EP2590732A1 - Membran mit konstitutiv offenen aquaporinen - Google Patents

Membran mit konstitutiv offenen aquaporinen

Info

Publication number
EP2590732A1
EP2590732A1 EP11733640.4A EP11733640A EP2590732A1 EP 2590732 A1 EP2590732 A1 EP 2590732A1 EP 11733640 A EP11733640 A EP 11733640A EP 2590732 A1 EP2590732 A1 EP 2590732A1
Authority
EP
European Patent Office
Prior art keywords
seq
membrane
aquaporin
water
aquaporins
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.)
Withdrawn
Application number
EP11733640.4A
Other languages
English (en)
French (fr)
Inventor
Per Kjellbom
Urban Johansson
Andreas Kirscht
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.)
Hydrogene Lund AB
Original Assignee
Hydrogene Lund AB
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 Hydrogene Lund AB filed Critical Hydrogene Lund AB
Publication of EP2590732A1 publication Critical patent/EP2590732A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
    • B01D69/14—Dynamic membranes
    • B01D69/141—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes
    • B01D69/142—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes with "carriers"
    • B01D69/144—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes with "carriers" containing embedded or bound biomolecules
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F1/00—Treatment of water, waste water, or sewage
    • C02F1/44—Treatment of water, waste water, or sewage by dialysis, osmosis or reverse osmosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants

Definitions

  • the present invention relates to a constitutively open aquaporin. Further, it relates to a membrane comprising such an aquaporin and a use of such membrane to purify water. The present invention also relates to a method of converting aquaporins, to constitutively open aquaporins.
  • Naturally occuring biological membranes have only limited intrinsic water permeability cells maintain the flux of water into and out of the cell via a family of water-specific, membrane protein channels called aquaporins.
  • aquaporin family are well known in literature, and found in archea, eubacteria and eukaryotes, including fungi, animals and plants. They serve an astonishing variety of physiological functions and are easily identified by sequence similarity across all kingdoms of life. In higher eukaryotes, water transport activity of aquaporins is frequently regulated by phosphorylation, pH and osmolarity. Aquaporins in plants and animals are highly conserved and form large protein families with 35 members in higher plants and 13 members in humans.
  • plant aquaporins are further divided into four subfamilies and their presence in primitive plants such as the bryophyte
  • PIPs plasma membrane aquaporins
  • the present invention preferably seeks to mitigate, alleviate or eliminate one or more of the above-identified deficiencies in the art and disadvantages singly or in any combination and solves at least the above mentioned problems by providing a device, a method and a use, according to the appended patent claims.
  • the general solution according to the invention is to utilize constitutively open aquaporins in a synthetic membrane.
  • a membrane comprising a constitutively open His-tagged aquaporin is provided.
  • the constitutively open His-tagged aquaporin may have at least 90 %, such as at least 95, 97 or 99%, sequence homology with SEQ ID NO: 2.
  • the membrane may comprise a constitutively open His-tagged aquaporin having amino acid sequence according to SEQ ID NO: 2.
  • the aquaporin of the membrane originates from Spinacia oleracea, Arabidopsis thaliana, Zea mays, Oryza sativa or Physcomitrella patens.
  • the aquaporin may comprise a sequence having at least 90 %, such as at least 95, 97 or 99%, sequence homology with any of SEQ ID NO: 1, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, or SEQ ID NO: 30.
  • the aquaporin may comprise a C-terminal His-tag having at least 90 %, such as at least 95, 97 or 99%>, sequence homology with SEQ ID NO: 3.
  • the aquaporin comprises the sequence according to SEQ ID NO: 1
  • SEQ ID NO: 1 SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, or SEQ ID NO: 30, and a C-terminal His-tag having according to SEQ ID NO: 3.
  • a device for purifying water comprises an inlet connected to a first chamber, for receiving dirty water, and an outlet connected to a second chamber for dispensing clean water, and a water purifying membrane comprising the membrane according to according to the first aspect, arranged in between the first and the second chamber.
  • a third aspect of the invention use of the membrane according to the first aspect or a device according to the second aspect, for purifying water is provided.
  • a method for producing a membrane according to the first aspect comprising the steps of: producing a His-tagged aquaporin, which is constitutively open; and incorporating the aquaporin into a membrane.
  • the present invention has the advantage over the prior art that it offers increased water permeability without negatively affecting the water purification.
  • Fig. 1 is a graph according to an embodiment of the invention.
  • Aquaporins e.g. SoPIP2;l (SEQ ID NO: 1)
  • SoPIP2;l SEQ ID NO: 1
  • SoPIP2;l (SEQ ID NO: 1) belong to one of the best characterized classes of aquaporins, the PIPs.
  • PIPs are known to be gated by phosphorylation of serine amino acids causing the channel to open and by protonation of a histidine amino acid or by binding of Ca 2+ causing the channel to close (Tornroth-Horsefield et al. Nature 439, 688-694, 2006).
  • AtTIP2;l is water permeable when reconstituted in proteo liposomes, but quantitative comparison of the water transport indicated that the permeability of AtTIP2;l is approximately two thirds of the permeability of SoPIP2;l (SEQ ID NO: 1). In contrast the thermal stability of AtTIP2;l is much higher, with a melting point of 86° C compared to 57° C for So-PIP2; 1.
  • Aquaporin proteins are functioning as water-specific channels in cell membranes. Genes encoding aquaporin proteins have been inserted in a specific yeast strain and the aquaporin protein produced and purified in its functional form and used for constructing the water- specific membranes.
  • the gene coding for the spinach plasma membrane aquaporin SoPIP2;l (SEQ ID NO: 1) has been cloned into the yeast Pichia pastoris, overexpressed and purified. Without buffer exchange the yield is about 20-30 mg pure SoPIP2;l per preparation.
  • the constitutively open conformation of aquaporin is achieved when the C-terminal His-tag prevents interaction between Ser274 of aquaporin, and the next monomer which can be seen in a closed conformation, depending on the exact type of aquaporin.
  • Ser274 is however conserved in all aquaporins belonging to the PIP2 group, both in higher plants and more primitive plants.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Botany (AREA)
  • Dispersion Chemistry (AREA)
  • Hydrology & Water Resources (AREA)
  • Engineering & Computer Science (AREA)
  • Environmental & Geological Engineering (AREA)
  • Water Supply & Treatment (AREA)
  • Peptides Or Proteins (AREA)
  • Separation Using Semi-Permeable Membranes (AREA)
EP11733640.4A 2010-07-08 2011-07-07 Membran mit konstitutiv offenen aquaporinen Withdrawn EP2590732A1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE1050751 2010-07-08
PCT/EP2011/061539 WO2012004357A1 (en) 2010-07-08 2011-07-07 Membrane comprising constitutively open aquaporins

