{"id":7270,"date":"2024-08-27T14:10:59","date_gmt":"2024-08-27T14:10:59","guid":{"rendered":"https:\/\/longchangchemical.com\/?p=7270"},"modified":"2024-09-28T11:52:28","modified_gmt":"2024-09-28T11:52:28","slug":"modification-of-protease-producing-strains","status":"publish","type":"post","link":"https:\/\/longchangchemical.com\/pt\/modification-of-protease-producing-strains\/","title":{"rendered":"Modifica\u00e7\u00e3o de cepas produtoras de protease"},"content":{"rendered":"

Modifica\u00e7\u00e3o de cepas produtoras de protease<\/strong><\/h1>\n

A protease \u00e9 uma classe de enzima que catalisa a hidr\u00f3lise de liga\u00e7\u00f5es pept\u00eddicas em prote\u00ednas, amplamente encontrada em animais, plantas e microrganismos, e \u00e9 uma das prepara\u00e7\u00f5es enzim\u00e1ticas mais usadas em esp\u00e9cies de enzimas industriais, representando cerca de 60% da quantidade total de enzimas. As fontes microbianas de proteases s\u00e3o abundantes, diversificadas e f\u00e1ceis de produzir, tornando-se a principal fonte de proteases no mercado. A produ\u00e7\u00e3o comercial de protease pode ser rastreada at\u00e9 o in\u00edcio do s\u00e9culo passado. Em 1908, os alem\u00e3es come\u00e7aram a usar a enzima pancre\u00e1tica para curtimento de couro; em 1911, a empresa Wallerstein dos Estados Unidos produziu papa\u00edna como agente clarificador para cerveja; no in\u00edcio dos anos 60, a Holanda produziu detergentes com a adi\u00e7\u00e3o de protease. Al\u00e9m disso, tamb\u00e9m combinada com a pr\u00e1tica para realizar a sele\u00e7\u00e3o e a cria\u00e7\u00e3o de bact\u00e9rias resistentes ao calor, aos \u00e1cidos e aos sais para a produ\u00e7\u00e3o de protease, bem como o petr\u00f3leo como mat\u00e9ria-prima para a produ\u00e7\u00e3o de fermenta\u00e7\u00e3o da pesquisa de protease alcalina.<\/p>\n

 <\/p>\n

H\u00e1 muitos tipos de proteases, e seu peso molecular, tamanho, estrutura espacial e fun\u00e7\u00e3o s\u00e3o muito diferentes, mas cada fam\u00edlia de proteases ainda mant\u00e9m um dom\u00ednio estrutural altamente conservado. Atualmente, mais de 100 tipos de proteases foram cristalizadas ou altamente purificadas, e muitas delas foram elucidadas em termos de suas estruturas prim\u00e1rias e est\u00e9reo (estrutura terci\u00e1ria).<\/p>\n

 <\/p>\n

 <\/p>\n

2 Modifica\u00e7\u00e3o de cepas de protease<\/p>\n

A constru\u00e7\u00e3o de bact\u00e9rias de engenharia para a express\u00e3o eficiente de protease e a otimiza\u00e7\u00e3o de suas propriedades enzim\u00e1ticas t\u00eam sido o ponto alto de acad\u00eamicos nacionais e estrangeiros, e os meios normalmente usados s\u00e3o: reprodu\u00e7\u00e3o por muta\u00e7\u00e3o, fus\u00e3o protoplasm\u00e1tica, constru\u00e7\u00e3o de bact\u00e9rias geneticamente modificadas e tecnologia de evolu\u00e7\u00e3o direcional in vitro.<\/p>\n

