/sup>) em tubos e deixe em repouso por 10 minutos ap\u00f3s mistur\u00e1-los uniformemente. Me\u00e7a a absorb\u00e2ncia da solu\u00e7\u00e3o misturada a 420 nm.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4576\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-38.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">A defini\u00e7\u00e3o da atividade da enzima naringinase (U): em uma condi\u00e7\u00e3o de pH 4,5, 50 \u2103, 1 unidade de atividade enzim\u00e1tica \u00e9 a quantidade de enzima necess\u00e1ria para degradar 1 \u03bcg de naringina por minuto. Taxa de atividade enzim\u00e1tica ( U-g<sup>-1<\/sup>\u00a0ou U-mL<sup>-1<\/sup>) \u00e9 definida como: na condi\u00e7\u00e3o de pH 4,5, 50 \u2103, a atividade enzim\u00e1tica exibida por 1 g de naringinase imobilizada (ou 1 mL de naringinase livre). A atividade enzim\u00e1tica relativa \u00e9 calculada de acordo com a f\u00f3rmula a seguir:<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4575\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-37.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">Atividade enzim\u00e1tica relativa \/% = atividade enzim\u00e1tica\/atividade enzim\u00e1tica m\u00e1xima\u00d7100<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4574\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-36.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">Determina\u00e7\u00e3o da curva padr\u00e3o: a concentra\u00e7\u00e3o de massa da solu\u00e7\u00e3o padr\u00e3o de glicos\u00eddeo naringina est\u00e1 disposta em 0, 0,4, 0,8, 1,2, 1,6 g - L<sup>-1<\/sup>e, em seguida, 0,1 mL da solu\u00e7\u00e3o foi levado para rea\u00e7\u00e3o com 5 mL de dietilenoglicol (90% , v \/ v) e 0,1 mL de solu\u00e7\u00e3o de hidr\u00f3xido de s\u00f3dio ( 4 mol-L<sup>-1<\/sup>). Depois que a mistura foi homog\u00eanea e permaneceu em repouso por 10 minutos, o valor de absor\u00e7\u00e3o de luz da solu\u00e7\u00e3o mista foi medido a 420 nm, e a curva padr\u00e3o foi desenhada de acordo.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4573\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-35.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h2 class=\"wp-block-heading\"><strong>2. Prepara\u00e7\u00e3o da solu\u00e7\u00e3o de enzima naringinase<\/strong><\/h2>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4572\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-34.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">O Aspergillus niger FFCC uv-11 armazenado a 4 \u2103 foi transferido para o meio slant, e os esporos maduros foram obtidos por meio de cultura em temperatura constante a 30 \u2103 por 96 h. Em seguida, os esporos foram lavados com solu\u00e7\u00e3o salina normal est\u00e9ril com uma fra\u00e7\u00e3o de massa de 0,9%. O valor da absorb\u00e2ncia da suspens\u00e3o de esporos \u00e9 medido a 600 nm (valor OD600), com solu\u00e7\u00e3o salina est\u00e9ril seu valor OD600 foi ajustado para 0,200. Em seguida, a suspens\u00e3o de esporos com a fra\u00e7\u00e3o de volume de 10% foi inoculada em 100 mL de meio de fermenta\u00e7\u00e3o (frasco c\u00f4nico de 250 mL). O caldo de fermenta\u00e7\u00e3o obtido foi centrifugado a 4 \u2103 e 6000 r-min<sup>-1<\/sup>\u00a0por 10 minutos em uma centr\u00edfuga refrigerada. Em seguida, passe pelo filtro de suc\u00e7\u00e3o para obter o fluido sobrenadante necess\u00e1rio para a enzima naringina. Refrigere-o a 4 \u2103 para uso posterior.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4571\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-33.