Creating a sub-domain in AWS EC2 Bitnami Wordpress instance

I'm trying to create a sub-domain for a website hosted on a single AWS EC2 instance. I used a bitnami single site wordpress image. I know I probably should've used a multisite image but I'm wondering if there's any way I could facilitate a sub-domain which will in no-way be associated or linked to or conflicting with the current Wordpress installation?

I'm using Route 53 for DNS hosting. I've tried to configure virtual hosting but it didn't work out and I think bitnami has some way of managing virtual hosting itself, I'm unsure.

Any help would be greatly appreciated.