AU2012232064B2 - Method of producing lauric acid-containing oil or fat and lauric acid or esters thereof - Google Patents

Abstract

To provide a method for supplying oil or fat containing lauric acid as a constituent fatty acid and a method for supplying lauric acid or esters thereof using the oil or fat with algae. A method for producing lauric acid or esters thereof, which method includes, culturing at least one species selected from the group consisting of algae in the class Chlorarachniophyceae and algae in the class Cryptophyceae consisting of algae belonging to the genus Rhodomonas and algae belonging to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and Chroomonas placoidea in a medium; recovering, from the culture product, an oil or fat having a lauric acid content of 3 weight% or higher of the fatty acid composition; and, as needed, esterifying the lauric acid in the recovered oil or fat, followed by separating and recovering the lauric acid or esters thereof.

Description

WO 2012/128396 PCT/JP20 12/058505 DESCRIPTION METHOD OF PRODUCING LAURIC ACID-CONTAINING OIL OR FAT AND LAURIC ACID OR ESTERS THEREOF [Field of the Invention] (0001] The present invention relates to a method for producing an oil or fat containing lauric acid as a constituent fatty acid (hereinafter may also be referred to simply as "lauric acid-containing oil or fat") and lauric acid or esters thereof using the oil or fat, the method employing algae. [Background of the Invention] [0002] Lauric acid is a typical fatty acid contained in a large amount in coconut oil and palm kernel oil and is used as a raw material of a variety of surfactants, in foods, and for other materials. Currently, the supply source of lauric acid is limited to coconut and palm kernels, which are grown in limited areas in the world. Cultivated lands now allocated to production of such lauric acid sources will be shared competitively with areas for bio-fuel for diesel engines and for food production. Excessive land cultivation for the production of lauric acid sources causes destruction of tropical rain forests. Therefore, there is demand for creating a technique for 1 supplying lauric acid, which technique does not rely on coconut or palm kernels. [ 00031 5 Meanwhile, algae are disclosed to effectively produce an oil or fat, and the productivity per area of the algae is about 10 times that of a plant or the like (Non-Patent Document 1). Among algae, dinophyceae Crypthecodinium chonii, which grows not via photosynthesis but via heterotrophy, is disclosed to be 10 a lauric acid-producing organism and to have high lauric acid content (15.7%/total lipid) (Non-Patent Document 2). [ 00041 From the viewpoints of cost for carbon sources and other factors, more preferred are algae species which can grow via 15 photosynthesis (autotrophy) and have higher lauric acid content. However, among such photoautotrophic algae species, only Neochloris oleoabundans, having a lauric acid content of about 1 to 2% at best, is disclosed (Non-Patent Document 3), and no algae species has heretofore been disclosed to have 20 higher lauric acid content. [Non-Patent Document 1]: Biotechnology Advances, (2007) 25, 294-306 [Non-Patent Document 2]: Phytochemistry, (1988) 27, 1679-1683 [Non-Patent Document 3]: J. Ind. Microbiol. Biotechnol. (2009) 25 36: 821-826 [ 0004a] The discussion of documents, acts, materials, devices, articles and the like is included in this specification solely for the purpose of providing a context for the present 30 invention. It is not suggested or represented that any or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application. 2 [ 0004b] Where the terms "comprise", "comprises", "comprised" or "comprising" are used in this specification (including the claims) they are to be interpreted as specifying the presence 5 of the stated features, integers, steps or components, but not precluding the presence of one or more other features, integers, steps or components, or group thereof. [Summary of the Invention] 10 [ 00051 In one aspect, the present invention provides a method for producing lauric acid or esters thereof, which method comprises, Culturing, in a medium, at least one species selected from 15 the group consisting of algae in the class Chlorarachniophyceae and algae in the class Cryptophyceae consisting of algae belonging to the genus Rhodomonas and algae belonging to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and 20 Chroomonas placoidea, Wherein the algae in the class Chlorarachniophyceae is Lotherella globosa strain CCMP1729 or an algae strain having virtually the same phycological properties as those of CCMP1729; 25 recovering, from the cultured algae, an oil or fat having a lauric acid content of 3 weight% or higher of the fatty acid composition; and, as needed, esterifying the lauric acid in the recovered oil or fat, followed by separating and recovering the lauric acid or esters thereof. 30 [ 0005al In another aspect, the present invention provides a method for producing an oil or fat comprising lauric acid as a constituent fatty acid, which method comprises culturing, in a medium, at least one species of algae belonging to the genus 3 Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and Chroomonas placoidea and recovering, from the cultured algae, an oil or fat comprising a lauric acid content of 3 weight% or 5 higher of the fatty acid composition. [ 0005b] In a further aspect, the present invention provides a method for producing lauric acid or esters thereof, which method including: culturing at least one species selected from 10 the group consisting of algae in the class Chlorarachniophyceae and algae in the class Cryptophyceae consisting of algae belonging to the genus Rhodomonas and algae belonging to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and 15 Chroomonas placoidea in a medium; recovering, from the culture product, an oil or fat having a lauric acid content of 3 weight% or higher of the fatty acid composition; and, as needed, esterifying the lauric acid in the recovered oil or fat, followed by separating and recovering the lauric acid or 20 esters thereof. Further, the present invention provides a method for producing an oil or fat containing lauric acid as a constituent fatty acid, which method including: culturing, in a medium, at least one species of algae belonging to the genus Chroomonas 25 selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and Chroomonas placoidea; and recovering, from the culture product, an oil or fat having a lauric acid content of 3 weight% or higher of the fatty acid composition. 