Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing oxygen-containing organic compound
Reexamination Certificate
2001-01-18
2003-08-05
Lankford, Jr., Leon B. (Department: 1651)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing oxygen-containing organic compound
C435S135000, C435S136000, C514S549000, C514S551000, C514S560000
Reexamination Certificate
active
06602690
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a process for the production of dihomo-&ggr;-linolenic acid (DGLA) and a lipid containing DGLA by fermentation using a microorganism capable of producing arachidonic acid (ARA) and having reduced or lost &Dgr;5 desaturation activity.
2. Related Art
DGLA, also known as 8,11,14-eicosatrienoic acid, is known to be present as a constituent fatty acid of fish oils and seaweeds. However, because of the low content of DGLA in fish oils and the like, a purified DGLA product obtained from fish oils and seaweeds is expensive. As relatively efficient production processes, there are fermentation methods using a microorganism capable of producing DGLA (Japanese Unexamined Patent Publication No. 63-14696), methods providing enhanced productivity by adding an additive, such as an unsaturated fatty acid (Japanese Unexamined Patent Publication Nos. 64-47384, and 64-47385), and methods using sesame oil, one of various plant extracts, sesamin, episesamin or the like (Japanese Unexamined Patent Publication Nos. 1-243992, 2-268690, 3-72892 and 3-49688).
Various studies have been carried out regarding the actions of essential fatty acids on an organism, and it is known that in many cases eicosanoids derived from DGLA and ARA are antagonistic. Although it is known that a group of prostaglandin 1 group derived from DGLA exhibit platelet anti-coagulation action, vasodilation activity, bronchdilation activity anti-inflammatory action, and the like, for DGLA orally taken as a fat or oil in foods so as to exhibit the above-mentioned actions, DGLA-containing oil or fat product that has a low content of ARA antagonistic to DGLA is most preferable. Moreover, in the case wherein DGLA is purified from a DGLA-containing fat or oil, a DGLA-containing fat or oil having a low content of ARA is preferable as a starting material. Thus, the development of DGLA-containing fat or oil having a low content of ARA is in urgent demand, but a process for the production of such a fat or oil is not known.
SUMMARY OF THE INVENTION
Accordingly, the present invention provides a process for the production of DGLA and a lipid containing DGLA in a simple and efficient manner using a conventional inexpensive medium, and a process for the production of a lipid containing DGLA having a low content of ARA by adding an additive such as a &Dgr;5 desaturase inhibitor.
The present invention have found that DGLA, which is a precursor of ARA, accumulates in a large amount when a microorganism capable of producing ARA and having reduced or lost &Dgr;5 desaturation activity is cultured in a conventional medium, and that a ratio of ARA is further decreased and a ratio of DGLA is further increased when said microorganism is cultured in a medium supplemented with a &Dgr;5 desaturation inhibitor.
Accordingly, the present invention provides a process for the production of DGLA comprising the steps of:
culturing a microorganism having an ability to produce ARA and having reduced or lost &Dgr;5 desaturation activity in a medium to produce DGLA or a lipid containing DGLA; and
recovering the DGLA.
The present invention also provides a process for the production of a lipid containing DGLA comprising the steps of:
culturing a microorganism having an ability to produce ARA and having reduced or lost &Dgr;5 desaturation activity in a medium to produce a lipid containing DGLA; and
recovering the lipid containing DGLA.
The present invention further provides a process for the production of DGLA comprising the steps of:
culturing a microorganism having an ability to produce ARA and having reduced or lost &Dgr;5 desaturation activity in a medium supplemented with a &Dgr;5 desaturase inhibitor, or adding a &Dgr;5 desaturase inhibitor into a medium in which said microorganism has been cultured and further culturing the microorganism to produce DGLA or a lipid containing DGLA; and
recovering the DGLA.
