The Determination of Enzyme Specific Activity

Summary

This video script details an experiment on determining enzyme specific activity, focusing on glutathione S-transferase (GST). It covers the preparation of cytosolic fractions from mice, measuring GST activity using the CDNB method, creating a BSA standard curve with the Biuret method, and calculating protein content and enzyme specific activity. The script also discusses the importance of GST in detoxification and its relevance in cancer treatment, where inhibiting GST can enhance the effectiveness of chemotherapy.

Takeaways

• 🔬 The experiment aims to determine the specific activity of glutathione S-transferase (GST) enzyme and protein concentration using the Bradford method.
• 🧬 The activity of GST enzyme in protein mixtures is dependent on the amount of protein used, necessitating the knowledge of protein content in the mixture.
• 📚 Monomer and polymer concepts are introduced, with monomers being the building blocks like amino acids, and polymers being long chains like proteins.
• 🌟 Amino acids, the monomers of proteins, are differentiated by their R groups and can polymerize to form linear chains, such as peptides.
• 🔍 Three types of protein quantitative analysis are discussed: volumetric, gasometric, and spectrophotometry, with the experiment using visible light spectrophotometry.
• 📈 The Bradford method is chosen for protein quantification, which involves a color reaction with Coomassie Brilliant Blue G-250 dye, measured at 550 nm.
• 🛠️ The preparation of cytosolic fraction containing GST from mice involves several steps of centrifugation to isolate the enzyme.
• 🧪 The enzyme activity of GST is determined using the CDNB method, which measures the formation of a conjugate product at 340 nm over time.
• 📊 A standard curve using bovine serum albumin (BSA) is created to measure the concentration of the GST enzyme in the sample.
• ⚖️ The specific activity of the enzyme is calculated by dividing the enzyme units by the protein concentration, indicating the enzyme's purity and efficiency.
• 💊 The importance of measuring enzyme activities is highlighted, particularly in pharmaceutical fields, where GST enzymes play a role in detoxification and cancer treatment.

Q & A

What is the main objective of the experiment described in the script?

The main objective of the experiment is to determine the specific activity of the GST enzyme, measure the protein concentration using the Biuret method, and understand the fundamentals of enzyme activity in relation to protein content.

What is the difference between a monomer and a polymer as described in the script?

A monomer is a single small molecule that can be joined with other identical molecules to form a polymer. In the context of proteins, amino acids are the monomers, and proteins are the polymers formed by the linkage of these amino acids.

What are the elements that typically make up amino acids, the monomers of proteins?

Amino acids, which are the monomers of proteins, typically contain elements such as hydrogen (H), nitrogen (N), oxygen (O), and carbon (C).

What are the different types of protein quantitative analysis mentioned in the script?

The script mentions three types of protein quantitative analysis: volumetric analysis, gasometric analysis, and spectrophotometry, with the latter including UV and visible light spectrophotometry.

Why is it necessary to measure the protein content in a sample before determining enzyme activity?

Measuring the protein content is necessary because the activity of enzymes like GST in a protein mixture depends on the amount of protein present. This information is crucial for accurately determining the specific activity of the enzyme.

What is the Biuret method used for in the experiment?

The Biuret method is used in the experiment to determine the protein concentration in the cytosolic fraction containing the GST enzyme by measuring the color complex formed between the protein and the Biuret reagent at a wavelength of 550 nanometers.

What are the advantages and disadvantages of using the Biuret method for protein quantification?

The advantages of the Biuret method include its simplicity and speed, and the fact that the protein concentration measured is comparable to the polypeptide bond. Disadvantages include lower sensitivity and sometimes unstable color development.

What is the significance of measuring enzyme activities and protein concentrations in pharmaceutical fields?

Measuring enzyme activities and protein concentrations is significant in pharmaceutical fields because it helps in understanding the detoxification processes, such as the role of GST enzymes in the excretion of xenobiotics and chemotherapeutic agents, which can be crucial in the treatment of diseases like cancer.

What is the purpose of creating a standard curve using the BSA method in the experiment?

The purpose of creating a standard curve using the BSA (Bovine Serum Albumin) method is to measure the concentration of the GST enzyme in the cytosolic fraction by comparing the absorbance values obtained from the sample to the known concentrations of BSA.

How is the specific activity of an enzyme calculated in the experiment?

