Good day,
The short of it is that you'll likely need to perform a fresh reinstall. Of course, if your configuration supports it, you can vMotion VMs off the host to be upgraded so you don't suffer downtime. If your VMs are on local storage, you're looking at VM downtime.
If you want, you can back up your ESXi configuration using the vSphere CLI and vicfg-cfgbackup. It may help ease the burden of upgrading. If you have Enterprise Plus licensing, you can also use host profiles to quickly return to your original configuration.
Check out the online documentation about backing up and restoring ESXi 4.1 here
and host profiles
Cheers,
Mike Brown