کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10537165 | 962685 | 2013 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Expression and characterization of PhzE from P. aeruginosa PAO1: aminodeoxyisochorismate synthase involved in pyocyanin and phenazine-1-carboxylate production
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کلمات کلیدی
GDHADCSADIC6-Diazo-5-oxo-l-norleucine - 6-دیازو-5-اکسو-ال-نللیسینDON - دونIsochorismate synthase - سنتاز IsochorismateAnthranilate synthase - سنتاز آنترونیلاتPseudomonas aeruginosa - سودوموناس آئروژینوزاlactate dehydrogenase - لاکتات دهیدروژناز LDH - لاکتات دهیدروژناز به صورت مختصر شده LDH Chorismate mutase - موتاز کوریسماتPyocyanin - پیوسیانینglutamate dehydrogenase - گلوتامات دهیدروژناز
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
PhzE from Pseudomonas aeruginosa catalyzes the first step in the biosynthesis of phenazine-1-carboxylic acid, pyocyanin, and other phenazines, which are virulence factors for Pseudomonas species. The reaction catalyzed converts chorismate into aminodeoxyisochorismate using ammonia supplied by a glutamine amidotransferase domain. It has structural and sequence homology to other chorismate-utilizing enzymes such as anthranilate synthase, isochorismate synthase, aminodeoxychorismate synthase, and salicylate synthase. Like these enzymes, it is Mg2 + dependent and catalyzes a similar SN2" nucleophilic substitution reaction. PhzE catalyzes the addition of ammonia to C2 of chorismate, as does anthranilate synthase, yet unlike anthranilate synthase it does not catalyze elimination of pyruvate from enzyme-bound aminodeoxyisochorismate. Herein, the cloning of the phzE gene, high level expression of active enzyme in E. coli, purification, and kinetic characterization of the enzyme is presented, including temperature and pH dependence. Steady-state kinetics give Kchorismate = 20 ± 4 μM, KMg2 + = 294 ± 22 μM, KL-gln = 11 ± 1 mM, and kcat = 2.2 ± 0.2 sâ 1 for a random kinetic mechanism. PhzE can use NH4+ as an alternative nucleophile, while Co2 + and Mn2 + are alternative divalent metals.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1834, Issue 1, January 2013, Pages 240-246
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1834, Issue 1, January 2013, Pages 240-246
نویسندگان
Justin E. Culbertson, Michael D. Toney,