کد مقاله کد نشریه سال انتشار مقاله انگلیسی ترجمه فارسی نسخه تمام متن
16752 42609 2016 6 صفحه PDF ندارد دانلود رایگان
عنوان انگلیسی مقاله
Synthesis of substrates for periodate-coupled assay of phospholipases C and sphingomyelinases
ترجمه فارسی عنوان
سنتز بسترهای مورد استفاده برای بررسی پروتئین همراه با فسفولیپاز C و اسپینگزومیلیناز
کلمات کلیدی
تست مزمن پیوند؛ فسفولیپاز C (PLC)؛ اسپینومیلیناز؛ بستر روش مصنوعی. سینتیک آنزیم
Periodate-coupled assay; Phospholipase C (PLC); Sphingomyelinase; Synthetic assay substrate; Enzyme kinetics
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
چکیده انگلیسی


• Four substrate analogues of glycerophospholipids are synthesized.
• One substrate is evaluated in periodate-coupled assays of fungal sphingomyelinases.
• Periodate-containing stop solutions are optimized.
• A kinetic evaluation showed relatively high KM and low kcat values.

A series of 4-nitrophenyl (pNP) and 4-methylumbelliferyl (4MU) substrate analogues of phosphatidyl choline (PC) and phosphatidic acid (PA) were synthesized from 4-bromo-1-butene by ether formation, olefin epoxidation and ring opening with the phosphate head group. The pNP PC analogue, 4-(4-nitrophenoxy)-2-hydroxy-butyl-1-phosphoryl choline (1) was evaluated in assays of fungal sphingomyelinases, also displaying phospholipase C activity. Reactions were terminated with a periodate-containing stop solution, leading to liberation of pNP, quantified spectrophotometrically in an end-point measurement. A kinetic evaluation of sphingomyelinases from Kionochaeta sp. and Penicillium emersonii showed relatively high KM and low kcat values for this substrate, limiting its practical applicability in assays with low sphingomyelinase concentrations.

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Enzyme and Microbial Technology - Volume 91, September 2016, Pages 66–71
نویسندگان
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