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h2 class=\"wp-block-heading\"><strong>3. Prepara\u00e7\u00e3o da enzima naringinase imobilizada em quitosana<\/strong><\/h2>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4570\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-32.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">Tomamos 3,0 g de quitosana e dissolvemos 100 mL de fra\u00e7\u00e3o volum\u00e9trica da solu\u00e7\u00e3o de \u00e1cido ac\u00e9tico 2% para preparar 0,3 g - L<sup>-1<\/sup>\u00a0A solu\u00e7\u00e3o coloidal de quitosana foi submetida a ultrassom por 20 minutos para remover as bolhas de ar e permitir que ela se dissolvesse completamente. A solu\u00e7\u00e3o coloidal de quitosana foi injetada gota a gota com uma agulha em 0,5 g - L<sup>-1<\/sup>\u00a0NaOH condensado (1000 mL) para formar uma pelota lisa. Deixe repousar em temperatura ambiente por 3 horas, e os gr\u00e2nulos foram lavados repetidamente com \u00e1gua destilada at\u00e9 ficarem neutros, e a \u00e1gua foi absorvida por um papel de filtro para obter as microesferas de quitosana. Foram pesadas 0,4 g de microesferas de quitosana, foram adicionados 0,4 ml de glutaralde\u00eddo com fra\u00e7\u00e3o de volume 6% e as microesferas foram lavadas repetidamente com \u00e1gua destilada at\u00e9 que n\u00e3o houvesse glutaralde\u00eddo na superf\u00edcie das microesferas ap\u00f3s temperatura constante e vibra\u00e7\u00e3o a 25 \u2103 150 r-min.<sup>-1<\/sup>\u00a0As microesferas de quitosana com liga\u00e7\u00f5es cruzadas foram obtidas por secagem com papel de filtro.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4569\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-31.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">\u00a0Ele disse que 0,4 g de microesferas de quitosana, adicionar 0,4 mL de n\u00famero de membro de cr\u00e9ditos 6% glutaralde\u00eddo, a 150 r - min<sup>-1<\/sup>\u00a0Depois de 2 horas, lavadas repetidamente com \u00e1gua destilada, as microesferas n\u00e3o s\u00e3o pentil dialde\u00eddo e o papel de filtro \u00e9 seco para obter microesferas de quitosana reticuladas. Pegue 4 mL de solu\u00e7\u00e3o de enzima naringinase, adicione-a a 0,4 g de microesferas de quitosana reticuladas, agite a uma temperatura constante de 25 \u2103, 150 r-min<sup>-1<\/sup>\u00a0por 2 h, e coloque-o em um refrigerador a 4 \u2103 para acoplamento de adsor\u00e7\u00e3o por 4 h. Enx\u00e1gue a solu\u00e7\u00e3o enzim\u00e1tica residual na superf\u00edcie do transportador com tamp\u00e3o de pH 4,5, absorva a \u00e1gua no papel de filtro para obter a naringinase imobilizada e armazene-a em um refrigerador a 4 \u2103 para uso posterior.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4568\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-30.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h2 class=\"wp-block-heading\"><strong>4. Prepara\u00e7\u00e3o de microesferas magn\u00e9ticas de quitosana \u00e0 base de sil\u00edcio (MSC)<\/strong><\/h2>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4567\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-29.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">O processo de prepara\u00e7\u00e3o de nanopart\u00edculas magn\u00e9ticas \u00e0 base de sil\u00edcio (MS) \u00e9 mostrado na Figura 1.