 <\/p>\n

2.1 Mutag\u00eanese, reprodu\u00e7\u00e3o e fus\u00e3o protoplasm\u00e1tica<\/p>\n

A reprodu\u00e7\u00e3o por mutag\u00eanese ocorre principalmente por meio da mutag\u00eanese por radia\u00e7\u00e3o ou de alguns reagentes qu\u00edmicos para induzir muta\u00e7\u00f5es no material gen\u00e9tico da bact\u00e9ria a fim de obter cepas mutantes com excelentes caracter\u00edsticas. Por exemplo, ap\u00f3s a mutag\u00eanese por UV de cepas produtoras de enzimas por Cai Wanling et al., sua atividade enzim\u00e1tica aumentou em 16% em compara\u00e7\u00e3o com a cepa original. Yanli Li et al. examinaram a atividade da enzima produtora de protease neutra do Aspergillus oryzae ZW-06 at\u00e9 15.000 U\/g de coalhada seca por mutag\u00eanese dirigida por Co60, que foi 74% maior do que antes da mutag\u00eanese. Huang Hongying et al. combinaram a tecnologia de implanta\u00e7\u00e3o de \u00edons N+ de baixa energia no Bacillus licheniformis do tipo selvagem por quatro vezes com diferentes meios f\u00edsico-qu\u00edmicos de mutag\u00eanese, para obter uma cepa de alto rendimento, a atividade enzim\u00e1tica bruta de 12425. 9 U\/mL, 17,1 vezes maior do que a cepa inicial.<\/p>\n

 <\/p>\n

 <\/p>\n

A tecnologia de fus\u00e3o protoplasm\u00e1tica \u00e9 um processo no qual as c\u00e9lulas de uma linhagem biparental s\u00e3o desparafinadas para fus\u00e3o protoplasm\u00e1tica, de modo a trocar e recombinar seus genomas para obter recombinantes est\u00e1veis. Por exemplo, Pan Yanyun et al. usaram o m\u00e9todo de fus\u00e3o protoplasm\u00e1tica para fundir Bacillus licheniformis 2709 com Bacillus subtilis BD105, que cont\u00e9m o vetor de clonagem do gene da protease alcalina pDW2, para obter uma cepa de alto rendimento A16, com produ\u00e7\u00e3o de enzima de fermenta\u00e7\u00e3o 50%-100% maior do que a de 2709 e produ\u00e7\u00e3o de enzima de at\u00e9 30.000 U\/mL no teste de frasco agitado.<\/p>\n

 <\/p>\n

2.2 Constru\u00e7\u00e3o de bact\u00e9rias de engenharia gen\u00e9tica<\/p>\n

A constru\u00e7\u00e3o de bact\u00e9rias de engenharia geralmente requer hospedeiros de express\u00e3o e vetores de express\u00e3o ou elementos de express\u00e3o adequados. Primeiramente, o gene-alvo \u00e9 clonado no vetor de express\u00e3o para obter o vetor recombinante, depois transformado no hospedeiro de express\u00e3o, passando pelo processo de triagem e, por fim, obtendo o processo da cepa de alto rendimento. Tamb\u00e9m \u00e9 poss\u00edvel integrar os fragmentos ex\u00f3genos do gene-alvo diretamente no genoma do hospedeiro para obter cepas de alto rendimento.<\/p>\n

 <\/p>\n

[18-20 de outubro] 2024 A 2\u00aa Confer\u00eancia sobre Engenharia Avan\u00e7ada de Enzimas e Aplica\u00e7\u00f5es de Tecnologia de Enzimas<\/p>\n

