Preparation and Kinetic Properties of a New Form of Chymotrypsin Which Is Active at Alkaline pH

نویسندگان

  • PABLO VALENZUELA
  • MYRON L. BENDER
چکیده

The preparation of a new stable and active form of chymotrypsin is described. The enzyme possesses threonine-147 instead of alanineas the NH&erminal group of the C chain and has been called ai-chymotrypsin. The conformational transition that affects chymotrypsins at alkaline pH was investigated and compared with that of 01and &chymotrypsins by studying the kinetic constants and their pH dependencies for the hydrolysis of various specific ester substrates. The keal values obtained with LYEchymotrypsin are similar to those of o(and 6-chymotrypsins. The K, values showed a progressive increase toward the alkaline pH region. The shape of the K,-pH profiles closely resemble those of &chym.otrypsin and differ considerably from the behavior of cr-chymotrypsin. The results strongly implicate the participation of the alanineamino group in the reversible inactivation of oc-chymotrypsin at high pH.

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

Preparation and Kinetic Properties of a New Form of Chymotrypsin Which Is Active at Alkaline pH : a,-Chymotrypsin*

The preparation of a new stable and active form of chymotrypsin is described. The enzyme possesses threonine-147 instead of alanineas the NH&erminal group of the C chain and has been called ai-chymotrypsin. The conformational transition that affects chymotrypsins at alkaline pH was investigated and compared with that of 01and &chymotrypsins by studying the kinetic constants and their pH depende...

متن کامل

Preparation and kinetic properties of a new form of chymotrypsin which is active at alkaline pH: 1 -chymotrypsin.

The preparation of a new stable and active form of chymotrypsin is described. The enzyme possesses threonine-147 instead of alanineas the NH&erminal group of the C chain and has been called ai-chymotrypsin. The conformational transition that affects chymotrypsins at alkaline pH was investigated and compared with that of 01and &chymotrypsins by studying the kinetic constants and their pH depende...

متن کامل

Investigations of the chymotrypsin-catalyzed hydrolysis of specific substrates. I. The pH dependence of the catalytic hydrolysis of N-acetyl-L-tryptophanamide by three forms of the enzyme at alkaline pH.

The pH dependence of the steady state kinetic parameters K,(app) and $,t of the a-, acetylated 6-, and ar-chymotrypsincatalyzed hydrolysis of the specific amide substrate N-acetylr.-tryptophanamide, has been investigated in the neutral and alkaline pH regions. A completely automatic technique for measuring the hydrolysis of amides was used. New results and important aspects of the chymotrypsin-...

متن کامل

Fluorotyrosine alkaline phosphatase from Escherichia coli: preparation, properties, and fluorine-19 nuclear magnetic resonance spectrum.

Alkaline phosphatase (EC 3.1.3.1) containing m-flurotyrosine has been prepared from E. coli grown in the presence of m-flurotyrosine. The kinetic properties of the m-fluorotyrosine enzyme measured with p-nitrophenylphosphate at pH 8.0 and dinitrophenylphosphate at pH 5.5 are essentially the same as those of normal alkaline phosphatase. However, the ability of the m-fluorotyrosine protein to ref...

متن کامل

Effect of Sodium Dodecyl Sulphate on Partial Purified Polyphenol Oxidase Activity in Red and Green Tomatoes (Solanum Lycopersicum)

In order to better understand how to prevent enzymatic greenning, it is important to understand  kinetic properties of polyphenol oxidase.  we investigated the effect of SDS on the rate of catechol oxidation by small cherry tomato partial purified PPO. PPO activity increased with increasing SDS concentration. The most effective concentration of SDS was 0.8, 1 and 1.25 mM in according to pH and ...

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:

دوره   شماره 

صفحات  -

تاریخ انتشار 2002