ES2186574A1 - Nucleotide sequence that codes for a pseudomonas putida phaf protein - Google Patents

Nucleotide sequence that codes for a pseudomonas putida phaf protein

Info

Publication number
ES2186574A1
ES2186574A1 ES200102240A ES200102240A ES2186574A1 ES 2186574 A1 ES2186574 A1 ES 2186574A1 ES 200102240 A ES200102240 A ES 200102240A ES 200102240 A ES200102240 A ES 200102240A ES 2186574 A1 ES2186574 A1 ES 2186574A1
Authority
ES
Spain
Prior art keywords
protein
phaf
pseudomonas putida
fusion protein
granules
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
ES200102240A
Other languages
Spanish (es)
Other versions
ES2186574B1 (en
Inventor
Jimenez M Prieto
Tabares Cristina Moldes
Gonzalez Pedro Garcia
Lopez Jose Luis Garcia
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Consejo Superior de Investigaciones Cientificas CSIC
Original Assignee
Consejo Superior de Investigaciones Cientificas CSIC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Consejo Superior de Investigaciones Cientificas CSIC filed Critical Consejo Superior de Investigaciones Cientificas CSIC
Priority to ES200102240A priority Critical patent/ES2186574B1/en
Priority to PCT/ES2002/000454 priority patent/WO2003031624A1/en
Publication of ES2186574A1 publication Critical patent/ES2186574A1/en
Application granted granted Critical
Publication of ES2186574B1 publication Critical patent/ES2186574B1/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1241—Nucleotidyltransferases (2.7.7)
    • C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention relates to the design and production of fusion proteins with the amino-terminal end of the Pseudomonas putida PhaF protein GPo1. Said fusion proteins are produced in a strain of Pseudomonas putida which lacks protein PhaF and which has been transformed with a recombinant plasmid bearing the gene that expresses the fusion protein. In said recombinant cells, the fusion protein is immobilised for the most part in granules of medium-chain polyhydroxyalkanoate which are produced in the cellular cytoplasm when the cells are cultivated under determined conditions. Said polyhydroxyalkanoate granules which contain the fusion protein can be isolated easily by means of simple centrifugation from the cellular extract. Subsequently, the fusion protein can be easily freed from the granules without the functionality thereof being lost.
ES200102240A 2001-10-08 2001-10-08 FUSION PROTEINS IMMOBILIZED IN MEDIUM CHAIN POLYHYDROXIALCANOATE GRANULES. Expired - Fee Related ES2186574B1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
ES200102240A ES2186574B1 (en) 2001-10-08 2001-10-08 FUSION PROTEINS IMMOBILIZED IN MEDIUM CHAIN POLYHYDROXIALCANOATE GRANULES.
PCT/ES2002/000454 WO2003031624A1 (en) 2001-10-08 2002-09-27 Nucleotide sequence that codes for a pseudomonas putida phaf protein

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
ES200102240A ES2186574B1 (en) 2001-10-08 2001-10-08 FUSION PROTEINS IMMOBILIZED IN MEDIUM CHAIN POLYHYDROXIALCANOATE GRANULES.

Publications (2)

Publication Number Publication Date
ES2186574A1 true ES2186574A1 (en) 2003-05-01
ES2186574B1 ES2186574B1 (en) 2004-09-16

Family

ID=8499128

Family Applications (1)

Application Number Title Priority Date Filing Date
ES200102240A Expired - Fee Related ES2186574B1 (en) 2001-10-08 2001-10-08 FUSION PROTEINS IMMOBILIZED IN MEDIUM CHAIN POLYHYDROXIALCANOATE GRANULES.

Country Status (2)

Country Link
ES (1) ES2186574B1 (en)
WO (1) WO2003031624A1 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112391328B (en) * 2020-10-27 2022-08-05 西安交通大学 Pseudomonas putida ND6 coupled protein complex dotL gene deletion mutant strain and construction method and determination method thereof

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
GARCIA, B. et al. "Novel biodegradable aromatic plastics from a bacterial source". THE JOURNAL OF BIOLOGICAL CHEMISTRY, Vol. 274, nö 41, 8 octubre 1999, páginas 29228-29241, figura 5. *
KLINKE, S. et al. "Role of PhaD in accumulation of medium-chain-lenght poly(3-hydroxyalkanoates) in Pseudomonas oleovorans". APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Vol. 66, nö 9, septiembre 2000, páginas 3705-3710, figura 1. *
PRIETO, M.A. et al. "Engineering of stable recombinant bacteria for production of chiral medium-chain-lenght poly-3-hydroxyalkanoates". APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Vol. 65, nö 8, agosto 1999, páginas 3265-3271, figura 1. *
PRIETO, M.A. et al. "PhaF, a polyhydroxyalkanoate-granule- associated protein of Pseudomonas oleovorans GPo1 involved in the regulatory expression system for pha genes". JOURNAL OF BACTERIOLOGY, Vol. 181, nö 3, febrero 1999, páginas 858-868, todo el documento. *
VAN BEILEN, J.B. et al. "Analysis of Pseudomonas putida alkane-degradation gene clusters and flanking insertion sequences: Evolution and regulation of the alk genes". MICROBIOLOGY, Vol. 147, nö 6, junio 2001, páginas 1621-1630, todo el documento, especialmente figura 2. *

Also Published As

Publication number Publication date
WO2003031624A1 (en) 2003-04-17
ES2186574B1 (en) 2004-09-16

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