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ES2172380B1 - 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
ES2172380B1
ES2172380B1 ES200000515A ES200000515A ES2172380B1 ES 2172380 B1 ES2172380 B1 ES 2172380B1 ES 200000515 A ES200000515 A ES 200000515A ES 200000515 A ES200000515 A ES 200000515A ES 2172380 B1 ES2172380 B1 ES 2172380B1
Authority
ES
Spain
Prior art keywords
alpha
thermus
cells
galactosidase
hospeding
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.)
Expired - Fee Related
Application number
ES200000515A
Other languages
Spanish (es)
Other versions
ES2172380A1 (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)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes 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 process 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 thermostable alpha-galactosidase (EC 3.2.1.22). The way to obtain a genetic construct for the expression of said nucleotide sequence in Escherichia coli cells is indicated by cloning it into an expression vector, with the corresponding modifications in the nucleotide and amino acid sequences to which manipulation leads. A method for obtaining enriched extracts with thermostable alpha-galactosidase activity produced in E. coli is described. The described recombinant protein, due to its thermostability characteristic, which makes it compatible with various technological procedures, has application mainly in the agri-food-sanitary sector, for the hydrolysis of poorly digestible alpha-galactosides present in various high-consumption 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 ES2172380A1 (en) 2002-09-16
ES2172380B1 true 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)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2351296B8 (en) 2009-04-08 2012-07-03 Universidade Da Coruña CEPAS DE S. CEREVISIAE CAPACIES OF GROWING IN MEDIA WITH MELIBIOUS, STAQUIOUS AND RAFINOSA.
CN108359655B (en) * 2018-02-08 2020-04-24 刘丹妮 Lipase mutant TDL-mut with high thermal stability and coding gene thereof

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2137115B1 (en) * 1997-08-07 2000-08-16 Consejo Superior Investigacion A PROCEDURE FOR PRODUCING RECOMBINANT BETA-GALACTOSIDASE FROM THERMUS SP. (CEPA T2) IN HOSPEDANT CELLS, AND PURIFY IT IN A SINGLE CHROMATOGRAPHIC STEP.

Also Published As

Publication number Publication date
WO2001064914A1 (en) 2001-09-07
ES2172380A1 (en) 2002-09-16
AU3744401A (en) 2001-09-12

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