30 [Modes for Carrying Out the Invention] 00061 3a WO 2012/128396 PCT/JP2012/058505 The present invention relates to provision of a method for supplying oil or fat containing lauric acid as a constituent fatty acid and a method for supplying lauric acid or esters thereof using the oil or fat, through employment of algae. [0007] The present inventors have carried out studies on lauric acid-producing organisms, and have found that among photoautotrophic algae, algae in the class of Chlorarachniophyceae which are a unicellular algae and algae belonging to the genus Rhodomonas or algae belonging to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and Chroomonas placoidea among algae in the class of Cryptophyceae have high lauric acid content, and that an oil or fat containing lauric acid as a constituent fatty acid at high content and further lauric acid or esters thereof can be efficiently produced by use of the algae. [0008] According to the method of the present invention, which employs algae that can readily grow, an oil or fat containing lauric acid as a constituent fatty acid at high content can be efficiently produced, without imposing limitation on the cultivated fields for the growth of coconut and palm kernels or competing in the cultivated land with areas for food production, etc. In addition, according to the method of the present invention, destruction of tropical rain forests can 4 WO 2012/128396 PCT/JP2012/058505 be avoided. (0009] The method of the present invention for producing a lauric acid-containing oil or fat includes culturing, in a medium, at least one species of algae in the class Cryptophyceae selected from the group consisting of algae belonging to the genus Rhodomonas and algae belonging to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and Chroomonas placoidea and recovering, from the culture product, an oil or fat having a lauric acid content of 3 weight% or higher in the fatty acid composition. Further, the method of the present invention for producing lauric acid or esters thereof includes culturing at least one species selected from the group consisting of algae in the class Chlorarachniophyceae and algae in the class Cryptophyceae consisting of algae belonging to the genus Rhodomonas and algae belonging to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and Chroomonas placoidea in a medium; recovering, from the culture product, an oil or fat having a lauric acid content of 3 weight% or higher of the fatty acid composition; and, as needed, esterifying the lauric acid in the recovered oil or fat, followed by separating and recovering the lauric acid or esters thereof. The oil or fat has a lauric acid content of 3 weight% 5 WO 2012/128396 PCT/JP2012/058505 or higher of the fatty acid composition. The lauric acid content is preferably from 3 to 60 weight%, more preferably from 5 to 60 weight%, even more preferably from 6 to 60 weight%, even more preferably from 7 to 60 weight%, even more preferably from 8 to 60 weight%, even more preferably from 9 to 60 weight%, even more preferably 10 to 60 weight%, even more preferably 11 to 5o weight%, and even more preferably 12 to 40 weight%. [0010] The algae employed in the present invention may be any algae strains in the class Chlorarachniophyceae, so long as the strains have an ability to produce an oil or fat having a lauric acid content of 3 weight% or higher in the fatty acid composition. The algae in the class Cryptophyceae employed in the present invention may be any algae strains belonging to the genus Rhodomonas or to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and Chroomonas placoidea, more preferably the algae strains belonging to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica and Chroomonas nordstedtii, so long as the strains have an ability to produce an oil or fat having a lauric acid content of 3 weight% or higher in the fatty acid composition. [0011] The algae of the present invention may be selected 6 WO 2012/128396 PCT/JP2012/058505 through, for example, the following screening procedure: i) dispensing a sterilized medium (WA medium (see Table 2) as a fresh water medium or Daigo IMK medium (see Table 3) as a seawater medium) into a culture container; ii) inoculating an alga strain to the medium and performing stationary culturing at room temperature (22 0 C to 24*C) under illumination (illuminance: about 3,000 lux, illumination for 12 hours and dark for 12 hours); iii) recovering the produced alga and extracting oil or fat; methyl esterifying the fatty acids; and determining the fatty acid composition, to thereby select an alga strain which can produce a lauric acid-containing oil or fat; and iv) selecting an alga strain having a lauric acid content of 3 weight% or higher based on the total fatty acid in the oil or fat. [00121 Examples of algae belonging to the class Chlorarachnion include algae belonging to the genus Chlorarachnion, Lotharella, Gymnochlora, Cryptochlora, and Bigelowiella. Among them, the genus Lotharella, Gymnochlora, and Bigelowiella are preferred. Examples of more preferred algae in the class Chlorarachnion include the following algae. Examples of more preferred algae belonging to the genus Lotharella include Lotharella globosa, Lotharella amoebiformis, and Lotharella vacuolata. Examples of more preferred algae belonging to the genus Gymnochlora include Gymnochlora stellata. Examples of 7 WO 2012/128396 PCT/JP2012/058505 more preferred Bigelowiella include Bigelowiella natans. Among Lotharella globosa strains, Lotharella globosa strain CCMP1729 is more preferred. Among Lotharella amoebiformis strains, Lotharella amoebiformis strain CCMP2058 is more preferred. Among Lotharella vacuolata strains, Lotharella vacuolata strain CCMP240 is preferred. Among Gymnochlora stellata strains, Gymnochlora stellata strain CCMP2057 is more preferred. Among Bigelowiella natans strains, Bigelowiella natans strains CCMP621 and CCMP2757 are more preferred (these strains are available from, for example, the Provasoli-Guillard National Center for Culture of Marine Phytoplankton (CCMP)). Strains having virtually the same phycological properties as those of algae strains are also more preferred. For example, in recent years, Lotharella amoebiformis has been proposed to be assigned to the new genus Amorphochlora amoebiformis. These strains are considered to have virtually the same phycological properties as those of Lotharella amoebiformis. Among these, Lotharella globosa strains are more preferred and Lotharella globosa strain CCMP1729 or strains having virtually the same phycological properties as the strain are even more preferred. [0013] Examples of the strain having virtually the same phycological properties as those of Lotharella amoebiformis strain CCMP2058 include Lotharella amoebiformis strain Ryukyu. Examples of the strain having virtually the same phycological properties as those of Lotharella vacuolata strain CCMP240 8 WO 2012/128396 PCT/JP2012/058505 include Lotharella vacuolata strain FK18G. Examples of the strain having virtually the same phycological properties as those of Gymnochlora stellata strain CCMP2057. include Gymnochlora stellata strain Guam-1. Examples of the strain having virtually the same phycological properties as those of Bigelowiella natans strains CCMP621 and CCMP2757 include Bigelowiella natans strains All, 490, and VA3. [0014] The aforementioned algae strains have the following phycological properties. Strains belonging to the same genus as that of the algae strains, and strains having virtually the same mycological properties as those of the algae strains can be identified on the basis of the following properties. <Phycological properties of the algae in the class Chl orarachni ophyceae> i) Containing chlorophyll a and b ii) Chloroplast surrounded by four membranes iii) Having nucleomorph iv) Not accumulating starch v) Presence of amoeba phase and cell-wall-having phase vi) Having no stigma <Phycological properties of the algae belonging to the genus Chlorarachnion> i) Amoeboid