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" width=\"554\" height=\"175\" class=\"wp-image-4559\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-21.png\" alt=\"\" srcset=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-21.png 554w, https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-21-300x95.png 300w\" sizes=\"(max-width: 554px) 100vw, 554px\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">Figura 1 Diagrama esquem\u00e1tico da prepara\u00e7\u00e3o de nanopart\u00edculas magn\u00e9ticas \u00e0 base de sil\u00edcio<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4566\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-28.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">( 1) Primeiro, o Fe<sub>3<\/sub>O<sub>4<\/sub>\u00a0modificadas com \u00e1cido c\u00edtrico foram preparadas pelo m\u00e9todo de co-precipita\u00e7\u00e3o qu\u00edmica, e o procedimento experimental que compreende: 2,25 g de FeCl<sub>3<\/sub>-6H<sub>2<\/sub>O e 1,61 g de FeSO<sub>4<\/sub>-7H<sub>2<\/sub>O s\u00e3o adicionados a um bal\u00e3o de tr\u00eas gargalos preenchido com 100 mL de \u00e1gua deionizada, a temperatura do sistema foi aquecida a 85 \u2103 sob a condi\u00e7\u00e3o de g\u00e1s nitrog\u00eanio. Agite sob 1000 r-min<sup>-1<\/sup>\u00a0e despeje rapidamente 10 mL de \u00e1gua de am\u00f4nia concentrada. Depois que a solu\u00e7\u00e3o ficar preta, adicione 153 mg de citrato de s\u00f3dio. Ap\u00f3s 30 minutos de rea\u00e7\u00e3o, separe a produ\u00e7\u00e3o com um \u00edm\u00e3 e lave a fase s\u00f3lida com \u00e1gua deionizada e etanol v\u00e1rias vezes. O Fe<sub>3<\/sub>O<sub>4<\/sub>\u00a0As nanopart\u00edculas obtidas s\u00e3o secas para uso posterior.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4565\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-27.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">( 2) Em seguida, nanopart\u00edculas magn\u00e9ticas \u00e0 base de sil\u00edcio (MS) foram preparadas pelo m\u00e9todo sol-gel, mostrado na Figura 1. As etapas experimentais incluem: 0,1 g de Fe<sub>3<\/sub>O<sub>4<\/sub>\u00a0foram uniformemente dispersas em uma solu\u00e7\u00e3o mista contendo 40 mL de etanol anidro, 10 mL de \u00e1gua destilada e 1,2 mL de \u00e1gua de am\u00f4nia concentrada. Sob a condi\u00e7\u00e3o de agita\u00e7\u00e3o de 300 r-min<sup>-1<\/sup>, got\u00edculas de solu\u00e7\u00e3o contendo Fe<sub>3<\/sub>O<sub>4<\/sub>\u00a0foram adicionadas ao TEOS (0,4 mL) disperso em 30 mL de etanol anidro, e a rea\u00e7\u00e3o das nanopart\u00edculas \u00e0 base de sil\u00edcio foi realizada durante 12 horas em temperatura ambiente. Os produtos foram lavados com etanol anidro e \u00e1gua destilada por v\u00e1rias vezes para obter nanopart\u00edculas magn\u00e9ticas \u00e0 base de s\u00edlica (MS), que foram secas para uso posterior.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4564\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-26.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">As microesferas de MSC foram preparadas pelo m\u00e9todo de reticula\u00e7\u00e3o de emuls\u00e3o, conforme mostrado na Figura 2. As etapas experimentais inclu\u00edram: 0,125 g de nanopart\u00edculas de s\u00edlica magn\u00e9tica (MS) uniformemente dispersas em 10 mL de solu\u00e7\u00e3o de quitosana com fra\u00e7\u00e3o de massa de 2,5% preparada por quitosana dissolvida em solu\u00e7\u00e3o de \u00e1cido ac\u00e9tico com fra\u00e7\u00e3o de volume de 5,0% e, em seguida, adicione a solu\u00e7\u00e3o de quitosana em uma emuls\u00e3o contendo 60 mL de cera de parafina l\u00edquida e 1,8 mL de twain 80, gota a gota. Depois de agitar por 30 minutos em uma condi\u00e7\u00e3o de banho-maria de 40 \u2103 e 800 r-min<sup>-1<\/sup>\uff0cadicione 2,0 mL de glutaralde\u00eddo e reaja por 1 h. Em seguida, o pH do sistema foi ajustado para 9,0 com 1 mol-L<sup>-1<\/sup>\u00a0NaOH, e a rea\u00e7\u00e3o foi continuada por 1 h. Os produtos foram lavados com \u00e9ter de petr\u00f3leo, etanol e \u00e1gua destilada v\u00e1rias vezes para obter microesferas de MSC, que foram secas a v\u00e1cuo para uso posterior.