The specific activity of an enzyme is calculated by dividing the total enzyme units, which is the amount of enzyme that catalyzes one micromole of substrate reaction per minute, by the protein concentration in milligrams per milliliter.

Outlines

00:00

🔬 Introduction to Enzyme Specific Activity Experiment

The video introduces an experiment aimed at determining the specific activity of the glutathione S-transferase (GST) enzyme. It explains the importance of knowing the protein content in a sample to accurately measure enzyme activity. The video uses monomers and polymers as an analogy to explain proteins and their building blocks, amino acids. It further discusses the quantification of proteins through various methods including volumetric, gasometric, and spectrophotometry, with a focus on the visible light spectrophotometry using the Biuret method. The Biuret reaction, which measures the protein content by the color complex formed with copper ions, is highlighted for its simplicity and speed, despite being less sensitive.

05:02

🧪 Understanding GST Enzyme and Its Role in Detoxification

This section delves into the role of the GST enzyme in phase II detoxification reactions, which are crucial for the excretion of xenobiotics from the body. It explains the involvement of three different enzymes in this process, with a focus on GST. The video describes the types of substrates that GST can act upon and the different mechanisms of GST conjugation. It also discusses the various types of GST enzymes and their substrates, including CDNB, which is used in the experiment. The preparation steps for the cytosolic fraction containing GST from mice are outlined, including the use of male rats, fasting, sacrifice, and centrifugation techniques to isolate the enzyme.

10:04

📊 Measuring Enzyme Activity and Protein Content

The video script outlines the steps for measuring the GST enzyme activity using the CDNB method and preparing a standard curve using the Biuret method. It details the procedure for preparing different fractions and the importance of using distilled water as a blank. The process of mixing reagents, measuring absorbance at specific time intervals, and calculating the enzyme activity is explained. The script also covers the creation of a BSA standard curve for determining protein concentration and the steps for measuring the protein content in the cytosolic fraction containing GST.

15:06

📈 Creating a Standard Curve and Calculating Enzyme Specific Activity

This part of the script focuses on the creation of a standard curve using the Biuret method to measure the concentration of the GST enzyme. It describes the preparation of a series of BSA concentrations and the procedure for reading absorbance to plot the standard curve. The video explains how to use the standard curve to determine the protein concentration in the sample. It also discusses the calculation of enzyme specific activity, emphasizing its importance in assessing enzyme purity and understanding the efficiency of the GST enzyme in detoxification processes.

20:07

💊 Significance of GST Enzyme in Pharmaceutical Research

The final paragraph highlights the significance of the GST enzyme in pharmaceutical research, particularly in the context of cancer treatment. It discusses how GST enzymes play a role in the excretion of chemotherapeutic agents and how inhibiting these enzymes can enhance the effectiveness of cancer treatments. The video concludes by emphasizing the importance of understanding and measuring enzyme activities, especially in relation to detoxification and the development of antitumor agents.

Keywords

💡Enzyme Specific Activity

Enzyme specific activity refers to the rate at which an enzyme catalyzes a reaction under specific conditions, normalized to the amount of enzyme present. In the video, this concept is central to understanding how the activity of the glutathione S-transferase (GST) enzyme is measured. The script discusses the importance of determining this value to assess the enzyme's function and efficiency in biological processes.

💡Protein Concentration

Protein concentration is a measure of the amount of protein present in a sample, typically expressed in milligrams per milliliter. In the context of the video, determining protein concentration is essential for calculating enzyme specific activity. The script mentions using the Bradford method to quantify protein, which is a common assay in biochemistry.

💡Monomer

A monomer is a single molecule that can be joined to other identical molecules to form a polymer. In the script, monomers are described as the building blocks of proteins, such as amino acids, which are the monomers that polymerize to form polypeptide chains, ultimately creating proteins.

💡Polymer

A polymer is a large molecule composed of repeating structural units, known as monomers. In the video, polymers are related to proteins, which are long chains of amino acids. The script uses the analogy of a polymer to explain the structure of proteins and their formation from amino acid monomers.

💡Amino Acid

Amino acids are organic compounds containing an amine (-NH2) and a carboxyl (-COOH) functional group, along with a side chain (R group) that varies between different amino acids. They are the monomers of proteins, as mentioned in the script, and are essential for understanding the basic structure of proteins.

💡Peptide Bond

A peptide bond is a covalent chemical bond that links two peptide molecules, formed between the carboxyl group of one amino acid and the amine group of another. The script refers to peptide bonds as the linkage that connects amino acids in a polypeptide chain, which is a fundamental aspect of protein structure.