The present invention further provides a process for the production of a lipid containing DGLA comprising the steps of:
culturing a microorganism having an ability to produce ARA and having reduced or lost &Dgr;5 desaturation activity in a medium supplemented with a &Dgr;5 desaturase inhibitor, or adding a &Dgr;5 desaturase inhibitor into a medium in which said microorganism has been cultured and further culturing the microorganism to produce a lipid containing DGLA; and
recovering the lipid containing DGLA.
The present invention moreover provides a process for the production of DGLA comprising the steps of:
culturing a microorganism having an ability to produce ARA and having reduced or lost &Dgr;5 desaturation activity in a medium supplemented with at least one additive selected from the group consisting of sesame oil, peanut oil, an extract obtained by extracting sesame oil with an organic solvent substantially immiscible with sesame oil, an extract of sesame seeds, an extract of Gokahi derived from a medicinal plant, which is
Acanthopanax gracilistylus
W. W. Smith,
Acanthopanax senticosus
Harms,
Acanthopanax henryi
, or
Acanthopanax verticillatus
Hoo, an extract of Touboku derived from a medicinal plant, an which is
Paulownia fortunei
Hemsl; or
Paulownia tomentosa
Steud., an extract of Hakukajihi derived from a medicinal plant, an which is of
Ginkgo biloba
L., an extract of Hihatsu derived from a medicinal plant, an extract of
Piper longum
L., which is Saishin (
Asiasari radix
) derived from medicinal plant, a which is
Asiasarum heterotropoides
var. mandshuricum, an extract of
Asarum sieboldii
Miq., an extract of tarragon, an extract of dill seeds, an extract of parsley, an extract of turmeric and an extract of nutmeg, or adding said additive into a medium in which said microorganism has been cultured and further culturing the microorganism to produce DGLA or a lipid containing DGLA, and
recovering the DGLA.
The present invention further provides a process for the production of a lipid containing DGLA comprising the steps of:
culturing a microorganism having an ability to produce ARA and having reduced or lost &Dgr;5 desaturation activity in a medium supplemented with at least one additive selected from the group consisting of sesame oil, peanut oil, an extract obtained by extracting sesame oil with an organic solvent substantially immiscible with sesame oil, an extract of sesame seeds, an extract of Gokahi derived from a medicinal plant, an extract of
Acanthopanax gracilistylus
W. W. Smith, an extract of
Acanthopanax senticosus
Harms, an extract of
Acanthopanax henryi,
an extract of
Acanthopanax verticillatus
Hoo, an extract of Touboku derived from a medicinal plant, an extract of
Paulownia fortunei
Hemsl; an extract of
Paulownia tomentosa
Steud., an extract of Hakukajihi derived from a medicinal plant, an extract of
Ginkgo biloba
L., an extract of Hihatsu derived from a medicinal plant, an extract of
Piper longum
L., an extract of Saishin (
Asiasari radix
) derived from medicinal plant, an extract of
Asiasarum heterotropoides
var. mandshuricum, an extract of
Asarum sieboldii
Miq., an extract of tarragon, an extract of dill seeds, an extract of parsley, an extract of turmeric and an extract of nutmeg, or adding said additive into a medium in which said microorganism has been cultured and further culturing the microorganism to produce a lipid containing DGLA, and
recovering the lipid containing DGLA.
DETAILED DESCRIPTION
In the present invention any microorganisms having an ability to produce ARA and having reduced or lost &Dgr;5 desaturase activity can be used. Microorganism having an ability to produce ARA include those belonging to the genus Mortierella, Conidiobolus, Pythium, Phytophthora, Penicillium, Cladosporium, Mucor, Fusarium, Aspergillus, Rhodotorula, or Entomophthora. As microorganisms belonging to the genus Mortierella, there are mentioned microorganisms belonging to the subgenus Mortierella, such as
Mortierella elongata, Mortierella exiqua, Mortierella hyprophila, Mortierella alpina,
and the like. Microorganism used in th
Akimoto Kengo
Kawashima Hiroshi
Shimizu Sakayu
Yamada Hideaki
Burns Doane Swecker & Mathis L.L.P.
Lankford , Jr. Leon B.
Suntory Ltd.
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