Min Zhang et al. ligaram a sequ\u00eancia do pept\u00eddeo sinalizador do promotor (sacR) do gene sacB com o gene da protease neutra do Bacillus subtilis e o clonaram no vetor pHP13, constru\u00edram o vetor de secre\u00e7\u00e3o de express\u00e3o induz\u00edvel pHP13SN contendo o gene da protease neutra e o transformaram no Bacillus subtilis DB104, que apresentou um aumento de 48% na viabilidade da enzima em compara\u00e7\u00e3o com a cepa de partida. wangHui et al. clonaram a Banpr, uma protease neutra do Bacillus amyloliquefaciens K11, e constru\u00edram um plasm\u00eddeo de express\u00e3o recombinante, pUB110-Banpr, com Bacillus amyloliquefaciens K11 como hospedeiro de express\u00e3o, e a atividade enzim\u00e1tica na fermenta\u00e7\u00e3o em frasco agitado atingiu 8995 \u00b1 250 U\/mL e 28084 \u00b1 1282 U\/mL em um sistema de fermenta\u00e7\u00e3o de 15 L, muito mais do que a da cepa industrial Bacillus subtilis AS.1398 (8000-10000 U\/mL).<\/p>\n

 <\/p>\n

2.3 Melhoria das propriedades da protease<\/p>\n

Com o desenvolvimento de t\u00e9cnicas de evolu\u00e7\u00e3o dirigida in vitro, \u00e9 poss\u00edvel obter prote\u00ednas com fun\u00e7\u00f5es aprimoradas ou novas por meio de muta\u00e7\u00e3o aleat\u00f3ria de genes codificadores, t\u00e9cnicas de DNAShuffling e triagem dirigida sem o conhecimento das informa\u00e7\u00f5es estruturais tridimensionais e do mecanismo de a\u00e7\u00e3o da prote\u00edna-alvo.<\/p>\n

 <\/p>\n

 <\/p>\n

A estabilidade da enzima \u00e9 um fator fundamental para o sucesso da comercializa\u00e7\u00e3o. A estabilidade da enzima pode ser melhorada com a introdu\u00e7\u00e3o de pontes de sal, liga\u00e7\u00f5es dissulfeto, intera\u00e7\u00f5es arom\u00e1ticas e aumento da afinidade da enzima por \u00edons de c\u00e1lcio, aumentando assim a rigidez da mol\u00e9cula por meio de mutag\u00eanese direcionada. A substitui\u00e7\u00e3o de Gly61 ou Ser98 da protease BPN' por Cys, e de ambos, resultou em um aumento na estabilidade t\u00e9rmica da enzima sem altera\u00e7\u00e3o na efici\u00eancia catal\u00edtica.<\/p>\n

 <\/p>\n

Ao substituir Val72 por Ile, Ala92 por Thr e Gly131 por Asp da protease de B. subtilis, a enzima mutante pode catalisar a atividade do substrato 2 vezes maior do que a da enzima selvagem a 10\u00b0C. Em 2005, a empresa dinamarquesa Novozymes introduziu uma protease de baixa temperatura, a \"Polarzyme\", que pode ser adicionada ao detergente. Ao adicion\u00e1-la ao detergente, a temperatura de lavagem diminuiu de 60\u00b0C e 40\u00b0C para 30\u00b0C e 20\u00b0C, economizando 60% de energia el\u00e9trica, e o volume de vendas do detergente \"carecoldwash\" com protease de baixa temperatura aumentou sete vezes.<\/p>\n

 <\/p>\n

3 Perspectivas<\/p>\n

As proteases t\u00eam sido usadas em diversos campos que est\u00e3o intimamente relacionados \u00e0 nossa vida cotidiana, e esse ambiente imp\u00f5e altas demandas \u00e0 produ\u00e7\u00e3o e \u00e0 caracteriza\u00e7\u00e3o de proteases. Portanto, a compreens\u00e3o aprofundada do mecanismo catal\u00edtico da protease, a rela\u00e7\u00e3o entre a estrutura espacial e a fun\u00e7\u00e3o catal\u00edtica e a modifica\u00e7\u00e3o direcional das cepas produtoras de enzimas para atender aos requisitos da produ\u00e7\u00e3o industrial ainda s\u00e3o a tend\u00eancia de desenvolvimento da engenharia de enzimas no futuro. Al\u00e9m disso, al\u00e9m da sele\u00e7\u00e3o e da cria\u00e7\u00e3o de boas linhagens, devem ser feitos esfor\u00e7os para tentar construir novos sistemas de express\u00e3o ou sistemas de coexpress\u00e3o de v\u00e1rios genes de protease, para construir mais vetores de express\u00e3o para proteases, de modo a realizar a express\u00e3o eficaz de diferentes componentes e melhorar ainda mais a quantidade de express\u00e3o, a atividade e a estabilidade da enzima.<\/p>\n

