Food or edible material: processes – compositions – and products – Fermentation processes
Reexamination Certificate
2001-02-19
2003-04-01
Lilling, Herbert J. (Department: 1651)
Food or edible material: processes, compositions, and products
Fermentation processes
C426S042000, C426S043000, C426S060000, C426S061000, C426S491000, C426S492000, C426S580000, C426S585000, C426S581000, C426S601000
Reexamination Certificate
active
06541049
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to a process for production of arachidonic acid. The present invention also relates to a food product and a method to make the food product containing such arachidonic acid.
BACKGROUND OF THE INVENTION
Arachidonic acid (all-cis-5,8,11,14-eicosatetraenoic acid) is a polyunsaturated fatty acid (PUFA) containing 20 carbon atoms with four double bonds. The double bonds are arranged with the last one located six carbon atoms from the methyl end of the chain. Therefore, arachidonic acid is referred to as an omega-6 fatty acid. Arachidonic acid is one of the most abundant C
20
PUFA's in the human body. It is particularly prevalent in organ, muscle and blood tissues. Arachidonic acid is a direct precursor for a number of circulating eicosenoids, such as prostaglandins, thromboxanes, leukotrienes and prostacyclins, which are important biological regulators. These eicosenoids exhibit regulatory effects on lipoprotein metabolism, blood rheology, vascular tone, leukocyte function, platelet activation and cell growth. The application of arachidonic acid to an infant's diet is particularly important due to the rapid body growth of an infant. Arachidonic acid is an important precursor to many of the eicosanoids which regulate cellular metabolism and growth in infants. It is found naturally in human breast milk but not in most infant formula. In an effort to have infant formula match the long chain fatty acid profile found in breast milk, scientific and food regulatory bodies have recommended that arachidonic acid be added to infant formula, especially in formula utilized for premature infants.
In particular, it is preferable that arachidonic acid containing oil produced for use with infant formula contain little or no other long chain highly unsaturated fatty acids (e.g., eicosapentanoic acid). Such other long chain highly unsaturated fatty acids are not preferred because some of these fatty acids can interfere with the utilization of arachidonic acid by the infant, and/or can inhibit blending of the arachidonic acid-containing oil with other oils to achieve the appropriate ratio of fatty acids matching breast milk or other desired applications. Highly unsaturated fatty acids are defined as fatty acids containing 4 or more double bonds.
Traditional sources of arachidonic acid include poultry eggs, bovine brain tissue, pig adrenal gland, pig liver and sardines. The yield of arachidonic acid, however, is usually less than 0.2% on a dry weight basis. The use of microorganisms capable of producing arachidonic acid de novo have been suggested by various investigators, including Kyle, PCT Publication No. WO 92/13086, published Aug. 6, 1992; Shinmen et al., U.S. Pat. No. 5,204,250, issued Apr. 20, 1993; Shinmen et al., pp. 11-16, 1989,
Appl. Microbiol. Biotechnol.,
vol. 31; Totani et al., pp. 1060-1062, 1987,
LIPIDS,
vol. 22; Shimizu et al., pp. 509-512, 1992,
LIPIDS,
vol. 27; Shimizu et al., pp. 342-347, 1989,
JAOCS,
vol. 66; Shimizu et al., pp. 1455-1459, 1988,
JAOCS,
vol. 65; Shimizu et al., pp. 254-258, 1991,
JAOCS,
vol. 68; Sajbidor et al., pp. 455-456, 1990,
Biotechnology Letters,
vol. 12; Bajpai et al., pp. 1255-1258, 1991,
Appl. Environ. Microbiol.,
vol. 57; Bajpai, pp. 775-780, 1991,
JAOCS,
vol. 68; and Gandhi et al., pp. 1825-1830, 1991,
J. Gen. Microbiol.,
vol. 137. The arachidonic acid productivity by the microorganisms disclosed by prior investigators, however, is less than 0.67 grams per liter per day. Such amounts are significantly less than the amounts of arachidonic acid produced by the microorganisms of the present invention. These lower productivity values are the result of employing strains: (1) with slow growth or lipid production rates leading to long fermentation times (i.e., greater than 2-3 days) (Kyle, 1992, ibid.; Shinmen et al., 1993, ibid.; Shinmen et al., 1989, ibid.; Bajpai et al., 1991, ibid.; Bajpai, ibid.; and Gandhi et al., ibid.); and/or (2) that contain low arachidonic acid contents (expressed as % fatty acids) in the final oil produced (Shinmen et al., 1993, ibid.; Shimizu et al., 1989, ibid.; and Kendrick and Ratledge, 1992, pp. 15-20,
