{"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\/tr\/modification-of-protease-producing-strains\/","title":{"rendered":"Proteaz \u00fcreten su\u015flar\u0131n modifikasyonu"},"content":{"rendered":"

Proteaz \u00fcreten su\u015flar\u0131n modifikasyonu<\/strong><\/h1>\n

Proteaz, proteinlerdeki peptit ba\u011flar\u0131n\u0131n hidrolizini katalize eden, hayvanlarda, bitkilerde ve mikroorganizmalarda yayg\u0131n olarak bulunan bir enzim s\u0131n\u0131f\u0131d\u0131r ve end\u00fcstriyel enzim t\u00fcrlerinde en \u00e7ok kullan\u0131lan enzim preparatlar\u0131ndan biridir ve toplam enzim miktar\u0131n\u0131n yakla\u015f\u0131k 60%'sini olu\u015fturur. Mikrobiyal proteaz kaynaklar\u0131 bol, \u00e7e\u015fitli ve \u00fcretimi kolayd\u0131r ve piyasadaki proteazlar\u0131n ana kayna\u011f\u0131 haline gelir. Proteaz\u0131n ticari \u00fcretimi ge\u00e7en y\u00fczy\u0131l\u0131n ba\u015flar\u0131na kadar uzanmaktad\u0131r. 1908 y\u0131l\u0131nda Almanlar deri tabaklamada pankreatik enzim kullanmaya ba\u015flam\u0131\u015f, 1911 y\u0131l\u0131nda Amerika Birle\u015fik Devletleri Wallerstein \u015firketi bira i\u00e7in berrakla\u015ft\u0131r\u0131c\u0131 bir madde olarak papain \u00fcretmi\u015ftir. 60'l\u0131 y\u0131llar\u0131n ba\u015f\u0131nda Hollanda proteaz ilaveli deterjanlar \u00fcretmi\u015ftir. Buna ek olarak, \u0131s\u0131ya dayan\u0131kl\u0131, aside dayan\u0131kl\u0131, tuza dayan\u0131kl\u0131 proteaz \u00fcretimi bakteri se\u00e7imi ve \u0131slah\u0131n\u0131n yan\u0131 s\u0131ra alkali proteaz ara\u015ft\u0131rmalar\u0131n\u0131n hammadde fermantasyon \u00fcretimi olarak petrol\u00fcn ger\u00e7ekle\u015ftirilmesi i\u00e7in pratik ile birle\u015ftirilmi\u015ftir.<\/p>\n

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Bir\u00e7ok proteaz t\u00fcr\u00fc vard\u0131r ve molek\u00fcler a\u011f\u0131rl\u0131k boyutlar\u0131, uzaysal yap\u0131lar\u0131 ve i\u015flevleri \u00e7ok farkl\u0131d\u0131r, ancak her proteaz ailesi hala olduk\u00e7a korunmu\u015f bir yap\u0131sal alan\u0131 korumaktad\u0131r. \u015eu anda, 100'den fazla proteaz t\u00fcr\u00fc kristalize edilmi\u015f veya y\u00fcksek oranda safla\u015ft\u0131r\u0131lm\u0131\u015f ve bir\u00e7o\u011fu birincil ve stereo yap\u0131lar\u0131 (tersiyer yap\u0131) a\u00e7\u0131s\u0131ndan ayd\u0131nlat\u0131lm\u0131\u015ft\u0131r.<\/p>\n

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2 Proteaz su\u015flar\u0131n\u0131n modifikasyonu<\/p>\n

Proteaz\u0131n verimli bir \u015fekilde ifade edilmesi ve enzimatik \u00f6zelliklerinin optimizasyonu i\u00e7in m\u00fchendislik bakterilerinin yap\u0131m\u0131 yerli ve yabanc\u0131 akademisyenlerin ilgi oda\u011f\u0131 olmu\u015ftur ve genellikle kullan\u0131lan ara\u00e7lar \u015funlard\u0131r: mutasyon \u0131slah\u0131, protoplazmik f\u00fczyon, geneti\u011fi de\u011fi\u015ftirilmi\u015f bakterilerin yap\u0131m\u0131 ve in vitro y\u00f6nl\u00fc evrim teknolojisi.<\/p>\n

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2.1 Mutajenez \u0131slah\u0131 ve protoplazmik f\u00fczyon<\/p>\n

