Estimation of cellobiohydrolase I activity by numerical differentiation of dynamic ultraviolet spectroscopy.

نویسندگان

  • Bin Wu
  • Yue Zhao
  • Pei-Ji Gao
چکیده

1,4-beta-D-glucan cellobiohydrolase I (CBH I), p-nitrophenyl beta-D-cellobioside, p-nitrophenol and cellobiose show distinct ultraviolet spectra, allowing the design of an assay to track the dynamic process of p-nitrophenyl beta-D-cellobioside hydrolysis by CBH I. Based on the linear relationship between p-nitrophenol formation in the hydrolysate and its first derivative absorption curve of AUC340-400 nm (area under the curve), a new sensitive assay for the determination of CBH I activity was developed. The dynamic parameters of catalysis reaction, such as Vm and kcat, can all be derived from this result. The influence of beta-glucosidase and endoglucanase in crude enzyme sample on the assay was discussed in detail. This approach is useful for accurate determination of the activity of CBHs.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Photocatalyst Ag@N/TiO2 Nanoparticles: Fabrication, Characterization, and Investigation of the Effect of Coating on Methyl Orange Dye Degradation

In this research, N-doped TiO2 (N/TiO2) nanoparticles have been synthesized by a sol-gel method. N/TiO2 nanoparticle has been coated with Ag metal by photochemical method. Triethylamine, N(CH3CH2)3, have been used as precursors of Nitrogen, titanium tetraiso-propoxide (TTIP), Ti[OCH(CH3)2]4, used as precursors of titanium and Ag(NO3)2 used as precursors of Silver in synthesis of these nanoparti...

متن کامل

Optimal Wavelength Selection in Ultraviolet Spectroscopy for the Estimation of Toxin Reduction Ratio during Hemodialysis

Introduction The concentration of substances, including urea, creatinine, and uric acid, can be used as an index to measure toxic uremic solutes in the blood during dialysis and interdialytic intervals. The on-line monitoring of toxin concentration allows for the clearance measurement of some low-molecular-weight solutes at any time during hemodialysis.The aim of this study was to determine the...

متن کامل

Manganese mine actinobacterial mediated gold nanoparticles synthesis and their antibacterial activity

Background and Objectives: Actinobacteria efficiently can produce different nanoparticles with different biological properties due to their ability to produce secondary metabolites. The aim of present study, isolation and screening of gold nanoparticles via producing actinobacteria from the soil were studied and their antibacterial activities was evaluate. Methods: In this study, after iso...

متن کامل

Green Biosynthesis of Silver Nanoparticles Using Glycyrrhiza glabra L. Extract and Evaluation of their Selective Antimicrobial Activity

The aim of present study is to determine the content of some phytochemicals in stem and leaf extracts of the medicinal plant of Glycyrrhiza glabra and design a green method for biosynthesis of silver nanoparticles by these extracts as stabilizers and bioreducers. The properties of the synthesized silver nanoparticles (AgNPs) were monitored by using Ultraviolet-Visible absorption spectroscopy (U...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Acta biochimica et biophysica Sinica

دوره 38 6  شماره 

صفحات  -

تاریخ انتشار 2006