Indoxyl Acetate as a Substrate for Analysis of Lipase Activity

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60162694%3AG44__%2F19%3A00555343" target="_blank" >RIV/60162694:G44__/19:00555343 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11160/19:10410227

Výsledek na webu

<a href="http://downloads.hindawi.com/journals/ijac/2019/8538340.pdf" target="_blank" >http://downloads.hindawi.com/journals/ijac/2019/8538340.pdf</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1155/2019/8538340" target="_blank" >10.1155/2019/8538340</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Indoxyl Acetate as a Substrate for Analysis of Lipase Activity

Popis výsledku v původním jazyce

Lipases play a crucial role in metabolism of microbes, fungi, plants, and animals, and in analytical chemistry, they are often used in detection of fats and triglycerides. Determination of lipase activity is also important in toxicology, when lipase activity can be both increased and decreased by organophosphates and other pesticides and in medicine for diagnosis of heart diseases. The standard method for lipase activity determination is based on cleaving ester bonds in lipase buffer containing Tween. Our aim was to find a method with faster and more sensitive response. It is known that acetylcholinesterase belongs to the same group of hydrolases enzymes as lipases and it cleaves indoxyl acetate, so we assume indoxyl acetate could report a similar reaction with lipase. Our method is based on indoxyl acetate as a substrate for lipase, where indoxyl acetate is cleaved by lipase to indoxyl and acetate moiety and blue indigo is created. The method was optimized for different times and amount of enzyme and compared with the standard Tween assay. The calibration curve measured in reaction time 20 minutes with 10 mu l of lipase exhibited the best analytical parameters, and it showed Michaelis-Menten response with the Michaelis-Menten constant equal to 8.72 mmol/l. The indoxyl acetate-based method showed faster and more sensitive response than the standard method for lipase activity determination, so it has great potential in biosensor construction and it could be used in industry, medicine, toxicology, and common practice where the activity of lipases is need to be measured.

Název v anglickém jazyce

Indoxyl Acetate as a Substrate for Analysis of Lipase Activity

Popis výsledku anglicky

Lipases play a crucial role in metabolism of microbes, fungi, plants, and animals, and in analytical chemistry, they are often used in detection of fats and triglycerides. Determination of lipase activity is also important in toxicology, when lipase activity can be both increased and decreased by organophosphates and other pesticides and in medicine for diagnosis of heart diseases. The standard method for lipase activity determination is based on cleaving ester bonds in lipase buffer containing Tween. Our aim was to find a method with faster and more sensitive response. It is known that acetylcholinesterase belongs to the same group of hydrolases enzymes as lipases and it cleaves indoxyl acetate, so we assume indoxyl acetate could report a similar reaction with lipase. Our method is based on indoxyl acetate as a substrate for lipase, where indoxyl acetate is cleaved by lipase to indoxyl and acetate moiety and blue indigo is created. The method was optimized for different times and amount of enzyme and compared with the standard Tween assay. The calibration curve measured in reaction time 20 minutes with 10 mu l of lipase exhibited the best analytical parameters, and it showed Michaelis-Menten response with the Michaelis-Menten constant equal to 8.72 mmol/l. The indoxyl acetate-based method showed faster and more sensitive response than the standard method for lipase activity determination, so it has great potential in biosensor construction and it could be used in industry, medicine, toxicology, and common practice where the activity of lipases is need to be measured.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

—

Návaznosti

S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

International Journal of Analytical Chemistry

ISSN

1687-8760

e-ISSN

1687-8779

Svazek periodika

2019

Číslo periodika v rámci svazku

Dec

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

7

Strana od-do

8538340

Kód UT WoS článku

000503428200001

EID výsledku v databázi Scopus

2-s2.0-85076925556