Lipids,
vol. 27); and/or (3) which require long periods of stress (i.e., aging a biomass for 6-28 days) to achieve high levels of arachidonic acid in a biomass (Bajpai et al., 1991, ibid. and Shinmen et al., 1989, ibid.); and/or (4) that only exhibit high arachidonic acid content in non-commercial growth conditions (e.g., malt agar plates) (Totani and Oba, 1987, pp. 1060-1062,
Lipids,
vol. 22). In addition, non-
Mortierella schmuckeri
microorganisms that have been proposed for producing arachidonic acid, in particular
Pythium insidiosum
microorganisms, disclosed by prior investigators (Kyle, 1992, ibid.), have been reported to be pathogenic to humans and/or animals.
Thus, there remains a need for an economical, commercially feasible method for producing arachidonic acid. The present invention satisfies that need. There also remains a need for the an economical, commercially feasible food product for the introduction of arachidonic acid produced according to the present invention into the diet of human infants.
SUMMARY
The present invention provides for a method for economically producing arachidonic acid. One embodiment of the present invention includes a method to produce arachidonic acid, comprising culturing microorganisms of the genus
Mortierella
sect.
schmuckeri
in a medium comprising a source of assimilable organic carbon and a source of assimilable nitrogen. In another embodiment, such strains of
Mortierella
sect.
schmuckeri
are capable of producing at least about 0.86 grams per liter per day of arachidonic acid.
Yet another embodiment of the present invention includes a food product comprising lipids recovered from a microorganism of the genus
Mortierella
sect.
schmuckeri
and a food material. In particular, such lipids can be added to infant formula and baby food to increase the arachidonic acid or long chain omega-6 fatty acid content of such foods.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides for a novel process for the production of commercially feasible amounts of arachidonic acid using a
Mortierella
sect.
schmuckeri
microorganism. One embodiment of the present process is to produce arachidonic acid by culturing microorganisms of the genus
Mortierella
sect.
schmuckeri
in a medium comprising a source of assimilable organic carbon and a source of assimilable nitrogen. The lower fungi Phycomycetes contains at least two classes, including Oomycetes and Zygomycetes. The class Zygomycetes contains at least two orders, including Entomophthorales and Mucorales. Contained within the Mucorales order are numerous genera including Mortierella. The genus Mortierella contains nine sections, including sect.
schmuckeri
(Gams, 1977, pp. 381-391,
Persoonia,
vol. 9 and Gams, 1977, p. 216, in Abstracts Vol. A-L, Second International Mycological Congress, University of South Florida). The
schmuckeri
sect. of the genus Mortierella contains three species referred to as
Mortierella camargensis, Mortierella clausenii
and
Mortierella schmuckeri.
All of the other strains of Mortierella that have been evaluated for arachidonic acid production belong to the
Mortierella
sections
alpina, hygrophila
or
spinosa.
It has now been recognized that strains of
Mortierella
sect.
schmuckeri
are particularly advantageous in the production of arachidonic acid compared to these other strains of Mortierella. In particular, it has been found that strains of
Mortierella
sect.
schmuckeri
are capable of producing arachidonic acid with high productivity. Strains of
Mortierella
sect.
schmuckeri
of the present invention are preferably capable of producing at least about 0.70 grams of arachidonic acid per liter per day, more preferably at least about 0.80 grams of arachidonic acid per liter per day, and even more preferably at least about 0.86 grams of arachidonic acid per liter per day. Preferably, strains
Lilling Herbert J.
Martek Biosciences Boulder Corporation
Sheridan & Ross P.C.
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