Project Info
Structure and Function of Nitrile Hydratases
Richard Holz
rholz@mines.edu
Project Goals and Description:
The goal of this project is to explore the reaction pathway of Nitrile Hydratase (NHase). NHase is used as a catalyst to convert nitriles into amides. Amides have extreme significance from a biological standpoint and appear in peptides, DNA, and RNA. Amides also participate in processes that are antibacterial, antifungal, and anthelmintic. This powerful metalloenzyme, is found in several microorganisms and plants and plays a key role in several biological systems. However, the reaction pathway for the catalysis of nitriles to amides via NHase is still being explored. The study of NHase may bring important advances for “Green Chemistry” and for industry. NHase could be exploited for the stereoselectivity it displays, which may have significant applications in pharmaceuticals and has great potential as a catalyst for organic chemical processing. NHase can also be used as a greener method to synthesize polyacrylamide, which is a precursor for plastics that are widely used. This project will utilize methods such as spectroscopy and crystallography to aid in understanding the catalytic mechanism of NHase. Specific approaches, which look at the role of the metallochaperone, the structural difference in subunits, and the electronic differences in the metal center of the protein will be coupled with data to provide insight into the biological role of NHase.
More Information:
Grand Challenge: Provide access to clean water.
https://chemistry.mines.edu/project/holz-richard-c/
Primary Contacts:
Richard Holz, Rholz@mines.edu
Callie Miller, cgmiller@mines.edu
Student Preparation
Qualifications
At least JR status and either a Chem, QBE, or CBE major.
TIME COMMITMENT (HRS/WK)
10
SKILLS/TECHNIQUES GAINED
The goals for this student will be to develop a strong foundation in biochemistry and relate it to larger industrial and the environment applications. The student will gain an understanding of important biological processes by working with BL21 cell cultures to produce protein as well as molecular biological techniques to produce mutated enzymes. These enzymes will then be purified using a Nickel NTA column, with which the student will become very familiar along with enzyme kinetics. Through this experience, the student will build progressive knowledge of how to read and interpret chromatography and kinetic data. By the end of the MURF project, the student will be able to present their research in an intelligent and relevant manner by focusing on the importance of nitriles and amides as they relate to biochemistry.
MENTORING PLAN
The student will work with a senior graduate student and met with me once a week to discuss their results and future experiments. The will be mentored on presentations of their work including oral, poster and journal article preparation.
PREFERRED STUDENT STATUS
Junior
Senior