Entre em contato conosco agora!<\/b><\/strong><\/h2>\n

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\u00a0info@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>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Composto Glucoamilase<\/span><\/a><\/td>\n9032-08-0<\/td>\n<\/tr>\n
Pullulanase<\/span><\/a><\/td>\n9075-68-7<\/td>\n<\/tr>\n
Xilanase<\/span><\/a><\/td>\n37278-89-0<\/td>\n<\/tr>\n
Celulase<\/span><\/a><\/td>\n9012-54-8<\/td>\n<\/tr>\n
Naringinase<\/span><\/a><\/td>\n9068-31-9<\/td>\n<\/tr>\n
\u03b2-Amilase<\/span><\/a><\/td>\n9000-91-3<\/td>\n<\/tr>\n
Glucose oxidase<\/span><\/a><\/td>\n9001-37-0<\/td>\n<\/tr>\n
alfa-Amilase<\/td>\n9000-90-2<\/td>\n<\/tr>\n
Pectinase<\/span><\/a><\/td>\n9032-75-1<\/td>\n<\/tr>\n
Peroxidase<\/td>\n9003-99-0<\/td>\n<\/tr>\n
Lipase<\/span><\/a><\/td>\n9001-62-1<\/td>\n<\/tr>\n
Catalase<\/span><\/a><\/td>\n9001-05-2<\/td>\n<\/tr>\n
TANNASE<\/span><\/a><\/td>\n9025-71-2<\/td>\n<\/tr>\n
Elastase<\/span><\/a><\/td>\n39445-21-1<\/td>\n<\/tr>\n
Urease<\/span><\/a><\/td>\n9002-13-5<\/td>\n<\/tr>\n
DEXTRANASE<\/span><\/a><\/td>\n9025-70-1<\/td>\n<\/tr>\n
L-L\u00e1ctico desidrogenase<\/span><\/a><\/td>\n9001-60-9<\/td>\n<\/tr>\n
Malato desidrogenase<\/span><\/a><\/td>\n9001-64-3<\/td>\n<\/tr>\n
Colesterol oxidase<\/span><\/a><\/td>\n9028-76-6<\/td>\n<\/tr>\n<\/tbody>\n<\/table>","protected":false},"excerpt":{"rendered":"

Modifica\u00e7\u00e3o de cepas produtoras de protease A protease \u00e9 uma classe de enzima que catalisa a hidr\u00f3lise de liga\u00e7\u00f5es pept\u00eddicas em prote\u00ednas, amplamente encontrada em animais, plantas e microrganismos, e \u00e9 uma das prepara\u00e7\u00f5es enzim\u00e1ticas mais usadas em esp\u00e9cies de enzimas industriais, representando cerca de 60% da quantidade total de enzimas. Fontes microbianas de [...]<\/p>","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[108],"tags":[],"class_list":["post-7270","post","type-post","status-publish","format-standard","hentry","category-enzyme-news"],"yoast_head":"\nModification of protease-producing strains - 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\/modification-of-protease-producing-strains\/\" \/>\n<meta property=\"og:locale\" content=\"pt_BR\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Modification of protease-producing strains - Longchang Chemical\" \/>\n<meta property=\"og:description\" content=\"Modification of protease-producing strains Protease is a class of enzyme that catalyzes the hydrolysis of peptide bonds in proteins, which is widely found in animals, plants and microorganisms, and it is one of the most used enzyme preparations in industrial enzyme species, accounting for about 60% of the total amount of enzymes. 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