Controlled Aggregation and Increased Stability of β-Glucuronidase by Cellulose Binding Domain FusionReportar como inadecuado




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Cellulose-binding domains CBDs are protein domains with cellulose-binding activity, and some act as leaders in the localization of cellulosomal scaffoldin proteins to the hydrophobic surface of crystalline cellulose. In this study, we found that a CBD fusion enhanced and improved soluble β-glucuronidase GusA enzyme properties through the formation of an artificially oligomeric state. First, a soluble CBD fused to the C-terminus of GusA GusA-CBD was obtained and characterized. Interestingly, the soluble GusA-CBD showed maximum activity at higher temperatures 65°C and more acidic pH values pH 6.0 than free GusA did 60°C and pH 7.5. Moreover, the GusA-CBD enzyme showed higher thermal and pH stabilities than the free GusA enzyme did. Additionally, GusA-CBD showed higher enzymatic activity in the presence of methanol than free GusA did. Evaluation of the protease accessibility of both enzymes revealed that GusA-CBD retained 100% of its activity after 1 h incubation in 0.5 mg-ml protease K, while free GusA completely lost its activity. Simple fusion of CBD as a single domain may be useful for tunable enzyme states to improve enzyme stability in industrial applications.



Autor: Soo-Jin Yeom , Gui Hwan Han , Moonjung Kim, Kil Koang Kwon, Yaoyao Fu, Haseong Kim, Hyewon Lee, Dae-Hee Lee, Heungchae Jung, Seun

Fuente: http://plos.srce.hr/



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