Method for the improved production and recovery of poly-.beta.-hydroxybutyrate from transformed Escherichia coli

A process for producing poly-.beta.-hydroxybutyrate comprising:

• providing a culture of Escherichia coli bacterial host cells transformed by (1) a first vector containing a DNA sequence encoding the biosynthetic pathway of poly-.beta.-hydroxybutyrate from Alcaligenes eutrophus, wherein the first vector is a plasmid selected from the group consisting of pJM9101, pJM9113, pJM9114, pJM9115, pJM9116, pJM9117, pJM9118, pJM9119, pJM9120, pJM9125, and pJM9126, and (2) a second vector containing a DNA sequence of a T7 bacteriophage lysozyme gene;
• growing the culture in a suitable medium and obtaining expression of the poly-.beta.-hydroxybutyrate biosynthetic pathway and the lysozyme gene in the Escherichia coli bacterial host cells to provide poly-.beta.-hydroxybutyrate and lysozyme;
• lysing the Escherichia coli bacterial host cells;
• isolating the poly-.beta.-hydroxybutyrate;
• and,
• collecting the poly-.beta.-hydroxybutyrate.

A process for producting poly-.beta.-hydroxybutyrate comprising:

• providing a culture of Escherichia coli bacterial host cells, in which the Escherichia coli bacterial host cells comprise a strain (S4) designated as E. coli HMS174(p4A (BstB-),pLysS), deposited with the American Type Culture Collection under ATCC Accession No. 69001;
• growing the culture in a suitable medium and obtaining expression of the poly-.beta.-hydroxybutyrate biosynthetic pathway and the lysozyme gene in the Escharichia coli bacterial host cells to provide poly-.beta.-hydroxybutyrate and lysozyme;
• lysing the Escherichia coli bacterial host cells;
• isolating the poly-.beta.-hydroxybutyrate;
• and,
• collecting the poly-.beta.-hydroxybutyrate.

A process for producing poly-.beta.-hydroxybutyrate comprising:

• providing a culture of Escherichia coli bacterial host cells transformed by (1) a first vector containing a DNA sequence encoding the biosynthetic pathway of poly-.beta.-hydroxybutyrate from Alcaligenes eutrophus, wherein the DNA sequence of the biosynthetic pathway of poly-.beta.-hydroxybutyrate is contained on a high copy number plasmid or on a runaway replication vector, and (2) a second vector containing a DNA sequence of a T7 bacteriophage lysozyme gene;
• growing the culture in a suitable medium and obtaining expression of the poly-.beta.-hydroxybutyrate biosynthetic pathway and the lysozyme gene in the Escherichia coli bacterial host cells to provide poly-.beta.-hydroxybutyrate and lysozyme;
• lysing the Escherichia coli bacterial host cells:
• isolating the poly-.beta.-hydroxybutyrate;
• and,
• collecting the poly-.beta.-hydroxybutyrate at levels of about 90 to about 95%, by weight, of dry cell weight.

A process for producing poly-.beta.-hydroxybutyrate comprising:

• providing a culture of Escherichia coli bacterial host cells transformed by (1) a first vector containing a DNA sequence encoding the biosynthetic pathway of poly-.beta.-hydroxybutyrate from Alcaligenes eutrophus, in which the DNA sequence coding for poly-.beta.-hydroxybutyrate is contained on a high copy number plasmid, and (2) a second vector containing a DNA sequence of a T7 bacteriophage lysozyme gene;
• growing the culture in a suitable medium and obtaining expression of the poly-.beta.-hydroxybutyrate biosynthetic pathway and the lysozyme gene in the Escherichia coli bacterial host cells to provide poly-.beta.-hydroxybutyrate and lysozyme;
• lysing the Escherichia coli bacterial host cells;
• isolating the poly-.beta.-hydroxybutyrate;
• and, collecting the poly-.beta.-hydroxybutyrate.

A process for producing poly-.beta.-hydroxybutyrate comprising:

• providing a culture of Escherichia coli bacterial host cells transformed by (1) a first vector containing a DNA sequence encoding the biosynthetic pathway of poly-.beta.-hydroxybutyrate from Alcaligenes eutrophus, wherein the isolated DNA sequence coding for poly-.beta.-hydroxybutyrate is contained on a runway replication vector, and (2) a second vector containing a DNA sequence of a T7 bacteriophage lysozyme gene;
• growing the culture in a suitable medium and obtaining expression of the poly-.beta.-hydroxybutyrate biosynthetic pathway and the lysozyme gene in the Escherichia coli bacterial host cells to provide poly-.beta.-hydroxybutyrate and lysozyme;
• lysing the Escherichia coli bacterial host cells;
• isolating the poly-.beta.-hydroxybutyrate;
• and
• collecting the poly-.beta.-hydroxybutyrate.

A process for producing poly-.beta.-hydroxybutyrate comprising:

• providing a culture of Escherichia coli bacterial host cells tranformed by (1) an isolated DNA sequence coding for the Alcaligenes eutrophus biosynthetic pathway of poly-.beta.-hydroxybutyrate, and (2) an isolated DNA sequence of a T7 bacteriophage lysozyme gene;
• growing the culture in a suitable medium and obtaining expression of the poly-.beta.-hydroxybutyrate biosynthetic pathway and the lysozyme gene in each Escherichia coli bacterial host cell;
• isolating the poly-.beta.-hydroxybutyrate by lysing the Escherichia coli bacterial host cells;
• and
• collecting the poly-.beta.-hydroxybutyrate.