The development of a Department of Biochemistry at the University of Louisville began in 1918 with the recruitment of Alfred W. Homberger as the Chairman of Physiological Chemistry. For many years, he served as Chair of both the Departments of Chemistry and Physiological Chemistry. In 1942, he moved his office to the Medical school to devote his full-time efforts to the dental and medical teaching program. Professor Homberger and his small staff maintained a strong teaching program in the School of Medicine until he retired in 1951. In 1952, the University of Louisville recruited a vigorous, young scientist, Dr. John F. Taylor, from Nobel Prize laureate Dr. Carl Cori's department at Washington University, St. Louis, to chair the department. Dr. Taylor received his Ph.D. from Johns Hopkins University and pursued postgraduate training at Harvard with Professor A. Baird Hastings. During his career at the University of Louisville, Dr. Taylor published extensively in the area of hemoglobin structure and function. In addition, he is credited with the first successful crystallization of the enzyme, aldolase.

In 1972, Dr. Taylor stepped down as Chairman of the Department and Dr. Robert McGeachin was appointed as Acting Chairman. Dr. McGeachin was a master educator, teaching carbohydrate metabolism to medical and graduate students. He introduced students to the biochemical basis of diabetes mellitus, and received the Golden Apple Award, awarded annually by the medical students to the master educator of the year. He was fondly known as Dr. Sweets by the medical students. In 1976, Dr. James L. Wittliff from the Department of Biochemistry at the University of Rochester was named Chairman of Biochemistry and brought a large research group focused on biochemical endocrinology to the medical campus. Dr. Wittliff's research program involves studies on the characterization of the receptors for steroid hormones and their regulation during oncogenesis. A number of new faculty were recruited and several of these faculty continue to strengthen the Department.

In 1986, Dr. Russell A. Prough, from the Department of Biochemistry, Unversity of Texas Southwestern Medical School at Dallas, accepted the chairmanship of the Department of Biochemistry. His research program centers on the mechanism of action and regulation of cytochrome P-450 and other drug metabolizing enzymes. In Academic Year 1987-88, Nancy C. Martin, Ph.D., was recruited from the Department of Biochemistry, the University of Texas Southwestern Medical Center, Dallas, Texas to assume the Preston Pope Joyes Chair of Biochemical Research. Her research interests are in the area of yeast molecular biology and transport of nuclearly encoded proteins into the mitochondria of that organism. Six other faculty members were recruited to the Department between 1987 and 2003. All of these individuals have extramurally supported research programs and have provided the Department with additional research strength in molecular biology and in the graduate and professional teaching programs of the Department.

In February 2003, Dr. Kenneth S. Ramos was named Chairman. Dr. Ramos came to the Department from Texas A & M University where he had served as Professor in the Department of Veterinary Physiology and Pharmacology since 1995. His research interests center on gene-environment interactions in human health (chemical atherogenesis, glumerulo-nephropathies, and nethrogenesis) and redox-regulated gene expression (GSTA1, osteopontin, WT-1, L1Md, grp 78, and c-Ha-ras). Five tenure track and two term track faculty were recruited to the Department since Dr.Ramos' arrival.

In September 2007, Dr. Ramos stepped down as Chair to pursue additional opportunities in the School of Medicine. Dr. Prough was appointed as Acting Chair until April 2009.

On May 1, 2009, Dr. Ronald Gregg, PhD was appointed as Chairman of the Department of Biochemistry and Molecular Biology. Dr. Gregg received his PhD from the Biochemistry Department at the University of Queensland, Australia. He pursued post-doctoral studies with Dr. Oliver Smithies at the University of Wisconsin, Madison. In 1997 he joined the faculty of the Department of Biochemistry and Molecular Biology at the University of Louisville, rising through the ranks to full professor in 2007.