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" width=\"554\" height=\"246\" class=\"wp-image-4560\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-22.png\" alt=\"\" srcset=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-22.png 554w, https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-22-300x133.png 300w\" sizes=\"(max-width: 554px) 100vw, 554px\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">Figura 2 Diagrama esquem\u00e1tico da prepara\u00e7\u00e3o de microesferas magn\u00e9ticas de quitosana \u00e0 base de sil\u00edcio<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4584\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-46.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h2 class=\"wp-block-heading\"><strong>5. Prepara\u00e7\u00e3o de microesferas magn\u00e9ticas de quitosana \u00e0 base de sil\u00edcio epox\u00eddico (MSCE)<\/strong><\/h2>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4585\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-47.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">0,5 g de microesferas secas de MSC preparadas pelo m\u00e9todo 4 foram pesadas e adicionadas a um frasco c\u00f4nico de 50 mL. Em seguida, 2,0 mL de solu\u00e7\u00e3o de NaOH, solu\u00e7\u00e3o de epicrolohidrina e NaBH<sub>4<\/sub>\u00a0foram adicionados ao frasco com uma determinada concentra\u00e7\u00e3o, respectivamente. E a rea\u00e7\u00e3o foi realizada por oscila\u00e7\u00e3o de temperatura constante por um determinado tempo. O mecanismo de rea\u00e7\u00e3o da prepara\u00e7\u00e3o do MSCE \u00e9 mostrado na Figura 3.<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" width=\"554\" height=\"179\" class=\"wp-image-4561\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-23.png\" alt=\"\" srcset=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-23.png 554w, https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-23-300x97.png 300w\" sizes=\"(max-width: 554px) 100vw, 554px\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">Figura 3 Princ\u00edpio de prepara\u00e7\u00e3o de microesferas magn\u00e9ticas de quitosana \u00e0 base de silicone com base em ep\u00f3xi<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4586\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-48.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">Ap\u00f3s a ativa\u00e7\u00e3o, as microesferas ativadas s\u00e3o lavadas repetidamente com \u00e1gua destilada at\u00e9 que n\u00e3o haja grupos OH- e ep\u00f3xi livres na superf\u00edcie para obter microesferas magn\u00e9ticas de quitosana \u00e0 base de silicone \u00e0 base de ep\u00f3xi (MSCE). O m\u00e9todo de detec\u00e7\u00e3o \u00e9 o seguinte: pegue 3,0 mL do sobrenadante da lavagem e adicione de 1 a 2 gotas de fenolftale\u00edna. Depois de agitar, se n\u00e3o ficar vermelho, significa que n\u00e3o h\u00e1 OH- livre com MSCE; pegue 3,0 mL do sobrenadante da lavagem e adicione 3,0 mL de 1,3 mol-L<sup>-1<\/sup>\u00a0tiossulfato de s\u00f3dio e 1 a 2 gotas de fenolftale\u00edna. Se a solu\u00e7\u00e3o n\u00e3o ficar vermelha ap\u00f3s a agita\u00e7\u00e3o, significa que n\u00e3o h\u00e1 grupos ep\u00f3xi livres no MSCE. A equa\u00e7\u00e3o da rea\u00e7\u00e3o de detec\u00e7\u00e3o do grupo ep\u00f3xi \u00e9 a