💡Spectrophotometry

Spectrophotometry is an analytical technique that measures the amount of light absorbed or transmitted by a substance at a specific wavelength. In the video, visible light spectrophotometry is used to measure the protein-Bicinchoninic acid (BCA) complex, which is indicative of protein concentration.

💡Glutathione S-Transferase (GST)

GST is a group of enzymes that play a crucial role in detoxification processes by catalyzing the conjugation of reduced glutathione to various hydrophobic compounds, making them more soluble and easier to excrete. The script discusses the importance of measuring GST activity as part of the experiment.

💡1-Chloro-2,4-dinitrobenzene (CDNB)

CDNB is a chemical compound used as a substrate in assays to measure GST activity. The script describes the use of CDNB in the experiment to assess the enzyme's ability to catalyze the formation of a conjugate with glutathione, indicating GST activity.

💡Bovine Serum Albumin (BSA)

BSA is a protein commonly used as a standard in protein assays due to its well-characterized properties. In the script, BSA is used to create a standard curve in the Bradford method, allowing for the determination of protein concentrations in the experimental samples.

💡Xenobiotics

Xenobiotics are foreign substances, such as drugs or environmental chemicals, that are not naturally produced by an organism and can be harmful. The script discusses how GST enzymes are involved in the detoxification and excretion of xenobiotics from the body.

Highlights

Experiment aims to determine the specific activity of Glutathione S-Transferase (GST) enzyme and protein concentration using the Bradford method.

GST enzyme activity in protein mixtures depends on the amount of protein used, necessitating protein content measurement.

Proteins are built from amino acids, which are the monomers, and understanding their structure is crucial for the experiment.

Different methods for protein quantitative analysis are discussed, including volumetric, gasometric, and spectrophotometry.

The Bradford method, a type of spectrophotometry, is chosen for its simplicity and speed in measuring protein concentration.

The Bio-Rad protein assay is highlighted for its advantages in measuring protein-peptide chain complexes.

GST enzyme plays a critical role in phase 2 detoxification processes in the body.

The experiment involves five steps: preparation of cytosolic fraction, enzyme activity determination, standard curve preparation, protein content determination, and specific activity calculation.

Cytosolic fraction containing GST is prepared from mice liver using a series of centrifugation steps.

GST enzyme activity is measured using the CDNB method, which involves a reaction with GSH and monitoring absorbance.

A BSA standard curve is essential for determining the concentration of the GST enzyme in the sample.

The Bradford method is used to measure the protein concentration in the cytosolic fraction containing GST.

Calculating enzyme specific activity is crucial for understanding the enzyme's purity and efficiency.

The significance of measuring enzyme activities in pharmaceutical fields, especially in cancer treatment, is discussed.

GST enzyme's role in detoxifying chemotherapeutic agents and its potential as a target for cancer treatment is highlighted.

