This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Brazzein is a small, highly heat stable sweet protein. We have overexpressed and purified brazzein in E. coli. Our main goal is to determine the sweet sites on this small protein. In this study, we are using five different brazzein variants with either increased or decreased sweet property and would like to see if there are local or global changes due to these mutations. Furthermore, we would like to examine these mutations in more structural details and relate those (if there is any significant change) to their sweetness property for the future structure/function studies in addition to making more potent brazzein
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