Mutajenez \u0131slah\u0131, m\u00fckemmel \u00f6zelliklere sahip mutant su\u015flar elde etmek i\u00e7in bakterinin genetik materyalinde mutasyonlar\u0131 ind\u00fcklemek i\u00e7in esas olarak radyasyon mutajenezi veya baz\u0131 kimyasal reaktifler yoluyla yap\u0131l\u0131r. \u00d6rne\u011fin, Cai Wanling ve arkada\u015flar\u0131 taraf\u0131ndan enzim \u00fcreten su\u015flar\u0131n UV mutagenezinden sonra, enzim aktiviteleri orijinal su\u015fa k\u0131yasla 16% artm\u0131\u015ft\u0131r. Yanli Li ve arkada\u015flar\u0131, Aspergillus oryzae ZW-06'n\u0131n n\u00f6tr proteaz \u00fcreten enzim aktivitesini Co60-y\u00f6nlendirmeli mutagenez ile 15.000 U\/g kuru lor'a kadar taram\u0131\u015f ve bu de\u011fer mutagenez \u00f6ncesine g\u00f6re 74% daha y\u00fcksek olmu\u015ftur. Huang Hongying ve ark. d\u00fc\u015f\u00fck enerjili N+ iyon implantasyon teknolojisini vah\u015fi tip Bacillus licheniformis \u00fczerinde d\u00f6rt kez farkl\u0131 fizikokimyasal mutajenez y\u00f6ntemleriyle birle\u015ftirerek y\u00fcksek verimli bir su\u015f elde etmi\u015flerdir; ham enzim aktivitesi 12425. 9 U\/mL, \u00e7\u0131k\u0131\u015f su\u015fundan 17,1 kat daha y\u00fcksektir.<\/p>\n

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Protoplazmik f\u00fczyon teknolojisi, iki ebeveynli bir su\u015fun h\u00fccrelerinin protoplazmik f\u00fczyon i\u00e7in duvarlar\u0131n\u0131n kald\u0131r\u0131ld\u0131\u011f\u0131, b\u00f6ylece stabil rekombinantlar elde etmek i\u00e7in genomlar\u0131n\u0131n de\u011fi\u015ftirildi\u011fi ve rekombine edildi\u011fi bir s\u00fcre\u00e7tir. \u00d6rne\u011fin, Pan Yanyun ve arkada\u015flar\u0131, Bacillus licheniformis 2709'u alkalin proteaz gen klonlama vekt\u00f6r\u00fc pDW2 i\u00e7eren Bacillus subtilis BD105 ile kayna\u015ft\u0131rmak i\u00e7in protoplazmik f\u00fczyon y\u00f6ntemini kullanarak, 2709'dan 50%-100% daha y\u00fcksek fermantasyon enzimi \u00fcretimi ve sallama \u015fi\u015fesi testinde 30.000 U\/mL'ye kadar enzim \u00fcretimi ile y\u00fcksek verimli bir A16 su\u015fu elde etmi\u015flerdir.<\/p>\n

 <\/p>\n

2.2 Genetik m\u00fchendisli\u011fi bakterilerinin yap\u0131m\u0131<\/p>\n

M\u00fchendislik bakterilerinin yap\u0131m\u0131 genellikle uygun ifade konak\u00e7\u0131lar\u0131 ve ifade vekt\u00f6rleri veya ifade elemanlar\u0131 gerektirir. \u0130lk olarak, hedef gen rekombinant vekt\u00f6r elde etmek i\u00e7in ekspresyon vekt\u00f6r\u00fcne klonlan\u0131r, daha sonra tarama s\u00fcreci boyunca ekspresyon kona\u011f\u0131na d\u00f6n\u00fc\u015ft\u00fcr\u00fcl\u00fcr ve son olarak y\u00fcksek verimli su\u015f s\u00fcreci elde edilir. Y\u00fcksek verimli su\u015flar elde etmek i\u00e7in eksojen hedef gen par\u00e7alar\u0131n\u0131 do\u011frudan kona\u011f\u0131n genomuna entegre etmek de m\u00fcmk\u00fcnd\u00fcr.<\/p>\n

 <\/p>\n

[18-20 Ekim] 2024 2. \u0130leri Enzim M\u00fchendisli\u011fi ve Enzim Teknolojisi Uygulamalar\u0131 Konferans\u0131<\/p>\n