Transcripts

hello and welcome to the second

experiment called the determination of

enzymes specific activity

the objectives of these experiments are

determine the gst enzyme activity and

determine the protein concentration by

the right method and the last one is

determine determine the specific

activity of dsd enzyme

the fundamental of these experiments are

the activity of the gst enzyme in the

protein mixtures

depends on the amounts of protein used

in the reaction therefore it is

necessary to know that the protein

content in the edit folio

so as you can see here on the picture

there is a monomer and polymer here

so the monomer is a small molecule let's

say this monomer contains a two white

circle and

one black circle

while the polymer is

is a long chain molecule made up of

repeated patterns of monomers for

example here there are four

uh different uh four repeated patterns

of monomer containing white white and a

black circle

so a monomer is a single molecule that

can be joined together with other same

molecules to form a polymer

the building blocks of the proteins are

amino acid which contain elements such

as

hnoc

and more

they are the monomers of the proteins

when hundreds or thousands of amino

acids join together they create the

proteins which are then used for many

tasks in organisms such as doing work in

cells help with that with dna

replication and so on

so we can

say that monomer is an amino acid itself

and the polymer is a protein

so amino acid there are 20 kinds of

amino acid that differentiate by the r

chain here

there is an amine chain and the carboxyl

chain here and the r can be anything so

it can be a non-polar it can be

uncharted polar or a chart polar

so amino acid itself can polymerize to

form a linear chains for example like d

peptide here at the bottom one

so these two

amino acids are

joined together with the peptide bond

here

so it can be said that this is a d

peptide

so moving on so uh

how to quantify the protein so and

protein quantitative analysis there are

three types of it so there is a

volumetric and the gastometry and sp

spectrum for dometry and the volumetric

analysis containing there is a dou haul

which is uh the determination of the

total end and then formal titration

there so they are using the

uh alkali metric titration and a

gasoline while the gasometric analysis

they used uh converting the an

amine group to an n gas

while the spectrophotometry here so

there are two types of spectrophotometry

the uv and the visible light the uv is

used used for

uh quantified proteins with aromatic

groups such as phenylalanine thyrosine

and riptophan while the visible light

such as your buret falling and lorry

in

the

way

so in our

in our experiment here we are going to

use the visible light spectrophotometry

we are using the bio-red uh

analysis so the bare analysis is measure

[Music]

the protein complex between the peptide

chains uh that

making a complex with the

buret

ion such as

co2 plus here

and their protein complex uh between the

protein and by red reaction here co2

plus will be performed in a blue purple

color which is can be measured under the

wavelength 550 nanometer

so the biored reaction gives a

plus reaction to a compound containing

ch2 and h2 or chn hn h2 rcs and h2

the advantages of using the biored

method is that the concentration of a

protein is a comparable to the

polypeptide bone

and then the method is uh

simple and fast all the disadvantages of

the bird method here is they are less

sensitive

and then the color is sometimes an

unstable yeah

so why this method is less sensitive to

others because

many a sample that containing like this

uh

here chains ch2 and several this for

this kind of chain so they can be

measured by this method

so

it can be protein it can be others uh

sample so that's why it's less sensitive

so moving on why

a big questions

why we need to measure the protein

concentration and why we need to measure

the enzyme activities is that really

important

so for our experiment we are going to

[Music]