vegetative cell ii) Nucleomorph in pyrenoid <Phycological properties of the algae belonging to the genus Cryptochlora> 9 WO 2012/128396 PCT/JP2012/058505 i) Spherical vegetative cell ii) Pyrenoid structure unknown <Phycological properties of the algae belonging to the genus Lotharella> i) The pyrenoid matrix was devided into two halves by a slit of the periplastidial compartment. ii) Nucleomorph present in the periphery of chloroplast in the vicinity of the pyrenoid base <Phycological properties of the algae belonging to the genus Gymnochlora> i) Inner membranes of chloroplast envelope invaginating pyrenoid matrix ii) Nucleomorph present in the periphery of chloroplast in the vicinity of the pyrenoid base <Phycological properties of the algae belonging to the genus Bigelowiella> i) Swarmers asexually proliferating ii) The pyrenoid matrix was slightly invaded by periplastidial compartment from the tip of the pyrenoid. iii) Nucleomorph present at the pyrenoid base <Phycological properties of Lotharella globosa strain CCMP1729> i) Spherical vegetative cell, no ameba-like emerging upon proliferation <Phycological properties of Lotharella amoebiformis strain CCMP2058> i) Ameba-like vegetative cell 10 WO 2012/128396 PCT/JP2012/058505 <Phycological properties of Lotharella vacuolata strain CCMP240> i) Spherical in the main stage of life cycle, ameba like cells having filopodia observed during the initial to middle culture stage ii) Having vacuoles larger than those of the other algae belonging to the genus Lotharella iii) Vegetative proliferation through binary fission of ameba-like cells <Phycological properties of Gymnochlora stellata strain CCMP2057> i) Star-shape amebic organism having many filopodia not forming network ii) Cells having cell wall or swarmers are absent throughout the life cycle <Phycological properties of Bigelowiella natans strains CCMP621 and CCMP2757> i) Swarmers in the vegetative stage, not ameba-like cells ii) Having two flagella (short and long) iii) Having no striae [0015] As the algae belonging to the genus Chroomonas of the present invention, examples of preferred Chroomonas diplococca including Chroomonas diplococca strain UTEX LB2422; examples of preferred Chroomonas mesostigmatica including Chroomonas mesostigmatica strain NIES1370; examples 11 WO 2012/128396 PCT/JP2012/058505 of preferred Chroomonas nordstedtii including Chroomonas nordstedtii strains NIES707 and NIES710; examples of preferred Chroomonas placoidea including Chroomonas placoidea strain NIES705 (these strains are available from The culture collection of algae at University of Texas at Austin (UTEX), National Institute for Environmental Studies (NIES), etc.); and strains having virtually the same phycological properties as those of algae strains are mentioned. As the algae belonging to the genus Rhodomonas, Rhodomonas salina is preferred, with Rhodomonas salina UTEX1375, Rhodomonas salina CCMP272, and strains having virtually the same phycological properties as those of algae strains being more preferred; and Rhodomonas salina UTEX1375 or strains having virtually the same phycological properties as those of algae strains being even more preferred. These strains are available from UTEX and The Provas.oli-Guillard National Center for Culture of Marine Phytoplankton (CCMP). [0016] Examples of the strain having virtually the same phycological properties as those of Chroomonas mesostigmatica strain NIES1370 include Chroomonas mesostigmatica strain TKB 112. Examples of the strain having virtually the same phycological properties as those of Chroomonas nordstedtii strain NIES707 include Chroomonas nordstedtii strain #00173. Examples of the strain having virtually the same phycological properties as those of Chroomonas nordstedtii strain NIES710 include Chroomonas nordstedtii strain #00331. Examples of 12 WO 2012/128396 PCT/JP2012/058505 the strain having virtually the same phycological properties as those of Chroomonas placoidea strain NIES705 include Chroomonas placoidea strain CCAP 978/8. Examples of the strain having virtually the same phycological properties as those of Rhodomonas salina strain CCMP272 include Rhodomonas salina strain Mel-023. [00171 The aforementioned algae strains have the following phycological properties. Strains belonging to the same genus as that of the algae strains, and strains having virtually the same mycological properties as those of the algae strains can be identified on the basis of the following properties. <Phycological properties of the class Cryptophyceae> i) Containing phycobilin and chlorophyll c ii) Chloroplast surrounded by four membranes iii) Having nucleomorph iv) Having tubular pleuronematic and tubular unilateral flagella v) Accumulating tx-1,4-starch <Phycological properties of the algae belonging to the genus Chroomonas> i) Barrel-form cell with no cingulum ii) Having two ejectisomes in a row iii) Blue to green chloroplast iv) Stigma generally observed centrally in a cell <Phycological properties of the algae belonging to the genus Rhodomonas> 13 i) Egg-form cell having a short cingulum ii) Red to reddish brown chloroplast with distinct pyrenoid iii) Stigma generally observed centrally in a cell 5 <Phycological properties of Chroomonas mesostigmatica strain NIES1370> i) Having a large number of lamellar structures in a chloroplast ii) Having one large pyrenoid with a chloroplast 10 <Phycological properties of Chroomonas nordstedtii strains NIES707 and NIES710> i) Having no stigma ii) Having phototaxis with respect to light having a wavelength of 450 nm to 650 nm 15 <Phycological properties of Chroomonas placoidea strain NIES705> i) Having ligules at a flagellum bearing <Phycological properties of Rhodomonas salina UTEX1375 and Rhodomonas salina CCMP272> 20 i) Two flagellua shorter than the cell length arising from a subapical end of the cell ii) Short sulcus, gullet with ejectisomes in two rows, reaching to the cell center iii) Having one reddish brown to yellowish orange 25 chloroplast, with one pyrenoid being dorsal and surrounded by marked starch sheath 00181 14 WO 2012/128396 PCT/JP2012/058505 The algae of the present invention also encompass mutants of the aforementioned algae strains and strains having virtually the same mycological properties as those of the aforementioned algae strains. For example, a mutant strain designed so as to produce an oil or fat having a higher lauric acid content as compared with a corresponding wild-type strain is also included in the algae of the present invention. Furthermore, a gene derived from the algae in the class Chlorarachniophyceae and a gene derived from the algae in the class Cryptophyceae may be employed to produce an oil or fat having a high lauric acid content. [0019] The algae in the class Chlorarachniophyceae and the algae in the class Cryptophyceae of the present invention may be cultured in an appropriate medium prepared from natural or artificial seawater under illumination through a cultivation method generally employed in culturing of micro-algae. [0020] The medium which may be employed in the invention is a known medium which contains natural or artificial seawater as a base, and additives such as a nitrogen source, a phosphorus source, a metal salt, and vitamins. Examples of the nitrogen source include NaNO 3 , KNO 3 , Ca(N0 3