Min Zhang ve arkada\u015flar\u0131 sacB geninin promot\u00f6r-sinyal peptit dizisini (sacR) Bacillus subtilis n\u00f6tral proteaz geni ile ba\u011flayarak pHP13 vekt\u00f6r\u00fcne klonlam\u0131\u015f, n\u00f6tral proteaz genini i\u00e7eren ind\u00fcklenebilir ekspresyon salg\u0131lama vekt\u00f6r\u00fc pHP13SN'yi olu\u015fturmu\u015f ve ard\u0131ndan bunu Bacillus subtilis DB104'e d\u00f6n\u00fc\u015ft\u00fcrm\u00fc\u015ft\u00fcr; bu da \u00e7\u0131k\u0131\u015f su\u015funa k\u0131yasla enzim canl\u0131l\u0131\u011f\u0131nda 48% art\u0131\u015f g\u00f6stermi\u015ftir. WangHui ve arkada\u015flar\u0131 Bacillus amyloliquefaciens K11'den n\u00f6tral bir proteaz olan Banpr'\u0131 klonlam\u0131\u015f ve ifade kona\u011f\u0131 olarak Bacillus amyloliquefaciens K11 ile rekombinant bir ifade plazmidi, pUB110-Banpr, olu\u015fturmu\u015f ve sallama \u015fi\u015fesi fermantasyonunda enzim aktivitesi 8995 \u00b1 250 U\/mL'ye ve 15 L fermantasyon sisteminde 28084 \u00b1 1282 U\/mL'ye ula\u015fm\u0131\u015ft\u0131r ki bu end\u00fcstriyel Bacillus subtilis AS su\u015fundan \u00e7ok daha fazlad\u0131r.1398 (8000-10000 U\/mL).<\/p>\n

 <\/p>\n

2.3 Proteaz \u00f6zelliklerinin iyile\u015ftirilmesi<\/p>\n

\u0130n vitro y\u00f6nlendirilmi\u015f evrim tekniklerinin geli\u015fmesiyle birlikte, hedef proteinin \u00fc\u00e7 boyutlu yap\u0131sal bilgisi ve etki mekanizmas\u0131 bilinmeden kodlay\u0131c\u0131 genlerin rastgele mutasyonu, DNAShuffling teknikleri ve y\u00f6nlendirilmi\u015f tarama ile geli\u015ftirilmi\u015f veya yeni i\u015flevlere sahip proteinler elde edilebilmektedir.<\/p>\n

 <\/p>\n

 <\/p>\n

Enzim stabilitesi, ticarile\u015ftirmenin ba\u015far\u0131s\u0131nda kilit bir fakt\u00f6rd\u00fcr. Enzim stabilitesi, tuz k\u00f6pr\u00fcleri, dis\u00fclfit ba\u011flar\u0131, aromatik etkile\u015fimler ve enzimin kalsiyum iyonlar\u0131na olan afinitesinin art\u0131r\u0131lmas\u0131yla geli\u015ftirilebilir ve b\u00f6ylece hedeflenen mutagenez yoluyla molek\u00fcl\u00fcn sertli\u011fi art\u0131r\u0131labilir. BPN' proteaz\u0131n\u0131n Gly61 veya Ser98'inin Cys ile veya her ikisinin birden de\u011fi\u015ftirilmesi, katalitik etkinlikte bir de\u011fi\u015fiklik olmaks\u0131z\u0131n enzimin termal stabilitesinde bir art\u0131\u015fla sonu\u00e7lanm\u0131\u015ft\u0131r.<\/p>\n

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B. subtilis proteaz\u0131n\u0131n Val72'sinin Ile, Ala92'sinin Thr ve Gly131'inin Asp ile de\u011fi\u015ftirilmesiyle, mutant enzim 10\u00b0C'de vah\u015fi enzimden 2 kat daha y\u00fcksek substrat aktivitesini katalize edebilir. 2005 y\u0131l\u0131nda Danimarkal\u0131 Novozymes \u015firketi, deterjana eklenebilen d\u00fc\u015f\u00fck s\u0131cakl\u0131k proteaz\u0131 \"Polarzyme \"\u0131 tan\u0131tt\u0131. Deterjana eklenmesiyle y\u0131kama s\u0131cakl\u0131\u011f\u0131 60\u00b0C ve 40\u00b0C'den 30\u00b0C ve 20\u00b0C'ye d\u00fc\u015fm\u00fc\u015f, 60% elektrik enerjisi tasarrufu sa\u011flanm\u0131\u015f ve d\u00fc\u015f\u00fck s\u0131cakl\u0131kta proteaz i\u00e7eren \"carecoldwash\" deterjan\u0131n\u0131n sat\u0131\u015f hacmi yedi kat artm\u0131\u015ft\u0131r.<\/p>\n