Publications (1)

Publication Number Publication Date
EP2590732A1 true EP2590732A1 (de) 2013-05-15

Family

ID=44583555

Family Applications (1)

Application Number Title Priority Date Filing Date
EP11733640.4A Withdrawn EP2590732A1 (de) 2010-07-08 2011-07-07 Membran mit konstitutiv offenen aquaporinen

Country Status (5)

Country Link
EP (1) EP2590732A1 (de)
JP (1) JP5912113B2 (de)
DK (1) DK2590732T1 (de)
SG (2) SG10201505266QA (de)
WO (1) WO2012004357A1 (de)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090007555A1 (en) 2005-09-20 2009-01-08 Peter Holme Jensen Biomimetic Water Membrane Comprising Aquaporins Used in the Production of Salinity Power
EP1885477B1 (de) 2005-05-20 2010-02-17 Aquaporin APS Membran zum filtrieren von wasser

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20070114316A (ko) * 2002-07-29 2007-11-30 엠티 테크날러지스, 인코포레이션 생체유사 막
WO2009076174A1 (en) * 2007-12-05 2009-06-18 The Board Of Trustees Of The University Of Illinois Highly permeable polymer membranes

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1885477B1 (de) 2005-05-20 2010-02-17 Aquaporin APS Membran zum filtrieren von wasser
US7857978B2 (en) 2005-05-20 2010-12-28 Aquaporin A/S Membrane for filtering of water
US20090007555A1 (en) 2005-09-20 2009-01-08 Peter Holme Jensen Biomimetic Water Membrane Comprising Aquaporins Used in the Production of Salinity Power

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO2012004357A1

Also Published As

Publication number Publication date
JP5912113B2 (ja) 2016-04-27
WO2012004357A1 (en) 2012-01-12
JP2013535312A (ja) 2013-09-12
SG186897A1 (en) 2013-02-28
DK2590732T1 (da) 2015-06-22
SG10201505266QA (en) 2015-08-28

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