Production of human parathyroid hormone from microorganisms

A process for producing recombinant parathyroid hormone (PTH) (1-84) protein comprising the steps of:

• (a) providing a yeast microorganism comprising:
• (1) a first DNA sequence encoding the first nineteen amino acids of the Saccharomyces mating factor .alpha.1 as a leader sequence;
• and
• (2) a second DNA sequence encoding PTH (1-84) protein,
• wherein the DNA sequence encoding the leader sequence and the DNA sequence encoding PTH (1-84) protein are operably linked;
• (b) culturing said yeast microorganism to allow expression of said PTH (1-84) protein, thereby producing PTH (1-84) protein, wherein the yeast microorganism cleaves the PTH (1-84) protein from the leader sequence upon secretion of the PTH (1-84) protein from the yeast microorganism;
• and
• (c) purifying the resultant PTH (1-84) protein.

A process for producing recombinant parathyroid hormone (PTH) (1-84) protein comprising the steps of:

• (a) providing a yeast microorganism comprising:
• (1) a DNA encoding a leader sequence;
• and
• (2) a second DNA sequence encoding a peptide that is a derivative of PTH (1-84) protein, wherein the cleavage site after the pair of basic amino acids at positions 25 and 26 of the derivative PTH (1-84) protein has been modified such that the hormone is excluded as a substrate for yscF protease, wherein the DNA encoding the leader sequence and the DNA sequence encoding PTH (1-84) protein are operably linked;
• (b) culturing said yeast microorganism to allow expression of said PTH (1-84) protein derivative, thereby producing PTH (1-84) protein, wherein the yeast microorganism cleaves the PTH (1-84) protein from the leader sequence upon secretion of the PTH (1-84) protein from the yeast microorganism;
• and
• (c) purifying the resultant PTH (1-84) protein.

A process for producing recombinant parathyroid hormone (PTH) (1-84) comprising the steps of:

• (a) providing a yeast microorganism comprising:
• (1) a DNA sequence encoding PTH (1-84) protein;
• and
• (2) a second DNA sequence encoding a signal sequence, wherein the signal sequence has the following:
• (i) a positively charged amino-terminal;
• (ii) a hydrophobic core region;
• and
• (iii) a polar COOH-terminal region,
• wherein the signal DNA sequence and the DNA sequence encoding PTH (1-84) protein are operably linked;
• (b) culturing said yeast microorganism to allow expression of said PTH (1-84) protein, thereby producing PTH (1-84) protein, wherein the yeast microorganism cleaves the PTH (1-84) protein from the signal sequence upon secretion of the PTH (1-84) protein from the yeast microorganism;
• and
• (c) purifying the resultant PTH (1-84) protein.

A process for producing recombinant parathyroid hormone (PTH) (1-84) protein comprising the steps of:

• (a) providing a yeast microorganism comprising:
• (1) a DNA sequence encoding PTH (1-84) protein;
• and
• (2) a second DNA sequence encoding a functional signal sequence having the following sequence
• Met-Asn-lie-Phe-Tyr-Ile-Phe-Leu-Phe-Leu-Ser-Phe-Val-Gln-Gly-Thr-Arg-Gly,
• and wherein the DNA encoding the signal sequence and the DNA sequence encoding PTH (1-84) protein are operably linked;
• (b) culturing said yeast microorganism to allow expression of said PTH (1-84) protein, thereby producing PTH (1-84) protein, wherein the yeast microorganism cleaves the PTH (1-84) protein from the signal sequence upon secretion of the PTH (1-84) protein from the yeast microorganism;
• and
• (c) purifying the resultant PTH (1-84) protein.