measure the gst enzyme so as you can see

here in the picture the phase two enzyme

reaction

there uh involves three different

enzymes such as

glucoronaceal transferase or ugt

in glucoronidation

while the second one is

enzyme sulfur transferases or st

involving in sulfation and the last one

is enzyme glutathione as transversal

involved in glutathione conjugation

so we're going to

explain about the

dst here or glutathione s transparency

this phase 2 enzyme reaction are

involved

in the excretion of some xenobiotics in

our body

so including the gst one

so as we can see here in the picture the

glutathione and enzyme biotic

helped by the enzyme of tst will

transform or make a glutathione s

conjugate to be ex

to be excluded

from our body

for the glutathione conjugation as i

already explained to you that there are

three enzymes there so the glutathione

conjugation

uh include and

several uh

xenobiotics such as electrophilics

antibiotics or the synovitics that

already uh transform bio biologically to

electrophilic trophies

while the substrate for the gst itself

share three common features

such as the

the substrate must be hydrophobic and

the electrophilic and react to

non-enzymatically with potassium

at a measurable rate

and so moving on to the next slide there

are

three types of uh

gst

conjugation or

there is three different processes

for making the godathion s conjugate or

conjugation the first one is a direct

conjugation by displacement of an

electron uh withdrawal

uh withdrawing group so

sorry it's covered by this screen here

so for example there is a four nitro

quinoline one oxide here on the left

so this

nitro oxide

group

will be

displaced by the gs minus one so that

this position will be displaced by the

gs while the direct conjugation by

adding of glutathione here means that

for example the

j ethyl malaya here

the the h plus ion or the double bond

here in the c

will be adding uh or will be

broken down uh

by the gs so the c uh double bond will

be

replaced by the c with the gs ion here

while the conjugation of a strain ring

system uh format metabolically for

example this is a chlorobenzene or

p450 will be

a

form

or uh

will be transformed into

a 3 4 oxide which is a structurally

this is

more

uh

unstable

uh by p 450 so that the gs minus or

glutathione minus here can be

displaced or can be

replace

the h

ion here by the

gs ion

as you can see here there are several

types of gst

detoxification or types there is a alpha

and fee in mu and tether and fee and

zeta and so on

and the substrate and the gst enzyme

firstly published in 1974

including the cdnb that you are going to

use in your experiment

so uh moving on uh the preparation upon

determination of enzyme specific

activity measurement so in this

experiment you are going to do uh five

different things or five uh continue

i mean related uh experiment the first

one is prepare the cyto cytosolic

fraction containing the gst

enzyme from mice this is already

prepared by

the

uh by our staff and the second one is

the determination of enzyme activity or

gst in a red liver by the cdnb method

and the third one is a preparation of

provine serum albumin standard curve

using the buyer method and the fourth

one is determination of the protein

content of cytosolic fraction containing

gst by the bird method and the last one

you're going to determine the enzyme

specific activity calculation so overall

there are five steps to

finally can

measure the specific activity of the dst

enzyme

so for the first preparation cytosolic

fraction of the gst from mice

if we can

read here

we have

we we need to fit the male rats which is

the strain sprocket only one with the

weight 200 and up to 250 with the pellet

and water until it full and let the red

fast for

24 hours prior to the next step then

after that you need to sacrifice the red

with the neck and leave with the neck

dislocation

and for the cytosolic fraction

preparation containing dst by a

centrifuge method so after you

did the dislocation so

the liver will be uh

chop up and then will be uh

will be uh separate between the enzyme

and the the other fraction by using the

centrifuge centrifugation method

and then after that the red liver take

the red lever and immediately

uh put and phosphate

buffer uh in the cold one and add ph

seven then measure the weight and cross

the red liver in a phosphate buffer

using a coat blender

for five minutes

and after that centrifuge the refined

homogeneity at 10 000 g for 30 minutes

at 44 degrees

and take an end transfer the supernatant

to a new tube and discharge the

precipitated layer

and centrifuge the supernatant at

105

g 1000g for 90 minutes at 4 degree

it's like a

several step of centrifugation to take

the tst enzyme

and the supernatant layer is cytosolic

fraction containing the gst enzyme as i

already explained before

after the centrifugation there's

supposed to be two layers there so here

we need to take and out the supernatant

and store the cytosolic fraction at

minus 80 until for the use for the cdnb

conjugation with gsh

and the first step that already prepared

by our laboratories so the second step

is we're going to determine the enzyme

activity or gst right liver by the cdnb

method cdnp is stand for one chloro 2.4

d nitrobenzene

so here as we can see on the table there

are

two different fraction or two different

type one is a

dst

cytosol fraction one is a an arc or it's

a distilled water

so

why we need to use the

distilled water because it's going to be

used as a blank here

so please prepare two types of of lust

there

[Music]

as you can see on the explanation of the

experiment

you are going to prepare two flags one

is cytosol fraction

and then the other one is distilled

water and put 920 for split buffer and

then

put 20 microliter of gsh and 20

microliter of cdnb

and then the last one is add the

cytocytosol fraction for example and

distill water for blank the gsdnb

conjugate when

when after you mix it up all of this

reagent you you will have a gsd and b

conjugate product form and then measure

at 340 nanometer from minute 0 to minute

3 using a spectrum photometric as a

simple kinetic program so it means after

you

mix all the reagent there so you need to

uh measure

the zero minute and

minute three so you have two different

uh

measurement

results for each sample so you will you

will have

four different measurement there

uh the one two for example and two for

plain

so please make a note that the

measurement time which is considered as

the 0 minute the same as the other

treatment it means that if you are mixed

all the reaction

so the zero minute it means the the last

time when you edit the cdnb for the

sample also similar for the blank one

so starting time is calculated yeah

after you add

at the city and b so the city and b1

should be added

the last after you put all the reaction

there

such as first cytosol fraction phosphate

buffer gsh and the last one you need to

put the cdnb and

make it as zero minute

so the end of product will be rate or

observation absorbent absorption per

minute so

after that the gst enzyme activity is

then measured in micro mole product per

minute so the observation you need to

measure there is reaction speed so it

means the absorbance the delta

absorbance so

before uh so as you can see here there

is a two uh measurement

the zero minute and the minute three so

the delta minute it means that the

minute three uh minus minute zero so you

can got the delta absorbance there per

minute

and then

the other calculation here is you need

to

calculate the gst activity and speed

reaction in b here divided by eight

point five hundred uh

[Music]

