Production of human parathyroid hormone from microorganisms

Abstract

The invention provides recombinant plasmids containing in DNA sequences coding for human preproparathyroid hormone. The invention further provides microorganisms, for example E. coli, transformed by these plasmids. The invention also provides a plasmid for insertion into yeast and a transformed yeast in which the plasmid contains DNA coding for parathyroid hormone. Parathyroid hormone is then secreted by the transformed yeast. Further the invention provides alternate polypeptides having parathyroid hormone activity, including PTH analogs, fragments and extensions, and provides alternate leader sequences and secretion signal sequences which can be used in the present invention. Finally, there are provided methods for purification of the secreted PTH hormone and/or derivatives.

We claim:  
     
         1 . Essentially pure recombinant hPTH.  
     
     
         2 . Substantially pure synthetic hPTH.  
     
     
         3 . Substantially pure recombinant hPTH.  
     
     
         4 . The substantially pure recombinant hPTH of  claim 3  wherein said hPTH is at least about 90% pure.  
     
     
         5 . The substantially pure recombinant hPTH of  claim 4  wherein said hPTH is at least about 95% pure.  
     
     
         6 . Substantially pure recombinant hPTH which is resistant to degradation by a KEX2 like proteolytic enzyme.  
     
     
         7 . A substantially pure hPTH derivative which is both intact and exhibits native biological activity.  
     
     
         8 . A substantially pure hPTH derivative which is resistant to degradation by a KEX2 like proteolytic enzyme and which is both intact and exhibits native biological activity.  
     
     
         9 . A genetically engineered microorganism capable of expressing an intact hPTH.  
     
     
         10 . The microorganism of  claim 9  wherein said organism is yeast.  
     
     
         11 . A substantially pure intact hPTH, obtained by expression and secretion of said hPTH from a genetically engineered-microorganism.  
     
     
         12 . The substantially pure intact hPTH of  claim 11 , wherein said hPTH is resistant to degradation by a KEX2 like proteolytic enzyme.  
     
     
         13 . The substantially pure intact hPTH of  claim 11 , which is obtained by a purification step after expression and secretion.  
     
     
         14 . The substantially pure intact hPTH of  claim 12 , which is obtained by a purification step after expression and secretion.  
     
     
         15 . The substantially pure intact hPTH of  claim 11 , wherein said genetically engineered microorganism is yeast.  
     
     
         16 . A substantially pure intact hPTH derivative, obtained by expression and secretion of said hPTH from a genetically engineered microorganism.  
     
     
         17 . The substantially pure intact hPTH derivative of  claim 16 , wherein said hPTH is resistant to degradation by a KEX2 like proteolytic enzyme.  
     
     
         18 . The substantially pure intact hPTH derivative of  claim 16 , which is obtained by a purification step after expression and secretion.  
     
     
         19 . The substantially pure intact hPTH derivative of  claim 17 , which is obtained by a purification step after expression and secretion.  
     
     
         20 . The substantially pure intact hPTH of  claim 16 , wherein said genetically engineered microorganism is yeast.