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SU1217879A1 - Method of estimating activity of oxidase-l-amino acids - Google Patents

Method of estimating activity of oxidase-l-amino acids Download PDF

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Publication number
SU1217879A1
SU1217879A1 SU833642649A SU3642649A SU1217879A1 SU 1217879 A1 SU1217879 A1 SU 1217879A1 SU 833642649 A SU833642649 A SU 833642649A SU 3642649 A SU3642649 A SU 3642649A SU 1217879 A1 SU1217879 A1 SU 1217879A1
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SU
USSR - Soviet Union
Prior art keywords
activity
oxidase
amino acids
solution
optical density
Prior art date
Application number
SU833642649A
Other languages
Russian (ru)
Inventor
Темирболат Темболатович Березов
Елена Васильевна Лукашева
Ирина Павловна Смирнова
Султан Хамидович Хадуев
Original Assignee
Университет дружбы народов им.Патриса Лумумбы
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Application filed by Университет дружбы народов им.Патриса Лумумбы filed Critical Университет дружбы народов им.Патриса Лумумбы
Priority to SU833642649A priority Critical patent/SU1217879A1/en
Application granted granted Critical
Publication of SU1217879A1 publication Critical patent/SU1217879A1/en

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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Description

1one

Изобретение относитс  к биохимии, а именно к биохимическим способам определени  активности ферментов, в частности оксидазы-Ь-аминокислот.The invention relates to biochemistry, in particular to biochemical methods for determining the activity of enzymes, in particular b-amino acid oxidases.

Цель изобретени  .- сокращение времени определени  активности.The purpose of the invention. - Reducing the time to determine the activity.

При м е р. Дл  определени  активности вз ли раствор фермента, выделенного из Trichoderma sp., после частичной очистки фракционированным осаждением сульфатом аммони , К разбавили его в 100 раз. Определение ферментативной активности осу- ществл ли в кювете спектрофотометра . Реакционна  смесь содержала, об.%:An example. To determine the activity, a solution of the enzyme isolated from Trichoderma sp. Was taken, after partial purification by fractionated precipitation with ammonium sulfate, K was diluted 100 times. Determination of the enzymatic activity was performed in a cell of the spectrophotometer. The reaction mixture contained,%:

Натрий-фосфатный буфер 0,1 М, рН 7,576Sodium phosphate buffer 0.1 M, pH 7.576

L-лизин 0,1 И4L-lysine 0.1 I4

Пероксидаза 0,04% 1 Ферроцианид кали  0,1 М 4 Раствор оксндазы-Ь-амино- кислот .15Peroxidase 0.04% 1 Potassium ferrocyanide 0.1 M 4 Oxdase-L-amino acid solution .15

Реакцию иницируют добавлением раствора фермента.The reaction is initiated by the addition of an enzyme solution.

Смесь инкубировали при комнатной температуре 5 мин,после чего добав9The mixture was incubated at room temperature for 5 minutes, after which the addition9

л ли 25%-ную Тр ихлоруксусную кислоту до концентрации 3%. Измер ли оптическую плотность раствора при 420 нм (D(,0,21). В кювету контрол  вместо раствора L-лизина внесли буферный раствор. Через 5 мин оптическа  плотность в кювете контрол  составл ла Dej, 0,022. Нашли разность оптических плотностейl 25% Tp chloroacetic acid to a concentration of 3%. The optical density of the solution was measured at 420 nm (D (, 0.21). A buffer solution was added to the control instead of the L-lysine solution. After 5 min, the optical density in the control cuvette was Dej, 0.022. The optical density difference was found

D DpaB-D«fP 0,21-0,,l88.D DpaB-D «fP 0.21-0, l88.

За единицу активности фермента , (Е) прин то такое его количество, которое катализирует образование 1 мкМ перекиси водорода в минуту приPer unit of enzyme activity, (E) the amount that catalyzes the formation of 1 μM of hydrogen peroxide per minute is taken at

25 С. Калибровочную кривую построили , принима  оптическую плотность окрашенного продукта - феррицианида кали , равной 1040 М .25 C. A calibration curve was constructed taking the optical density of a colored product — potassium ferricyanide, equal to 1040 M.

По калибровочной кривой определили , что концентраци  фермента в реакционной смеси составл ет 0,038 Е/мл. Учитыва  степень разбавлени , вычислили , что ферментативна  активностьFrom the calibration curve, it was determined that the concentration of the enzyme in the reaction mixture was 0.038 U / ml. Taking into account the degree of dilution, we calculated that the enzymatic activity

раствора после осаждени  сульфатом «аммони  и диализа составл ет 0,038 « «6,,5 Е/мл.the solution after precipitation with ammonium sulfate and dialysis is 0.038 "" 6, 5 U / ml.

Claims (1)

СПОСОБ ОПРЕДЕЛЕНИЯ АКТИВНОСТИ ОКСИДАЗЫ-L-АМИНОКИСЛОТ путем инкубации’фермента в среде, содержащей фосфатный буфер, L-лизин, пероксидазу и субстрат пероксидазы с последующим измерением оптической плотности, отличающийся тем, что, с целью сокращения времени определения активности, инкубацию проводят при pH 7,5, а в качестве субстрата пероксидазы используют ферроцианид калия.METHOD FOR DETERMINING OXIDASE-L-AMINO ACIDS ACTIVITY by incubating the enzyme in a medium containing phosphate buffer, L-lysine, peroxidase and peroxidase substrate with subsequent measurement of optical density, characterized in that, in order to reduce the time for determining activity, the incubation is carried out at pH 7 , 5, and potassium ferrocyanide is used as a peroxidase substrate.
SU833642649A 1983-06-03 1983-06-03 Method of estimating activity of oxidase-l-amino acids SU1217879A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
SU833642649A SU1217879A1 (en) 1983-06-03 1983-06-03 Method of estimating activity of oxidase-l-amino acids

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
SU833642649A SU1217879A1 (en) 1983-06-03 1983-06-03 Method of estimating activity of oxidase-l-amino acids

Publications (1)

Publication Number Publication Date
SU1217879A1 true SU1217879A1 (en) 1986-03-15

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Family Applications (1)

Application Number Title Priority Date Filing Date
SU833642649A SU1217879A1 (en) 1983-06-03 1983-06-03 Method of estimating activity of oxidase-l-amino acids

Country Status (1)

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SU (1) SU1217879A1 (en)

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Авторское свидетельство СССР № 1163268, 21.04.83. *

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