I do not think so, if you freezed the bacteria in glycerol.
After storing the bacteria you will have to cultivate them again on the agar. The bacteria will live their life and excrete the same substances as usually. I recommend to not use the first culture after freezing for the tests. The best option will be third recultivation. So there should not be any influence of the glycerol on them. I worked on bacteria and freezed them in glycerol. After some time I used this same bacteria as I described above for the tests (catalase, oxidase, API tests) and it works. Good luck with your research :)