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ES2172380A1 - A PROCEDURE TO PRODUCE ALFA-GALACTOSIDASA RECOMBINANT THERMUS SP. T2 IN HOSPEDING CELLS. - Google Patents

A PROCEDURE TO PRODUCE ALFA-GALACTOSIDASA RECOMBINANT THERMUS SP. T2 IN HOSPEDING CELLS.

Info

Publication number
ES2172380A1
ES2172380A1 ES200000515A ES200000515A ES2172380A1 ES 2172380 A1 ES2172380 A1 ES 2172380A1 ES 200000515 A ES200000515 A ES 200000515A ES 200000515 A ES200000515 A ES 200000515A ES 2172380 A1 ES2172380 A1 ES 2172380A1
Authority
ES
Spain
Prior art keywords
alpha
galactosidase
thermus
cells
procedure
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
ES200000515A
Other languages
Spanish (es)
Other versions
ES2172380B1 (en
Inventor
Herrero Alejandro Vian
Santiago Alfonso V Carrascosa
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 ES200000515A priority Critical patent/ES2172380B1/en
Priority to PCT/ES2001/000078 priority patent/WO2001064914A1/en
Priority to AU37444/01A priority patent/AU3744401A/en
Publication of ES2172380A1 publication Critical patent/ES2172380A1/en
Application granted granted Critical
Publication of ES2172380B1 publication Critical patent/ES2172380B1/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
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2465—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on alpha-galactose-glycoside bonds, e.g. alpha-galactosidase (3.2.1.22)

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

Un procedimiento para producir alfa-galactosidasa recombinante de Thermus sp. T2 en células hospedantes. Se describe una secuencia de nucleótidos que corresponde a un gen estructural de Thermus sp. T2 ATCC 27737 que codifica para una alfa-galactosidasa (EC 3.2.1.22) termoestable. Se indica el modo de obtener una construcción genética para la expresión de dicha secuencia de nucleótidos en células de Escherichia coli mediante su clonación en un vector de expresión, con las modificaciones correspondientes en las secuencias de nucleótidos y de aminoácidos a las que la manipulación aboca. Se describe un método para obtener extractos enriquecidos con actividad alfa- galactosidasa termoestable producida en E. coli. La proteína recombinante descrita, debido a su característica de termoestabilidad, que la hace compatible con diversos procedimientos tecnológicos, tiene aplicación principalmente en el sector agroalimentario-sanitario, para la hidrólisis de alfa- galactósidos de mala digestibilidad presentes en diversos alimentos de gran consumo.A procedure for producing recombinant alpha-galactosidase from Thermus sp. T2 in host cells. A nucleotide sequence corresponding to a structural gene of Thermus sp. T2 ATCC 27737 encoding a heat stable alpha-galactosidase (EC 3.2.1.22). The way to obtain a genetic construction for the expression of said nucleotide sequence in Escherichia coli cells by means of its cloning in an expression vector is indicated, with the corresponding modifications in the nucleotide and amino acid sequences to which the manipulation leads. A method is described to obtain extracts enriched with thermostable alpha-galactosidase activity produced in E. coli. The recombinant protein described, due to its thermostability characteristic, which makes it compatible with various technological procedures, has application mainly in the agri-food-health sector, for the hydrolysis of poorly digestible alpha-galactosides present in various widely consumed foods.

ES200000515A 2000-03-03 2000-03-03 A PROCEDURE TO PRODUCE ALFA-GALACTOSIDASA RECOMBINANT THERMUS SP. T2 IN HOSPEDING CELLS. Expired - Fee Related ES2172380B1 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
ES200000515A ES2172380B1 (en) 2000-03-03 2000-03-03 A PROCEDURE TO PRODUCE ALFA-GALACTOSIDASA RECOMBINANT THERMUS SP. T2 IN HOSPEDING CELLS.
PCT/ES2001/000078 WO2001064914A1 (en) 2000-03-03 2001-03-02 Method for producing a recombinant alpha-galactosidase of thermus sp. t2 in host cells
AU37444/01A AU3744401A (en) 2000-03-03 2001-03-02 Method for producing a recombinant alpha-galactosidase of thermus sp. t2 in hostcells

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
ES200000515A ES2172380B1 (en) 2000-03-03 2000-03-03 A PROCEDURE TO PRODUCE ALFA-GALACTOSIDASA RECOMBINANT THERMUS SP. T2 IN HOSPEDING CELLS.

Publications (2)

Publication Number Publication Date
ES2172380A1 true ES2172380A1 (en) 2002-09-16
ES2172380B1 ES2172380B1 (en) 2003-12-01

Family

ID=8492553

Family Applications (1)

Application Number Title Priority Date Filing Date
ES200000515A Expired - Fee Related ES2172380B1 (en) 2000-03-03 2000-03-03 A PROCEDURE TO PRODUCE ALFA-GALACTOSIDASA RECOMBINANT THERMUS SP. T2 IN HOSPEDING CELLS.

Country Status (3)

Country Link
AU (1) AU3744401A (en)
ES (1) ES2172380B1 (en)
WO (1) WO2001064914A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010116020A1 (en) 2009-04-08 2010-10-14 Universidade Da Coruña Strains of s.cerevisiae, capable of growing in media with melibiose, stachyose and raffinose

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108359655B (en) * 2018-02-08 2020-04-24 刘丹妮 Lipase mutant TDL-mut with high thermal stability and coding gene thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2137115A1 (en) * 1997-08-07 1999-12-01 Consejo Superior Investigacion A procedure for producing recombinant [beta]-galactosidase from Thermus sp. (strain T2) in host cells, and for purifying it in a single chromatographic step.

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2137115A1 (en) * 1997-08-07 1999-12-01 Consejo Superior Investigacion A procedure for producing recombinant [beta]-galactosidase from Thermus sp. (strain T2) in host cells, and for purifying it in a single chromatographic step.

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
BERGER, J.-L. et al. "Identification of new enzyme activities of several strains of Thermus species", APPL. MICROBIOL. BIOTECHNOL., 1995, Vol. 44, paginas 81-87. Todo el documento. *
OLAFUR FRIDJONSSON et al. "Cloning of the gene encoding a novel thermostable alpha-Galactosidase from Thermus brockianus ITI360", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1999, Vol. 65, No 9, paginas 3955-3963. Todo el documento. *
OLAFUR FRIDJONSSON et al. "The structure of the alpha-galactosidase gene loci in Thermus brockianus ITI360 and Thermus thermophilus TH125", EXTREMOPHILES, 2000, Vol. 4, paginas 23-33. Todo el documento. *
VIAN, A. et al. "Structure of the beta-Galactosidase gene from Thermus sp. strain T2: Expression in Escherichia coli and purification in a single step of an active fusion protein", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1998, Vol. 64, No 6, paginas 2187-2191. Todo el documento. *
YOSHINORI KOYAMA et al. "Cloning of alpha and beta-Galactosidase genes from an extreme thermophile, Thermus strain T2, and their expression in Thermus thermophilus HB27", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1990, Vol. 56, No 7, paginas 2251-2254. Todo el documento. *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010116020A1 (en) 2009-04-08 2010-10-14 Universidade Da Coruña Strains of s.cerevisiae, capable of growing in media with melibiose, stachyose and raffinose

Also Published As

Publication number Publication date
WO2001064914A1 (en) 2001-09-07
AU3744401A (en) 2001-09-12
ES2172380B1 (en) 2003-12-01

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