)

2 , NH 4

NO

3 , and (NH 4

)

2

SO

4 . Examples of the phosphorus source include K 2

HPO

4 , KH 2

PO

4 , Na 2

HPO

4 , NaH 2

PO

4 , and sodium glycerophosphate. Examples of the metal salt include NaCl, 15 WO 2012/128396 PCT/JP2012/058505 KCl, CaCl 2 , MgCl 2 , Na 2

SO

4 , K 2

SO

4 , MgSO 4 , Na 2

CO

3 , NaHCO 3 , Na 2 SiO 3 ,

H

3 B0 3 , MnCl 2 , MnSO 4 , FeCl 3 , FeSO 4 , CoCl 2 , ZnSO 4 , CuSO 4 , and Na 2 MoO 4 . Examples of the vitamins include biotin, vitamin B12, thiamine-HCl, nicotinic acid, inositol, folic acid, and thymine. The aforementioned medium may further contain an appropriate additive such as a carbon source or a trace metal, in order to promote production of lauric acid-containing oil or fat. [0021] Examples of preferred media include Daigo IMK medium, f/2 medium, ESM medium, Li medium, and MNK medium. [0022] Preferably, the pH of the thus-prepared medium is adjusted to fall within a range of 7.0 to 8.0 through addition of an appropriate acid or base, and is sterilized in an autoclave before use. [0023] In culturing, no particular limitation is imposed on the amount of algae inoculated to the culture medium. However, the amount is preferably, 1.0 to 10.0% (vol/vol), more preferably 1.0 to 5.0% (vol/vol), with respect to the amount of culturing medium. [0024] No particular limitation is imposed on the culture temperature, so long as the growth of the algae of the present invention is not adversely affected. Generally, the 16 WO 2012/128396 PCT/JP2012/058505 culturing is preferably performed at 10 to 30 0 C, more preferably 15 to 25 0 C. [0025] Light irradiation may be performed under any conditions, so long as photosynthesis can be performed. Needless to say, either artificial light or sunlight may be employed. The illuminance preferably falls within a range of 100 to 50,000 lux, more preferably 300 to 10,000 lux. [0026] The pH during culturing is generally 6.5 to 8.5, preferably 7.0 to 8.0. [0027] Culturing is performed so that an alga is grown in a high density. For example, the culturing period is 7 to 120 days, preferably 7 to 30 days. Any of aeration and agitation culturing, shake culturing, and stationary culturing may be employed. [0028] After completion of culturing, an alga is separated through a customary method such as centrifugation or filtration. The thus-separated alga mass as is, or a broken product thereof obtained through sonication, by means of Dyno Mill or by other means is subjected to solvent extraction with organic solvent such as chloroform, hexane, butanol, methanol, or ethyl acetate, whereby lauric-acid-containing oil or fat can be recovered. [0029] 17 WO 2012/128396 PCT/JP2012/058505 When a Gymnochlora stellata strain CCMP2057 is used, 100 g of the dry alga contain a lauric acid-containing oil or fat in an amount of about 5 to about 10 g. That is, the amount of lauric acid-containing oil or fat produced in 1 L of medium reaches about 0.02 to about 0.05 g. In this case, the oil or fat has a lauric acid content as high as 4.0 to 8.5 weight% of the fatty acid composition. Thus, the amount of produced lauric acid in 1 L of medium is as high as about 0.0008 to about 0.0043 g. [0030] When a Chroomonas diplococca strain strain UTEX LB2422 is used, 100 g of the dry alga contains a lauric acid containing oil or' fat in an amount of about 3 to about 4 g. That is, the amount of lauric acid-containing oil or fat produced in 1 L of medium reaches about 0.007 to about 0.016 g. In this case, the oil or fat has a lauric acid content as high as 5.0 to 17.0 weight% of the fatty acid composition. Thus, the amount of produced lauric acid in 1 L of medium is as high as about 0.0004 to about 0.0027 g. [00311 Separation and recovery of lauric acid from the lauric acid-containing oil or fat may be carried by transforming the oil or fat into a fatty acid mixture or an ester of a fatty acid through a known method; and recovering high concentration of lauric acid through the urea addition method, cooling separation, HPLC, supercritical liquid chromatography, 18 WO 2012/128396 PCT/JP2012/058505 etc. Further, the lauric acid ester can be separated and recovered from the lauric acid-containing oil or fat by esterifying the lauric acid contained in the oil or fat. For example, the lauric acid-containing oil or fat is reacted with alcohol such as methanol under the presence of alkaline catalyst and esters of lauric acid esters can be separated and recovered from the reaction product. Here, examples of lauric acid esters include lower alkyl esters such as methyl ester and ethyl ester, and methyl ester is preferred. Further, lauryl alcohol can be separated and recovered from the lauric acid-containing oil or fat by reducing the lauric acid contained in the oil or fat. For example, the lauric acid-containing oil or fat is hydrogenated under the presence of hydrogenation catalyst and lauryl alcohol can be separated and recovered from the reaction product. [0032] In accordance with the embodiments mentioned above, the present invention discloses the following [1] method for producing lauric acid or esters thereof and [12] method for producing oil or fat containing lauric acid as a constituent fatty acid, and preferably discloses [2]-[11] methods for producing lauric acid or esters thereof and [12]-[14] methods for producing oil or fat containing lauric acid as a constituent fatty acid. 19 WO 2012/128396 PCT/JP2012/058505 [1] Method for producing lauric acid or esters thereof, which method including: culturing at least one species selected from the group consisting of algae in the class Chlorarachniophyceae and algae in the class Cryptophyceae consisting of algae belonging to the genus Rhodomonas and algae belonging to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and Chroomonas placoidea in a medium; recovering, from the culture product, an oil or fat having a lauric acid content of 3 weight% or higher of the fatty acid composition; and, as needed, esterifying the lauric acid in the recovered oil or fat, followed by separating and recovering the lauric acid or esters thereof [2] Method of [1], in which the algae in the class Chlorarachniophyceae is those belonging to the genus Lotharella, Gymnochlora, or Bigelowiella. [3] Method of [1] or [2], in which the algae belonging to the genus Lotharella is Lotharella globosa, Lotharella amoebiformis or Lotharella vacuolata, the algae belonging to the genus Gymnochlora is Gymnochlora stellata and the algae belonging to the genus Bigelowiella include Bigelowiella natans. [4] Method of [3], in which the algae belonging to the genus Lotharella is Lotharella globosa strain CCMP1729, Lotharella amoebiformis strain CCMP2058, Lotharella vacuolata strain CCMP240 or strains having virtually the same phycological properties as those of the 20 WO 2012/128396 PCT/JP2012/058505 algae strains. [5] Method of [3], in which the algae belonging to the genus Gymnochlora is Gymnochlora stellata strain CCMP2057 or strains having virtually the same phycological properties as those of the algae strain. [6] Method of [3], in which the algae belonging to the genus Bigelowiella is Bigelowiella natans strains CCMP621, CCMP2757 or strains having virtually the same phycological properties as those of the algae strains. [7] Method of [1], in which the algae belonging to the genus Chroomonas is Chroomonas diplococca strain UTEX LB2422, Chroomonas mesostigmatica strain NIES1370, Chroomonas nordstedtii strains NIES707, Chroomonas nordstedtii strains NIES NIES710, Chroomonas placoidea strain NIES705, or strains having virtually the same phycological properties as those of the algae strains. [8] Method of [1], in which the algae belonging to the genus Rhodomonas is Rhodomonas salina. [9] Method of [8], in which the Rhodomonas salina is Rhodomonas salina UTEX1375, Rhodomonas salina CCMP272 or strains having virtually the same phycological properties as those of the algae strains. [10] Method of [1]-[9], in which the culturing is performed under light irradiation at an illuminance of 300 to 10,000 for 7-120 days. [11] Method of [1]-[10], in which the lauric acid ester is methyl laurate. 21 WO 2012/128396 PCT/JP2012/058505 [121 Method for producing an oil or fat containing lauric acid as a constituent fatty acid, which method including: culturing, in a medium, at least one species of algae belonging to the genus Chroomonas selected from among Chroomonas diplococca, Chroomonas mesostigmatica, Chroomonas nordstedtii, and Chroomonas placoidea; and recovering, from the culture product, an oil or fat having a lauric acid content of 3 weight% or higher of the fatty acid composition. [13] Method of [12], in which the algae belonging to the genus Chroomonas is Chroomonas diplococca strain UTEX LB2422, Chroomonas mesostigmatica strain NIES1370, Chroomonas nordstedtii strains NIES7O7, Chroomonas nordstedtii strains NIES710, Chroomonas placoidea strain NIES705, or strains having virtually the same phycological properties as those of the algae strains. [14] Method of [12] or [13], in which the culturing is performed under light irradiation at an illuminance of 300 to 10,000 for 7-120 days. [Examples] [0033] Example 1: Culturing of algae in the class Chlorarachniophyceae and analysis of fatty acid composition From "The Provasoli-Guillard National Center for Culture of Marine Phytoplankton (CCMP)," the following 6 algae strains were obtained and employed in the experiments. [0034] 22 WO 2012/128396 PCT/JP2012/058505 [Table 1] Algae strains No. genus/species 1729 Lotharella globosa 2058 Lotharella amoebiformis 240 Lotharella vacuolata 2057 Gymnochlora stellata 621 Bigelowiella natans 2757 Bige/owiella natans [0035] Culturing of algae was performed in the following methods. A commercial medium (Daigo IMK medium, product of Nihon Pharmaceutical Co., Ltd.) (composition, see Table 2) was employed as a seawater medium. [0036] [Table 2] Composition of IMK medium for 1L NaNO 3 200mg Na 2