seguinte<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" width=\"554\" height=\"83\" class=\"wp-image-4562\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-24.png\" alt=\"\" srcset=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-24.png 554w, https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-24-300x45.png 300w\" sizes=\"(max-width: 554px) 100vw, 554px\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\"><strong>6 Prepara\u00e7\u00e3o da enzima naringinase imobilizada em MSCE<\/strong><\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" class=\"wp-image-4587\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-49.png\" alt=\"\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">5,0 g de MSCE preparado pelo m\u00e9todo 5 foram adicionados a 50 mL de solu\u00e7\u00e3o de enzima naringinase com um determinado pH, e a rea\u00e7\u00e3o foi oscilada em um banho-maria de temperatura constante, conforme mostrado na Figura 4. Ap\u00f3s a rea\u00e7\u00e3o, as microesferas foram lavadas com solu\u00e7\u00e3o tamp\u00e3o v\u00e1rias vezes at\u00e9 que nenhuma naringinase livre fosse encontrada na superf\u00edcie das microesferas. As microesferas foram drenadas por um funil de Brinella para obter a enzima naringinase imobilizada MSCE, que foi refrigerada a 4 \u2103 para uso posterior.\u00a0<\/h4>\r\n\r\n\r\n\r\n<figure class=\"wp-block-image size-large\"><img decoding=\"async\" width=\"554\" height=\"183\" class=\"wp-image-4563\" src=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-25.png\" alt=\"\" srcset=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-25.png 554w, https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-25-300x99.png 300w\" sizes=\"(max-width: 554px) 100vw, 554px\" \/><\/figure>\r\n\r\n\r\n\r\n<h4 class=\"wp-block-heading\">Figura 4 Princ\u00edpio de prepara\u00e7\u00e3o da enzima naringinase imobilizada em MSCE<\/h4>\r\n\r\n\r\n\r\n<h2 class=\"wp-block-heading\"><strong><b>Entre em contato conosco agora!<\/b><\/strong><\/h2>\r\n<h4><strong><b>Se precisar do Price, preencha suas informa\u00e7\u00f5es de contato no formul\u00e1rio abaixo. Normalmente, entraremos em contato dentro de 24 horas. Voc\u00ea tamb\u00e9m pode me enviar um e-mail\u00a0<span style=\"color: #00ccff;\"><a style=\"color: #00ccff;\" href=\"mailto:info@longchangchemical.com\">info@longchangchemical.com<\/a><\/span>\u00a0durante o hor\u00e1rio comercial (das 8h30 \u00e0s 18h UTC+8 de segunda a s\u00e1bado) ou use o bate-papo ao vivo do site para obter uma resposta imediata.<\/b><\/strong><\/h4>\r\n<table style=\"border-collapse: collapse; width: 326.27pt;\" border=\"0\" width=\"435\" cellspacing=\"0\" cellpadding=\"0\">\r\n<tbody>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt; width: 164.25pt;\" width=\"219\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/compound-glucoamylase-cas-9032-08-0\/\"><span style=\"color: #00ccff;\">Composto Glucoamilase<\/span><\/a><\/td>\r\n<td class=\"et2\" style=\"width: 162.00pt;\" width=\"216\">9032-08-0<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/pullulanase-cas-9075-68-7\/\"><span style=\"color: #00ccff;\">Pullulanase<\/span><\/a><\/td>\r\n<td class=\"et2\">9075-68-7<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/xylanase-cas-37278-89-0\/\"><span style=\"color: #00ccff;\">Xilanase<\/span><\/a><\/td>\r\n<td class=\"et2\">37278-89-0<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/cellulase-cas-9012-54-8\/\"><span style=\"color: #00ccff;\">Celulase<\/span><\/a><\/td>\r\n<td class=\"et2\">9012-54-8<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/naringinase-cas-9068-31-9\/\"><span