Carbapenemases for use with antibiotics for the protection of the intestinal microbiome

Abstract

This invention relates, in part, to various compositions and methods for protecting the gastrointestinal microbiome from antibiotic disruption.

1 - 76 . (canceled) 
     
     
         77 . A method for producing a broad spectrum recombinant carbapenemase in  Escherichia coli  ( E. coli ), comprising:
 (a) providing a host  E. coli  cell transformed with a vector comprising a nucleic acid sequence encoding the carbapenemase operably linked to an inducible promoter;   (b) culturing the  E. coli  cell in a cell culture medium comprising zinc and inducing the expression of the carbapenemase; and   (c) recovering the carbapenemase from a soluble fraction prepared from the cytoplasm or periplasmic space of the  E. coli  cell,   wherein the culturing is in the presence of about 100 μM zinc, such that the amount of carbapenemase in the soluble fraction is increased and the amount of carbapenemase in inclusion bodies is reduced, relative to culturing in the absence of zinc.   
     
     
         78 . The method of  claim 77 , wherein the method yields at least 10 grams of substantially active carbapenemase per liter of culture. 
     
     
         79 . The method of  claim 77 , wherein the method yields at least 15 grams of substantially active carbapenemase per liter of culture. 
     
     
         80 . The method of  claim 77 , wherein the recovered carbapenemase is soluble. 
     
     
         81 . The method of  claim 77 , wherein the culturing is in a bioreactor or a shake flask. 
     
     
         82 . The method of  claim 77 , wherein the  E. coli  cell is induced to express the carbapenemase using isopropyl β-D-1-thiogalactopyranoside (IPTG).