HPO

4 1.4mg

K

2

HPO

4 5mg

NH

4 Cl 2.68mg Fe-EDTA 5.2mg Mn-EDTA 332pg Na 2 -EDTA 37.2mg ZnSO 4 -7H 2 0 23pg CoSO 4 -7H 2 0 14pg Na 2 MoO 4 -2H 2 0 7. 3pg CuSo 4 -5H 2 0 2 . 5pg

H

2 SeO 3 1.7pg MnCl 2 -4H 2 0 180 pg Thiamin-HC1 200pg Biotin 1.5pg Vitamin B12 1.5pg Artificial sea water 35.96g 23 WO 2012/128396 PCT/JP2012/058505 [0037] Sterilized culture tubes (16 mm x 150 mm) (product of VWR) each plugged with a sponge stopper (60882-167, product of VWR) were used, and a sterilized medium (10 mL/tube) was dispensed to the tubes. Each alga strain (100 pL (in the case of liquid medium) or 1 platinum loop (in the case of solid medium)) was inoculated to a new culture medium. Stationary culturing was performed at room temperature (22 0 C to 24 0 C) under a fluorescent lamp (illuminance: about 3,000 lux, illumination for 12 hours and dark for 12 hours). Through centrifugation of the algaculture at 3,000 rpm for 30 minutes, an alga pellet was obtained. The alga pellet was dried at 80 0 C for about 3 hours to about 16 hours, to thereby obtain dry alga, and the weight of the dry product was measured. The dry product was suspended in 1% saline (0.5 mL), and 5 mg/mL 7-pentadecanone (10 pL) was added as an internal standard to the suspension. Subsequently, chloroform (0.5 mL) and methanol (1 mL) were added to the suspension, and the mixture was vigorously stirred and then allowed to stand for 30 minutes. Thereafter, chloroform (0.5 mL) and 1.5% KCl (0.5 mL) were added to the mixture and stirred, followed by centrifugation at 3,000 rpm for 15 minutes. The formed chloroform layer (lower layer) was recovered by using a Pasteur pipette. [0038] The thus-prepared lipid fraction (about 500 iL) was treated with nitrogen to dryness, and 0.5 N potassium 24 WO 2012/128396 PCT/JP2012/058505 hydroxide/methanol solution (700 pL) was added to the dried fraction, and then incubated at 80 0 C for 30 minutes. Subsequently, 14% boron trifluoride solution (product of SIGMA) (1 mL) was added to the fraction, and then incubated at 80 0 C for 20 minutes. Then, hexane (1 mL) and saturated saline (1 mL) were added to the above mixture, and the mixture was allowed to stand at room temperature for 30 minutes. The thus-obtained hexane layer (upper layer) was recovered and analyzed by GC. (0039] The GC analysis was performed under the following conditions: chromatograph, HP 7890A GC-FID (product of Agilent); column, DB-1 ms 30 m x 200 pm x 0.25 pm (product of J&W scientific); mobile phase, high-purity helium; flow rate, 1 mL/min; and temperature elevation, 100 0 C (1 minute), 5 0 C/min, and 280 0 C (20 minutes) . As saturated fatty acid controls, the following commercial products (all produced from SIGMA) were purchased and analyzed: methyl laurate (C12), methyl myristate (C14), methyl palmitate (C16), and methyl stearate (C18). As unsaturated fatty acid controls, the following commercial products (all produced from SIGMA) were purchased and analyzed: methyl palmitoleate (C16:1), methyl oleate (C18:1), methyl linoleate (C18:2), methyl linolenate (C18:3), methyl eicosapentaenoate (C20:5), and methyl docosahexaenoate (C22:6). Identification of fatty acids was performed on the basis of coincidence in retention time between the fatty acid analyte and the corresponding standard. 25 WO 2012/128396 PCT/JP2012/058505 Lauric acid was also identified by GC-MS. C16 multi unsaturated fatty acids were estimated from the GC-MS analytical results and are represented by C16:x (x is 2 or 3, wherein x represents the number of unsaturated bonds in fatty acid). The GC-MS analysis was performed under the following conditions: chromatograph, HP 7890A GC and 5975C MS (products of Agilent); column, DB-1 ms 30 m x 200 pm x 0.25 ptm (product of J&W scientific); mobile phase, high-purity helium; flow rate, 1 mL/min; and temperature elevation, 100 0 C (1 minute), 5 0 C/min, and 280 0 C (20 minutes) . The amount of a fatty acid ester detected through GC analysis was calculated with reference to the internal standard, and the sum of the amounts of fatty acids was employed as the total fatty acid amount. The value obtained by dividing the total fatty acid amount by the amount of dry alga and multiplying the ratio by 100 was employed as a fatty acid content (%). Table 3 shows the fatty acid compositional data of tested algae species. [0040] [Table 3] Fatty acid composition analysis No. Age