style=\"color: #00ccff;\">Naringinase<\/span><\/a><\/td>\r\n<td class=\"et2\">9068-31-9<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/beta-amylase-cas-9000-91-3\/\"><span style=\"color: #00ccff;\">\u03b2-Amilase<\/span><\/a><\/td>\r\n<td class=\"et2\">9000-91-3<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/glucose-oxidase-cas-9001-37-0\/\"><span style=\"color: #00ccff;\">Glucose oxidase<\/span><\/a><\/td>\r\n<td class=\"et2\">9001-37-0<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\">alfa-Amilase<\/td>\r\n<td class=\"et2\">9000-90-2<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/longzyme-acid-pectinase-cas-9032-75-1\/\"><span style=\"color: #00ccff;\">Pectinase<\/span><\/a><\/td>\r\n<td class=\"et2\">9032-75-1<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\">Peroxidase<\/td>\r\n<td class=\"et2\">9003-99-0<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/lipase-cas-9001-62-1\/\"><span style=\"color: #00ccff;\">Lipase<\/span><\/a><\/td>\r\n<td class=\"et2\">9001-62-1<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/catalase-cas-9001-05-2\/\"><span style=\"color: #00ccff;\">Catalase<\/span><\/a><\/td>\r\n<td class=\"et4\">9001-05-2<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/tannase-cas-9025-71-2\/\"><span style=\"color: #00ccff;\">TANNASE<\/span><\/a><\/td>\r\n<td class=\"et2\">9025-71-2<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/elastase-cas-39445-21-1\/\"><span style=\"color: #00ccff;\">Elastase<\/span><\/a><\/td>\r\n<td class=\"et2\">39445-21-1<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/urease-cas-9002-13-5\/\"><span style=\"color: #00ccff;\">Urease<\/span><\/a><\/td>\r\n<td class=\"et2\">9002-13-5<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/dextranase-cas-9025-70-1\/\"><span style=\"color: #00ccff;\">DEXTRANASE<\/span><\/a><\/td>\r\n<td class=\"et2\">9025-70-1<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.5pt; text-align: left;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/l-lactic-dehydrogenase-cas-9001-60-9\/\"><span style=\"color: #00ccff;\">L-L\u00e1ctico desidrogenase<\/span><\/a><\/td>\r\n<td class=\"et2\">9001-60-9<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/dehydrogenase-malate-cas-9001-64-3\/\"><span style=\"color: #00ccff;\">Malato desidrogenase<\/span><\/a><\/td>\r\n<td class=\"et2\">9001-64-3<\/td>\r\n<\/tr>\r\n<tr style=\"height: 13.50pt;\">\r\n<td class=\"et2\" style=\"height: 13.50pt;\" height=\"18\"><a href=\"https:\/\/longchangchemical.com\/pt\/product\/cholesterol-oxidase-cas-9028-76-6\/\"><span style=\"color: #00ccff;\">Colesterol oxidase<\/span><\/a><\/td>\r\n<td class=\"et2\">9028-76-6<\/td>\r\n<\/tr>\r\n<\/tbody>\r\n<\/table>","protected":false},"excerpt":{"rendered":"<p>1. M\u00e9todo de detec\u00e7\u00e3o da atividade enzim\u00e1tica da naringinase 1-1 M\u00e9todo do espectrofot\u00f4metro O m\u00e9todo espectrofotom\u00e9trico do para-nitrofenol usa o p-nitrofenol-\u03b1-ramnos\u00eddeo (pNP-\u03b1-Rha) e o p-nitrofenol-\u03b2-D-glucopiranos\u00eddeo (pNP-\u03b2-Glu) como substrato para detectar a atividade enzim\u00e1tica da \u03b1-ramnosidase e da \u03b2-glucosidase. Li Ping usou o m\u00e9todo do espectrofot\u00f4metro de p-nitrofenol, no experimento de adi\u00e7\u00e3o de \u03b2-glicosidase de Aspergillus niger na solu\u00e7\u00e3o de p-nitrofenol -\u03b2-glicos\u00eddeo ( pNPG ) para hidr\u00f3lise enzim\u00e1tica, e o valor da absorb\u00e2ncia foi medido em [...]