The biosynthesis of the three "non-classical" clans of the beta-lactam antibiotics will be investigated. Represented by the "classical" penicillins and cephalosporins, collectively these drugs constitute 65% of the world antibiotic market and account for >$25B/yr in economic value. They remain an important mainstay of human health and longevity. But with wide spread use has come the inevitable rise of antibiotic-resistant infections. Structural modifications have slowed these effects, but increased reliance on the newer, non-classical classes, for example, the beta-lactamase inhibitor clavulanic acid and the potent, broad-spectrum carbapenems like thienamycin, will benefit from fundamental understanding of their biosynthesis for improved, lower cost manufacture by fermentation, semi-synthesis and, as we propose, chemoenzymatic synthesis using engineered biosynthetic pathways to produce clinically-used drugs and new structures with improved characteristics. The monocyclic beta-lactams are the simplest structurally but ironically the mechanism of their formation is unknown. A series of breakthroughs in the current grant period have set the stage to finally solve this problem. This understanding will guide experiments with the biosynthetically related monobactams, distinct for their N-sulfonated beta-lactam rings that confer the increasingly important property of comparative resistance to the Class B beta-lactamases (Zn++ metalloproteases). While in the current grant period we discovered and examined in detail the ATP-dependent beta-lactam forming reactions in both clavulanic acid and carbapenem biosynthesis, the essential oxidative bicyclic ring inversion processes to the antibiotic-active stereochemistry will be determined. The four known classes of beta-lactam antibiotics arise by convergent evolution and are emblematic of remarkably varied enzyme mechanisms of impressive synthetic efficiency. Research to understand them holds great training potential for students and affords the opportunity to make valuable contributions academically and practically to their manufacture and structural alteration to overcome resistance mechanisms.

Public Health Relevance

The non-classical beta-lactam antibiotics, notably the beta-lactamase inhibitor clavulanic acid and the potent, broad-spectrum carbapenems, have become increasingly important in healthcare to overcome resistant bacterial infections. Study of their biosynthesis will improve and lower the costs of manufacture, and combined chemical and enzymatic syntheses using engineered biosynthetic proteins can enable these advances and provide routes to agents with improved properties.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
High Priority, Short Term Project Award (R56)
Project #
2R56AI014937-36
Application #
8895596
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Xu, Zuoyu
Project Start
1978-09-01
Project End
2015-07-31
Budget Start
2014-08-01
Budget End
2015-07-31
Support Year
36
Fiscal Year
2014
Total Cost
$382,789
Indirect Cost
$132,789
Name
Johns Hopkins University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218