Fatty acid composition (%) I Productivity N (days) C12:0 I C14:0 C16:01 C18:01 C18:1] C18:21I8:31 C20:51 022:61 (mg/L) 1729 28 13.0 1.9 25.5 1.8 0.0 4.0 2.0 38.2 13.5 10.0 2058 56 9.3 3.2 40.5 2.0 0.0 1.7 0.0 30.1 13.3 7.0 240 56 3.3 0.0 42.1 2.3 0.0 9.1 0.0 24.7 18.4 4.1 2057 40 8.5 5.1 50.6 6.6 3.6 5.2 0.0 0.0 20.4 35.9 621 33 5.9 8.4 34.1 2.9 7.4 0.0 0.0 32.3 8.9 4.4 2757 33 4.2 8.1 43.5 4.7 9.0 0.0 0.0 22.6 7.9 3.8 26 WO 2012/128396 PCT/JP2012/058505 [0041] Accumulation of lauric acid (>3% of the total fatty acids) was observed in Lotharella globosa strain CCMP1729, Lotharella amoebiformis strain CCMP2058, Lotharella vacuolata strain CCMP240, Gymnochlora stellata strain CCMP2057, and Bigelowiella natans strains CCMP621 and CCMP2757. Particularly, in Gymnochlora stellata strain CCMP2057, high fatty acid productivity and high-level accumulation of lauric acid (about 8.5% of the total fatty acids) were observed. [0042] Example 2: Production of lauric acid using algae in the class Chlorarachniophyceae Gymnochlora stellata CCMP2057 was subjected to stationary culturing in culture tubes (16 mm x 150 mm, containing IMK medium (10 mL)) at room temperature (22 0 C to 24'C) under illumination (illuminance: about 3,000 lux, illumination for 12 hours and dark for 12 hours) for four weeks, to thereby produce a seed culture liquid. The seed culture liquid was inoculated into IMK medium (100 mL) at 2% (v/v) placed in a 200-mL Erlenmeyer flask, and stationary culturing was performed at room temperature (22 0 C to 24 0 C) under illumination (illuminance: about 3,000 lux, illumination for 12 hours and dark for 12 hours) for 31 days. The culture liquid was centrifuged at 3,000 rpm for 30 minutes, to thereby recover cells, which were then washed once with 1% (w/v) aqueous sodium chloride solution. [0043] 27 WO 2012/128396 PCT/JP2012/058505 The alga which had been recovered from the culture liquid (100 mL) was dried at 80 0 C for about 16 hours, and chloroform (2 mL) and methanol (4 mL) were added to the dried alga. The mixture was vigorously stirred and then allowed to stand for 30 minutes. Thereafter, chloroform (2 mL) and 1.5% KCl (2 mL) were added thereto, and the obtained mixture was stirred. The stirred mixture was centrifuged at 3,000 rpm for 15 minutes, and the chloroform layer (lower layer) was collected by using a Pasteur pipette. An aliquot (100 pL) was recovered from the collected chloroform layer and dried to solid through nitrogen gas sprayed thereto. The dried product was dissolved in chloroform (10 pL) . An aliquot (1 pL) was sampled from the chloroform solution, and the neutral fat content thereof was determined by means of Iatroscan (product of Mitsubishi Kagaku Iatron, Inc.). As a result, neutral lipid (1.2 mg) was obtained from the culture liquid (100 mL). [0044] Through a methyl esterification method similar to that described in Example 1, an aliquot (500 pL) of the above collected chloroform layer was analyzed. As a result, the total amount of the fatty acids obtained from the culture liquid (100 mL) was 6.2 mg, and the lauric acid content of the total fatty acid was 4.9%. That is, lauric acid (0.3 mg) was recovered from the culture liquid (100 mL). [00451 Example 3: Culturing of algae belonging to the genus 28 WO 2012/128396 PCT/JP2012/058505 Chroomonas and analysis of fatty acid composition From the Culture Collection of Algae at University of Texas at Austin (UTEX) and National Institute for Environmental Studies (NIES), the following 9 algae strains belonging to the genus Chroomonas were obtained and employed in the experiments. [0046] [Table 4] Algae strains Organization No. genus/species UTEX LB 2422 Chroomonas dip/ococca NIES 1004 Chroomonas coerulea NIES 714 Chroomonas coeru/ea NIES 704 Chroomonas dispersa NIES 1370 Chroomonas mesostigmatica NIES 707 Chroomonas nordstedtil NIES 710 Chroomonas nordstedtii NIES 705 Chroomonas p/acoidea NIES 2331 Chroomonas sp. [0047] Culturing of algae was performed in the following methods. C medium (composition, see Table 5) and WA medium (composition, see Table 6) were employed as fresh water media, and f/2 medium (composition, see Table 7) and a commercial medium (Daigo IMK medium, product of Nihon Pharmaceutical Co., Ltd.) (composition, see Table 8) were employed as seawater media. [0048] 29 WO 2012/128396 PCT/JP2012/058505 [Table 51 Composition of C medium for IL Ca(N0 3