 <\/p>\n

3 G\u00f6r\u00fcn\u00fcm<\/p>\n

Proteazlar g\u00fcnl\u00fck hayat\u0131m\u0131z\u0131 yak\u0131ndan ilgilendiren \u00e7e\u015fitli alanlarda kullan\u0131lmaktad\u0131r ve bu ortam proteazlar\u0131n \u00fcretimi ve karakterizasyonu konusunda y\u00fcksek talepler do\u011furmaktad\u0131r. Bu nedenle, proteaz\u0131n katalitik mekanizmas\u0131n\u0131n derinlemesine anla\u015f\u0131lmas\u0131, uzamsal yap\u0131 ve katalitik fonksiyon aras\u0131ndaki ili\u015fki ve end\u00fcstriyel \u00fcretimin gereksinimlerini kar\u015f\u0131lamak i\u00e7in enzim \u00fcreten su\u015flar\u0131n y\u00f6nl\u00fc modifikasyonu, gelecekte enzim m\u00fchendisli\u011finin geli\u015fme e\u011filimidir. Buna ek olarak, iyi su\u015flar\u0131n se\u00e7imi ve \u0131slah\u0131n\u0131n yan\u0131 s\u0131ra, farkl\u0131 bile\u015fenlerin etkili ifadesini ger\u00e7ekle\u015ftirmek ve ifade miktar\u0131n\u0131, enzim aktivitesini ve enzim stabilitesini daha da iyile\u015ftirmek i\u00e7in proteazlar i\u00e7in daha fazla ifade vekt\u00f6r\u00fc olu\u015fturmak \u00fczere yeni ifade sistemleri veya \u00e7oklu proteaz geni birlikte ifade sistemleri olu\u015fturmaya \u00e7al\u0131\u015fmak i\u00e7in \u00e7aba g\u00f6sterilmelidir.<\/p>\n

\u015eimdi Bize Ula\u015f\u0131n!<\/b><\/strong><\/h2>\n

Fiyata ihtiyac\u0131n\u0131z varsa, l\u00fctfen a\u015fa\u011f\u0131daki forma ileti\u015fim bilgilerinizi doldurun, genellikle 24 saat i\u00e7inde sizinle ileti\u015fime ge\u00e7ece\u011fiz. Bana e-posta da g\u00f6nderebilirsiniz\u00a0info@longchangchemical.com<\/a><\/span>\u00a0\u00c7al\u0131\u015fma saatleri i\u00e7inde (8:30 - 6:00 UTC+8 Pzt.~Sat.) veya h\u0131zl\u0131 yan\u0131t almak i\u00e7in web sitesi canl\u0131 sohbetini kullan\u0131n.<\/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
Bile\u015fik Glukoamilaz<\/span><\/a><\/td>\n9032-08-0<\/td>\n<\/tr>\n
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Kolesterol oksidaz<\/span><\/a><\/td>\n9028-76-6<\/td>\n<\/tr>\n<\/tbody>\n<\/table>","protected":false},"excerpt":{"rendered":"

Proteaz \u00fcreten su\u015flar\u0131n modifikasyonu Proteaz, hayvanlarda, bitkilerde ve mikroorganizmalarda yayg\u0131n olarak bulunan proteinlerdeki peptit ba\u011flar\u0131n\u0131n hidrolizini katalize eden bir enzim s\u0131n\u0131f\u0131d\u0131r ve end\u00fcstriyel enzim t\u00fcrlerinde en \u00e7ok kullan\u0131lan enzim preparatlar\u0131ndan biridir ve toplam enzim miktar\u0131n\u0131n yakla\u015f\u0131k 60%'sini olu\u015fturur. Mikrobiyal kaynaklar\u0131 [...]<\/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\/tr\/modification-of-protease-producing-strains\/\" \/>\n<meta property=\"og:locale\" content=\"tr_TR\" \/>\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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