Horseradish Peroxidase Inactivation: Heme Destruction and Influence of Polyethylene Glycol

نویسندگان

  • Liang Mao
  • Siqiang Luo
  • Qingguo Huang
  • Junhe Lu
چکیده

Horseradish peroxidase (HRP) mediates efficient conversion of many phenolic contaminants and thus has potential applications for pollution control. Such potentially important applications suffer however from the fact that the enzyme becomes quickly inactivated during phenol oxidation and polymerization. The work here provides the first experimental data of heme consumption and iron releases to support the hypothesis that HRP is inactivated by heme destruction. Product of heme destruction is identified using liquid chromatography with mass spectrometry. The heme macrocycle destruction involving deprivation of the heme iron and oxidation of the 4-vinyl group in heme occurs as a result of the reaction. We also demonstrated that heme consumption and iron releases resulting from HRP destruction are largely reduced in the presence of polyethylene glycol (PEG), providing the first evidence to indicate that heme destruction is effectively suppressed by co-dissolved PEG. These findings advance a better understanding of the mechanisms of HRP inactivation.

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

Structural insights into the effects of charge-reversal substitutions at the surface of horseradish peroxidase

Horseradish peroxidase (HRP), has gained significant interests in biotechnology, especially in biosensor field and diagnostic test kits. Hence, its solvent-exposed lysine residues 174, 232, and 241 have been frequently modified with the aim of improving its stability and catalytic efficiency. In this computational study, we investigated the effects of Lys-to-Glu substitutions on HRP structure t...

متن کامل

Catalytic Oxidation of Phenol and 2,4-Dichlorophenol by Using Horseradish Peroxidase Immobilized on Graphene Oxide/Fe₃O₄.

Graphene oxide/Fe₃O₄ (GO/Fe₃O₄) nanoparticles were synthesized by an ultrasonic-assisted reverse co-precipitation method, and then horseradish peroxidase (HRP) was covalently immobilized onto GO/Fe₃O₄ with 1-ethyl-3-(3-dimethyaminopropyl)carbodiimide (EDC) as a cross-linking agent. In order to enhance the phenol removal efficiency and prevent the inactivation of the enzyme, the polyethylene gly...

متن کامل

Enzyme immunoassay of estrogen-like substances in plasma, with polyethylene glycol as precipitant.

We devised an enzyme immunoassay for plasma estrogen-like substances in pregnancy, using estriol covalently linked to horseradish peroxidase. Free and bound steroid were separated by use of polyethylene glycol reagent. The assay satisfies the normal criteria of specificity and precision. Results obtained by radioimmunoassay and enzyme immunoassay agreed well. Our assay is particularly applicabl...

متن کامل

Elimination of 4-chlorophenol from Water Solutions Using Commercial Peroxidases

A comparative study of the two most widely used commercial peroxidases (E.C. 1.11.1.7) for removing 4-chlorophenol from aqueous industrial effluents is presented. Both the peroxidases tested, horseradish peroxidase (HRP) and soybean peroxidase (SBP), showed maximal removal efficiency in a neutral pH medium although they maintained more than 70 % of their activity in a pH range of between 6.0 an...

متن کامل

Self-assembled architectures from biohybrid triblock copolymers.

The synthesis and self-assembly behavior of biohybrid ABC triblock copolymers consisting of a synthetic diblock, polystyrene-b-polyethylene glycol (PSm-b-PEG113), where m is varied, and a hemeprotein, myoglobin (Mb) or horse radish peroxidase (HRP), is described. The synthetic diblock copolymer is first functionalized with the heme cofactor and subsequently reconstituted with the apoprotein or ...

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 3  شماره 

صفحات  -

تاریخ انتشار 2013