)

2 -4H 2 0 150 mg

KNO

3 100 mg P-Na 2 glycerophosphate-5H 2 0 50 mg MgSO 4 -7H 2 0 40 mg Vitamin B12 0.1 pig Biotin 0.1 g Thiamine HCI 10 pig *PIV metal mixture solution 3 mL Tris(hydroxymethyl)aminomethane 500 mg pH 7.5 *PIV metal mixture solution FeCl3-6H 2 0 19.6 mg MnC12-4H 2 0 3.6 mg ZnSO 4 -7H 2 0 2.2 mg CoCl 2 -6H 2 0 0.4 mg Na 2 MoO 4 -2H 2 0 0.25 mg Na 2 EDTA -2H 2 0 100 mg Distilled water 100 mL [0049] 30 WO 2012/128396 PCT/JP2012/058505 [Table 6] Composition of WA medium for 1L NaNO 3 20mg Ca(N0 3

)

2 -4H 2 0 60mg KCI 10mg MgSO 4 -7H 2 0 20mg Na 2 glyceroPO 4 10mg Na 2 EDTA 5mg FeCl3-6H 2 O 240pg

H

3

BO

3 1mg MnCl2-4H 2 0 7.2mg ZnCl 2 50pg CoCl2-6H 2 0 20pg Tris amino 100mg Thiamin-HCI 10pg Biotin 10pg Vitamin B12 1Opg [0050] [Table 7] Composition of f/2 medium for IL NaNO 3 75 mg NaH 2

PO

4 -2H 2 0 5 mg Vitamin B12 135 ptg Biotin 25 pg Thiamine HCI 1.1 mg Na 2 SiO 3 -9H 2 0 30 mg *f/2 metal mixture solution 1 mL Artificial sea water 999 mL *f/2 metal mixture solution Na 2 EDTA -2H 2 0 880 mg Fe(NH 4

)

2

(SO

4

)

2 -6H 2 0 920 mg CoSO 4 -7H 2 0 2.8 mg ZnSO 4 -7H 2 0 4.6 mg MnSO 4

-H

2 0 30.6 mg CuC12 -2H20 1.36 mg Na 2 MoO 4 -2H 2 0 1.46 mg Distilled water 100 mL 31 WO 2012/128396 PCT/JP2012/058505 [0 051] [Table 8] Composition of IMK medium for 1L NaN03 200mg Na2HPO4 1.4mg K2HPO4 5mg NH4CI 2.68mg Fe-EDTA 5.2mg Mn-EDTA 332pg Na2-EDTA 37.2mg ZnSO4-7H20 23pg CoSO4-7H20 14pg Na2MoO4-2H20 7.3Pg CuSO 4 -5H 2 0 2.5pg