<\/p>","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[108],"tags":[],"class_list":["post-4557","post","type-post","status-publish","format-standard","hentry","category-enzyme-news"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v25.3.1 - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>Experimental parameters of\u00a0naringinase immobilized on magnetic silicon based materials - Longchang Chemical<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/longchangchemical.com\/pt\/immobilized-on-magnetic-silicon-based-materials\/\" \/>\n<meta property=\"og:locale\" content=\"pt_BR\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Experimental parameters of\u00a0naringinase immobilized on magnetic silicon based materials - Longchang Chemical\" \/>\n<meta property=\"og:description\" content=\"1.\u00a0Detection method of naringinase enzyme activity 1-1\u00a0Spectrophotometer method The para nitrophenol spectrophotometric method\u00a0uses p-nitrophenol-\u00a0\u03b1-rhamnoside (\u00a0pNP-\u00a0\u03b1-\u00a0Rha\u00a0) and p-nitrophenol-\u03b2-D-glucopyranoside (\u00a0pNP-\u00a0\u03b2-Glu) as substrate, for detecting the enzyme activity of\u00a0the \u03b1-rhamnosidase\u00a0and \u03b2-glucosidase.\u00a0Li Ping\u00a0used the p-nitrophenol spectrophotometer method, in the experiment of\u00a0adding\u00a0Aspergillus niger \u03b2-glucosidase into the solution of\u00a0p-nitrophenol\u00a0-\u03b2-glucoside (\u00a0pNPG\u00a0) for enzymatic hydrolysis, and the value of absorbance was measured at [&hellip;]\" \/>\n<meta property=\"og:url\" content=\"https:\/\/longchangchemical.com\/pt\/immobilized-on-magnetic-silicon-based-materials\/\" \/>\n<meta property=\"og:site_name\" content=\"Longchang Chemical\" \/>\n<meta property=\"article:published_time\" content=\"2021-04-29T10:10:14+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2024-09-28T11:56:32+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-45.png\" \/>\n<meta name=\"author\" content=\"Longchang Chemical\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\/\/longchangchemical.com\/immobilized-on-magnetic-silicon-based-materials\/#article\",\"isPartOf\":{\"@id\":\"https:\/\/longchangchemical.com\/immobilized-on-magnetic-silicon-based-materials\/\"},\"author\":{\"name\":\"Longchang Chemical\",\"@id\":\"https:\/\/longchangchemical.com\/#\/schema\/person\/ecb1ad846136c6f1ca534b62978c6a8e\"},\"headline\":\"Experimental parameters of\u00a0naringinase immobilized on magnetic silicon based materials\",\"datePublished\":\"2021-04-29T10:10:14+00:00\",\"dateModified\":\"2024-09-28T11:56:32+00:00\",\"mainEntityOfPage\":{\"@id\":\"https:\/\/longchangchemical.com\/immobilized-on-magnetic-silicon-based-materials\/\"},\"wordCount\":1840,\"commentCount\":0,\"publisher\":{\"@id\":\"https:\/\/longchangchemical.com\/#organization\"},\"image\":{\"@id\":\"https:\/\/longchangchemical.com\/immobilized-on-magnetic-silicon-based-materials\/#primaryimage\"},\"thumbnailUrl\":\"https:\/\/longchangchemical.com\/wp-content\/uploads\/2021\/04\/image-45.png\",\"articleSection\":[\"Enzyme News\"],\"inLanguage\":\"pt-BR\",\"potentialAction\":[{\"@type\":\"CommentAction\",\"name\":\"Comment\",\"target\":[\"https:\/\/longchangchemical.com\/immobilized-on-magnetic-silicon-based-materials\/#respond\"]}]},{\"@type\":\"WebPage\",\"@id\":\"https:\/\/longchangchemical.com\/immobilized-on-magnetic-silicon-based-materials\/\",\"url\":\"https:\/\/longchangchemical.com\/immobilized-on-magnetic-silicon-based-materials\/\",\"name\":\"Experimental parameters of\u00a0naringinase immobilized on magnetic silicon based materials - 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