I have checked this settings and it is not selected.

On the main screen of pokeys software the pin 35 is configured as output and if I open and close the mach3 the pin is changed to input and then the output not work anymore.

I think I will gave up on the pokeys because it look like it is to complicated to configure it as hobby user.

re; because it look like it is to complicated

 Yes,  they are more complicated. but they have capabilities far beyond a simple primitive step driver. every added function on a pokeys device is a HUGE advantage to a hobby CNC builder. the LCD screens, the DOZEN of outputs , the block programming language , the ic2 interface, PWM outs, these are worth spending time to learn.  the more i learn the more i respect the designers . i have 'spent' 1000s of  hours over 6 years, and I am amazed at what I can do with these.

ok back to work . .

I have now rewired the output to the pin 34 and now it is working, it looks like that the pin 35 is not useful for the output.

But during the resolving this problem I have reset setting of pokey over the polabs software and now my motors not move anymore. I'm not shure if this is the reason for problems but I heve no other idea.
The motor drivers don't get the enable signal. When I jog the axes in the mach3 the position on the screen is changing but the motors not moves.
But when I try to jog the axes I get the red light blinking on the stepper drivers. I have reinstaled everything, changed all settings which exist but nothing helped.

Is any expert on the forum who is prepared to help me for a reasonable amount of money(check my settings over the teamviewer or maybe visit me at my location near Ljubljana)?

tomy007, did you try to invert the motor enable signals in PoKeys plugin?

Also, open the IO status window (there is a button in the PoKeys plugin under IO status) - are the axes positions changing there also, but the motors don't move?

I have disconected the enable signals from the drivers and then the motors are moving.
I have trayed to inver enable signal but it does not help. Enable from the poextbus modul is all the time on 5v.

argh.... I missed the post where you wrote you are using PoExtBusOC16CNC...
In the manual for this device (https://www.poscope.com/index.php?route=product/product/download&download_id=134&product_id=81) you can see that the wire 3 is the 'motor enable signal'. Where did you connect it to?

Ah I forgot to invert the output for the enable signal, which is not connected to anything in plugin but it must be inverted.

Now the axes are moving but I thing that the movement is not smooth as it should be. When I hold the button for jog the feed rate for the axes is changing but I thing it should be constant(example: between 200-250 mm/min).
Also if I run the g-code which move the spindle in the circle I think that the movement of axes is not smooth enough.
I understand that the movement of the axis is not in real time because of the ethernet communication, but it should be better.
Can I check if anything is wrong or is that normal for ethernet communication?
Is maybe better to use pokeys only for the IOs and the standard LPT port for driving the motors?

The Pulse engine in PoKeys was designed to drive the stepper motors and it has to run as smooth as parallel port. We will be checking if there is anything wrong in the following week when the testing machine will be ready again.