New enzymes promise cheaper, cleaner drug manufacturing

A pair of newly found enzymes might change drug manufacturing, decreasing each prices and environmental hurt.   

Researchers at Duke University School of Medicine confirmed that one of the enzymes is enough to make azetidine, an natural compound and a constructing block of a variety of therapeutics, together with antibiotics, antiviral medicine, and most cancers therapies.    

Rather than counting on harsh solvents and poisonous chemical compounds, the workforce found enzymes, PolF and PolE, that produce azetidine from a reasonable precursor compound in water.   

“The two enzymes that we discovered are functionally and structurally novel,” mentioned Kenichi Yokoyama, PhD, affiliate professor of biochemistry at Duke and senior writer of the examine. “We’re interested in applying these enzymes for biocatalytic reactions to produce structurally diverse azetidine amino acids, and other cyclic compounds.” 

A beforehand reported enzymatic course of for producing azetidine required a precursor compound that’s tough to supply and thus costly to buy; one provider sells a one-gram vial (about half the load of a U.S. penny) for $1,390. That’s about 1000-fold greater than it prices to purchase the precursor used within the Duke examine, Yokoyama mentioned.  

Azetidine is a small, “strained” ring of 4 atoms: one nitrogen atom and three carbons. It’s a problem to make due to its small measurement, Yokoyama defined. “As you make the ring smaller, you have to bend the bonds, and that requires a lot of energy. Somehow, PolF can do that job.”  

The second enzyme they found, PolE, helps by boosting the provision of a essential intermediate. 

PolF’s capabilities don’t cease there. The researchers discovered that it will possibly additionally produce a fair smaller compound, aziridine, which is a nitrogen-containing, three–membered ring.  

“One enzyme that can make both a four-membered and a three-membered ring — that’s unique and unprecedented,” Yokoyama mentioned. 

The researchers found the newfound enzymes by finding out how azetidine is made naturally in an antifungal compound referred to as polyoxin. 

Collaborators at Pennsylvania State University, together with Amie Boal, PhD, Karsten Krebs, PhD, and J. Martin Bollinger, PhD, supplied key experience in structural biology, bioinorganic chemistry, and spectroscopy, Yokoyama mentioned.  

Former postdoctoral associate Ya-Nan Du, PhD, performed a lot of the hands-on experimental work, each in Yokoyama’s lab at Duke and at Pennsylvania State, the place she discovered extra about how the PolF enzyme works through the use of stopped-flow and Mössbauer spectroscopy, which permit chemists to monitor the quick, enzyme-catalyzed reactions. 

These detailed experiments revealed the iron-containing reactive species on the middle of the PolF enzyme, liable for breaking the robust carbon-hydrogen bonds and forming the strained four- and three-membered rings, Yokoyama mentioned. 

Additional authors: Anyarat Thanapipatsiri, Jesús José Blancas Cortez, Xavier Enrique Salas Solá, and Chi-Yun Lin. 

Funding: National Institutes of Health