Study of glycolysis

[dim2996]

Sorry, my english is not good. I maybe make some mistakes(

Maybe someone can answer this question:
Why is the determination and comparison of the intensity of glycolysis in different microorganisms determined in vitro, and not in vivo?
Conducting an in vivo test will not give accurate results.
The determination takes place on an intermediate: fructosodiphosphate.

Thank you

[DeoxyRiboRobert]

53 minutes ago, dim2996 said:

Sorry, my english is not good. I maybe make some mistakes(

Maybe someone can answer this question:
Why is the determination and comparison of the intensity of glycolysis in different microorganisms determined in vitro, and not in vivo?
Conducting an in vivo test will not give accurate results.
The determination takes place on an intermediate: fructosodiphosphate.

Thank you

3

I'm not entirely sure what question you're asking but I think that you may wish to consider what the objective of the study is. For example, if someone is interested in determining the quantity of glucose produced per minute or the maximum able to be produced by an organism, then it's often best to perform experiments to determine this in-vivo since there're limiting factors that are difficult to replicate in a lab environment. However, this is not applicable to every situation/organism. it may also be cheaper to perform the experiment in in-vitro which is always more enticing. 

Edited October 25, 2017 by DeoxyRiboRobert

[dim2996]

My teacher said me that we can't compare the intensity of glycolysis if we will do experiment in vivo.

We must do it only in vitro.

Сomparative analysis of the intensity of glycolysis in different microorganisms was the theme of this work.

We worked with

Pseudomonas fluorescens

Bacillus subtilis

Candida scotti

Saccharomyces cerevisiae

In the our laboratory books:

We must perform the process of glycolysis in vitro because it can be stopped at the stage of fructosodiphosphate formation.
But our teacher said that we could stop the process in vivo also.

[CharonY]

The distinction between in vivo and in vitro is a but muddied when it comes to cell cultures. What do you specifically define as in vivo? For example, if you lyse cells and measure metabolite levels, do you consider that in vivo or in vitro?

[BabcockHall]

Your use of the word intensity is not entirely clear.  Are you measuring flux, (which is basically a rate)?   

[BabcockHall]

How is glycolysis being stopped at stage of the production of fructose 1,6-bisphosphate?

[dim2996]

by adding Trichloroacetic acid

we measure optical density.

we use Seliwanoff's test

[BabcockHall]

Optical density of what, the product of Seliwanoff's test?

Do you think that you could perform the same experiment in vivo as you do in vitro?  Why or why not?

Another thing to consider: if you stop glycolysis at the stage of producing fructose 1,6-bisphosphate, what will happen to the organism?

[dim2996]

I am a student.

We made a laboratory work 'comparative analysis of the intensity of glycolysis in different microorganisms'.

our teacher asked:

why do we need to carry out the glycolysis reaction in vitro, and not in vivo?

[BabcockHall]

Please reread my previous post.  It was intended to help you answer this question.

[dim2996]

Optical density of what, the product of Seliwanoff's test?

After adding Trichloroacetic acid, we also add 0.1% Resorcinol in 95% Ethanol and 30% hcl.

 

Do you think that you could perform the same experiment in vivo as you do in vitro?  Why or why not?

Another thing to consider: if you stop glycolysis at the stage of producing fructose 1,6-bisphosphate, what will happen to the organism?

As I understood our teacher said that at the moment when we add Trichloroacetic acid to cell we will destroy them.

But we peformed experiment in such way:

1) isolation of enzymes from the cell mass;
2) carrying out the glycolysis reaction in vitro;
3) stopping the glycolysis reaction at the stage of fructose formation of 1,6-bisphosphate;
4) clarification of the filtrate;
5) determination of the concentration of fructose 1,6-bisphosphate

 

as we understood our teacher asked why we cannot

1) carrying out the glycolysis reaction in vivo;
2) stopping the glycolysis reaction at the stage of fructose formation of 1,6-bisphosphate;

3) destroy cells;
4) clarification of the filtrate;
5) determination of the concentration of fructose 1,6-bisphosphate

[BabcockHall]

You may be overthinking this.  How would you perform 2 while the cells are still intact?