Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1997-07-03
2000-01-04
Railey, II, Johnny F.
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
536 237, C12N 1531, C07K 14195
Patent
active
060111445
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
The present invention relates to a process for the production of poly(hydroxy acids) by means of recombinant bacteria which contain and express at least one fragment of the gene of poly(hydroxy fatty acid) synthase from Thiocapsa pfennigii and which are selected from the group comprising: Pseudomonas putida GPp104 (pHP1014::E156), Alcaligenes eutrophus PHB 4 (pHP1014::EIS6), Pseudomonas putida GPp104 (pHP1014::B28+) [DSM #9417] and Alcaligenes eutrophus PHB 4 (pHP1014: B28+) [DSM #9418], whereby the bacteria are cultivated in a mineral medium under aerobic conditions, whereby one offers the bacteria at least one substrate carbon source which is selected from the group consisting of: levulinic acid, salts of levulinic acid, esters of levulinic acid, lactones of levulinic acid, substituted levulinic acid or, as the case may be, its derivatives; 5-hydroxyhexanoic acid, its salts, esters and lactones; 4-hydroxyheptanoic acid, its salts, esters and lactones; 4-hydroxyoctanoic acid, its salts, esters and lactones, their halogenated derivatives as well as their mixtures; one incubates the bacteria for a certain time with the carbon source; and one isolates the poly(hydroxy fatty acid) polymers that have been synthesized by the bacteria; bacterial strain is selected from the group which comprises Pseudomonas putida GPp104 (pHP1014::B28+) [DSM #9417] and Alcaligenes eutrophus PHB 4 (pHP1014::B28+) [DSM #9418]; processes; of the poly(hydroxy fatty acid) synthase from Thiocapsa pfennigii characterized by the feature that it has at least the nucleotide sequence of sequence sections 180 through 1280 (phaE) and 1322 through 2392 (phaC) of the DNA sequence SEQ ID NO: 1.
In this age of increasing environmental awareness, there are increasing attempts in industry and science to produce biodegradable polymers. In this regard, these new types of environmentally compatible polymers should essentially have the same properties as those polymers which, for decades, have been prepared via organic chemical synthesis.
In particular in this connection, the ability to process the new types of biodegradable polymers ought to be provided in a similar manner to the processing of conventional plastics using the same methods such as, for example, extrusion, injection molding, injection compression, foaming, etc.
A big disadvantage of organically synthesized plastics is, however, that many of these plastics have enormous biological half-lives or, as the case may be, they cannot be disposed of in garbage dumps or in garbage incineration plants in a non-harmful manner but, rather, aggressive gases are frequently produced such as, for example, in the case of poly(vinyl chloride) which liberates hydrogen chloride gas during incineration.
A first step in the direction of success with environmentally compatible materials was achieved by means of synthetic substances, e.g. the paraffin-like polymers polyethylene and polypropylene since these essentially release only CO.sub.2 and water on incineration.
In addition, many attempts have also been made by means of so-called replaceable raw materials such as, e.g. plants that contain a lot of polysaccharide such as potatoes, corn, wheat, beans, peas or similar materials, to obtain the naturally occurring polysaccharides in these plants and to prepare polymers from them which are usable in plastics technology and which are biodegradable.
However, in the case of such polymer materials comprising replaceable raw materials, one is essentially relying on the natural quality of the polymers that occur in these higher plants and only the relatively complex processes of classical cultivation and modern gene technology offer themselves for modification at the genetic level.
An essential further step in the direction of naturally occurring polymers, which are very similar to synthetic thermoplastics, was brought about by the discovery of poly(3-hydroxybutyric acid) by Lemoigne in 1926 [Lemoigne, M. (1926) Products of the dehydration and polymerization of .beta.-oxybutyric acid, Bull
REFERENCES:
Liebergesell et al., Appl. Microbiol. Biotechnol. 40:292-300 (1993).
Valentin et al., Appl. Microbiol. Biotechnol. 40:710-716 (1994).
Kimmel., "Identification and Characterization of Specific Clones: Strategy for Confirming the Validity of Presumptive Clones," in Methods in Enzymology: Guide to Molecular Cloning Techniques, vol. 152, Academic Press: New York, pp. 507-511 (1987).
Greene et al., "Subcloning," in Methods in Enzymology: Guide to Molecular Cloning Techniques, vol. 152, Academic Press: New York, pp. 512-522 (1987).
Barnes., "Sequencing DNA with Dideoxyribonucleotides as Chain Terminators: Hints and Strategies for Big Projects," in Methods in Enzymology: Guide to Molecular Cloning Techniques, vol. 152, Academic Press: New York, pp. 538-556 (1987).
New England Biolabs Catalog 1992. New England Biolabs, Beverly, MA. p. 32.
Liebergesell Mathias
Pries Andreas
Steinbuchel Alexander
Valentin Henry
Bond Gary
Monsanto Company
Railey II Johnny F.
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