micro mole percent per micro mole

centimeter so you can got the micro mole

gstnb per minute is

stated as c

the result of this calculation is needed

for

further calculation the activity of the

enzyme so you need to

measure all of this step

and the third

one after you

measure the

speed of

speed of reaction so the third one is

making a standard curve why you need to

make a standard curve because you need

to

measure the concentration of the gst

enzyme so

here is how to measure

how to make the bsa standard curve or a

bsa stand for the bovine serum albumin

using the biored method

so the first materials is biored reagent

and the standard solution of bsa and the

phosphate buffer you can read by

yourself and then the procedure is very

simple so wait

the albumin and put it in a

volumetric flask and add little quartz

and make an albumin standard curve with

the series concentration ranging from

100 to 700 microgram per milliliter so

the procedure in c here is very

important because you need to make a

range of concentration that you're going

to use to make a standard curve

so here it is the standard curve table

here

so there is one up to six and the other

one as

a sample so it's going to be a seven

different uh flask or seven different

solution you need to prepare

the first one is a albumin solution

which is contained the bsa one microgram

per milliliter and the second plus you

is the vsa concentration of 1.5 and the

other one is 2 and 3 and 4 and for the

last one

there is no bsa there because it's going

to be your blank

and then the last one is your sample

of gst sample that you are going to

figure out how many uh how many

concentration is there

so

the standard curve

will be

making this uh

six

uh

six absorbance uh reading

and you need after you read the

absorbance there so you need to make a

linear graph equation

uh like this is the concentration this

is the absorbance there so you you're

going to find out

the the concentration of the sample you

are added

uh you are going to measure so for

example

uh

or after you mix all of the reaction to

make the standard curve and then you got

the equation like for example

y

uh

y equal to b x plus a

and for example if

the

absorbance is y so if the absorbance is

0.234

then the

0.234 equals bx plus a

and the x1 stands for your concentration

your sample concentration so it means

that

0.234 which is y minus a divided by b

so the four

step is the determination of protein

concentration of cytosol fraction

containing the gst

so here

also uh all you need to do is just mix

mix all of the reagent here cytosol

fraction phosphate buffer and

bioreaction

and incubate those solution in the room

temperature for 10 minutes and measure

the absorbance absorption of the mixed

solution at 550 nanometer

against a blank

which is containing no cytosol fraction

and then

after you read all of the this

absorbance here this this this one this

absorbance here

you're going to

measure the cytosol protein content

which is the total uh

the total volume per cytosol fraction

milligram per microliter

and this one is you can get this one by

uh measuring this one

first

and within within the form microliter

there were going to be

some milligram protein so it means after

you

you

measure the eggs

uh protein concentration which is shown

here you can put the concentration as a

cytosol protein contained by dividing

the

ten thousand uh the thousand

one

times the total volume uh divided

cytosol fraction volume

and then within the four microliter

there they are going to be

d the cytoso protein contain within a

four microliter they're going to be a

milligram protein

so please make a note the one unit of

enzyme is the total of enzyme that

catalyze one microliter of substrate

reaction per minute and an enzyme

specific activity is the total of enzyme

units per milli divided by the protein

concentration in milligram per milli or

the total enzyme unit over milligram

protein therefore the unit for specific

enzyme activity is unit per milligram

and then the purity of enzyme is

proportional to the value of its

specific activity meaning that the

greater value of the specific activity

the higher the purity of the enzyme

so the next question is why we need to

measure the enzyme activities regarding

to the pharmaceutical fields why

um ensuring the gst enzyme is so

important

for example here

there is a

one

scientific report

here

with the title advance in antitumor

agents start getting glutathione as

transversal

so the dst is as already explained to

you that yes it's a family office 2

detoxification enzyme

so

[Music]

the this enzyme

catalyzed the conjugation of glutathione

including the chemotherapetic agents

so

the

tst is

an enzyme that will be excluded

or catalyze

or will be ex will be will will help the

excretion of some semibiotics

so

the gst in a

[Music]

cancer patient that as you can as you

can as you know that the cancer is

such a cell which is uh

have a super

growth that they can uh divide the cell

can divide into a multiple or trill

multiple

uh produce multiple cell so

you can imagine that the tst enzyme is

often produced there so it means

the excretion of some xenobiotics will

be totally also

uh

multiple times ex

doing excretion

so it means that

whenever the cancer patients

they receive some chemotherapy

so it means

the cell will be

expressed or will be exclude

the chemotherapetics drugs also in huge

amounts

so the gst enzyme

can be

can be targeted as an

uh

xenobiotic target

so

whenever the gst is inhibited so it

means that the chemotheraphatic

drug can be used or can be treated the

cancer patient successfully because the

gst is inhibited and the gst cannot

express or exclude

any

chemotherapeutic drugs outside our body

so it means that the chemotheraphatic

drugs

can

can give any benefit a maximum benefit

to our body

so thank you for listening