H

2 SeO 3 1.7pg MnC12-4H 2 0 180pg Thiamin -HCI 200pg Biotin 1.5pg Vitamin B12 1.5pg Artificial sea water 35.96g [0052] Sterilized culture tubes (16 mm x 150 mm) (product of VWR) each plugged with a sponge stopper (60882-167, product of VWR) were used, and a sterilized medium (10 mL/tube) was dispensed to the tubes. Each alga strain (100 ptL (in the case of liquid medium) or 1 platinum loop (in the case of solid medium)) was inoculated to a new culture medium. Stationary culturing was performed at room temperature (22 0 C to 24'C) under a fluorescent lamp (illuminance: about 3,000 lux, illumination for 12 hours and dark for 12 hours). Through centrifugation of the alga culture at 3,000 rpm for 30 minutes, an alga pellet was obtained. The alga pellet 32 WO 2012/128396 PCT/JP2012/058505 was dried at 80'C for about 3 hours to about 16 hours, to thereby obtain dry alga, and the weight of the dry product was measured. The dry product was suspended in 1% saline (0.5 mL), and 5 mg/mL 7-pentadecanone (10 pL) was added as an internal standard to the suspension. Subsequently, chloroform (0.5 mL) and methanol (1 mL) were added to the suspension, and the mixture was vigorously stirred and then allowed to stand for 30 minutes. Thereafter, chloroform (0.5 mL) and 1.5% KCl (0.5 mL) were added to the mixture and stirred, followed by centrifugation at 3,000 rpm for 15 minutes. The formed chloroform layer (lower layer) was recovered by using a Pasteur pipette. [0053] The thus-prepared lipid fraction (about 500 pL) was treated with nitrogen to dryness, and 0.5 N potassium hydroxide/methanol solution (700 piL) was added to the dried fraction, and then incubated at 80 0 C for 30 minutes. Subsequently, 14% boron trifluoride solution (product of SIGMA) (1 mL) was added to the fraction, and then incubated at 80 0 C for 20 minutes. Then, hexane (1 mL) and saturated saline (1 mL) were added to the above mixture, and the mixture was allowed to stand at room temperature for 30 minutes. The thus-obtained hexane layer (upper layer) was recovered and analyzed by GC. [00541 The GC analysis was performed under the following conditions: chromatograph, HP 7890A GC-FID (product of 33 WO 2012/128396 PCT/JP2012/058505 Agilent); column, DB-1 ms 30 m x 200 pLm x 0.25 ptm (product of J&W scientific); mobile phase, high-purity helium; flow rate, 1 mL/min; and temperature elevation, 100 0 C (1 minute), 5 0 C/min, and 280 0 C (20 minutes) . As saturated fatty acid controls, the following commercial products (all produced from SIGMA) were purchased and analyzed: methyl laurate (C12), methyl myristate (C14), methyl palmitate (C16), and methyl stearate (C18). As unsaturated fatty acid controls, the following commercial products (all produced from SIGMA) were purchased and analyzed: methyl palmitoleate (C16:1), methyl oleate (C18:1), methyl linoleate (C18:2), methyl linolenate (C18:3), methyl eicosapentaenoate (C20:5), and methyl docosahexaenoate (C22:6). Identification of fatty acids was performed on the basis of coincidence in retention time between the fatty acid analyte and the corresponding standard. Lauric acid was also identified by GC-MS. C16 multi unsaturated fatty acids were estimated from the GC-MS analytical results and are represented by C16:x (x is 2 or 3, wherein x represents the number of unsaturated bonds in fatty acid). The GC-MS analysis was performed under the following conditions: chromatograph, HP 7890A GC and 5975C MS (products of Agilent); column, DB-1 ms 30 m x 200 pim x 0.25 ptm (product of J&W scientific); mobile phase, high-purity helium; flow rate, 1 mL/min; and temperature elevation, 100 0 C (1 minute), 5 0 C/min, and 280 0 C (20 minutes). The amount of a fatty acid ester detected through GC analysis was calculated with reference to the internal standard, and the sum of the 34 WO 2012/128396 PCT/JP2012/058505 amounts of fatty acids was employed as the total fatty acid amount (g). The fatty acid productivity (g/L or mg/L) was obtained by dividing the total fatty acid amount by the volume of culture liquid (L) . The value obtained by dividing the amount of each fatty acid by the total amount of the fatty acids and multiplying the ratio by 100 was employed as a fatty acid content (%). Table 9 shows the fatty acid compositional data of tested algae species. [0055] [Table 9] Fatty acid composition analysis No. Medium date Fatty acid composition Productivity No.I - I dat C : C14:( C16:( C16:1] 16:31 C18:01 C I 8: C1 C8:2[ C18:3C20:5 C226 (m /L) LB 2422 IMK 30 16.9 24.7 7.4 2.3 0.0 1.5 6.5 6.2 14.5 14.0 6.2 15.66 1004 C 47 0.0 0.0 29.8 0.0 0.0 11.3 0.0 10.9 33.9 14.2 0.0 5.9 714 WA 47 0.0 4.6 24.4 0.0 0.0 19.9 0.0 24.5 14.7 7.3 4.7 8.2 714 C 47 0.0 4.8 19.9 4.0 0.0 9.8 9.8 30.7 0.0 13.4 7.7 6.0 704 IMK 47 0.0 0.0 35.7 0.0 0.0 9.2 7.9 5.6 20.3 13.7 7.6 8.9 1370 IMK 47 9.2 8.9 21.5 0.0 0.0 10.3 15.0 18.9 6.6 9.7 0.0 7.7 f/2 47 11.1 9.6 27.7 0.0 0.0 8.8 0.0 20.4 3.0 14.1 5.3 10.8 707 C 47 6.3 10.0 20.7 9.0 6.4 10.3 60 18.3 6.0 7.1 0.0 7.0 710 C 47 6.9 0.0 21.8 0.0 0.0 5.2 0.0 30.6 0.0 25.6 10.0 7.3 705 IMK 47 7.7 14.3 24.9 0.0 0.0 4.8 3.3 13.2 10.1 16.3 5.5 11.7 2331 f/2 47 4.4 6.1 27.5 0.0 0.0 7.2 8.8 6.6 26.9 12.3 0.0 11.5 [0056] Accumulation of lauric acid (>3% of the total fatty acids) was observed in Chroomonas diplococca strain UTEX LB2422, Chroomonas mesostigmatica strain NIES1370, Chroomonas nordstedtii strain NIES707, Chroomonas nordstedtii strain NIES710, and Chroomonas placoidea strain NIES705. Particularly, in Chroomonas diplococca strain UTEX LB2422, a 35 WO 2012/128396 PCT/JP2012/058505 very high-level accumulation of lauric acid (about 17% of the total fatty acids) was observed. [0057] Example 4: Production of lauric acid using algae belonging to the genus Chromonas An oil or fat having high lauric acid content was produced in the following manner. Chroomonas diplococca (strain LB2422) was subjected to stationary culturing in culture tubes (16 mm x 150 mm, containing IMK medium (10 mL)) at room temperature (22 0 C to 24'C) under illumination (illuminance: about 3,000 lux, illumination for 12 hours and dark for 12 hours) for four weeks, to thereby produce a seed culture liquid. The seed culture liquid was inoculated into IMK medium (100 mL) at 2% (v/v) placed in a 200-mL Erlenmeyer flask, and stationary culturing was performed at room temperature (22 0 C to 24 0 C) under illumination (illuminance: about 3,000 lux, illumination for 12 hours and dark for 12 hours) for 31 days. The culture liquid was centrifuged at 3,000 rpm for 30 minutes, to thereby recover cells, which were then washed once with 1% (w/v) aqueous sodium chloride solution. [0058] The alga which had been recovered from the culture liquid (100 mL) was dried at 80'C for about 16 hours, and chloroform (2 mL) and methanol (4 mL) were added to the dried alga. The mixture was vigorously stirred and then allowed to stand for 30 minutes. Thereafter, chloroform (2 mL) and 1.5% 36 WO 2012/128396 PCT/JP2012/058505 KCl (2 mL) were added thereto, and the obtained mixture was stirred. The stirred mixture was centrifuged at 3,000 rpm for 15 minutes, and the chloroform layer (lower layer) was collected by using Pasteur pipette. An aliquot (100 ptL) was recovered from the collected chloroform layer and dried to solid through nitrogen gas sprayed thereto. The dried product was dissolved in chloroform (10 pL) . An aliquot (1 pL) was sampled from the chloroform solution, and the neutral fat content thereof was determined by means of Iatroscan (product of Mitsubishi Kagaku Iatron, Inc.) . As a result, neutral lipid (0.64 mg) was obtained from the culture liquid (100 mL). [0059] Through a methyl esterification method similar to that described in Example 3, an aliquot (500 pL) of the above collected chloroform layer was analyzed. As a result, the total amount of the fatty acids obtained from the culture liquid (100 mL) was 3.5 mg, and the lauric acid content of the fatty acids was 5.7%. That is, lauric acid (0.2 mg) was recovered from the culture liquid (100 mL). Example 5: Culturing of algae belonging to the genus Rhodomonas and analysis of fatty acid composition As algae belonging to the genus Rhodomonas, Rhodomonas salina UTEX1375 and Rhodomonas salina CCMP272 were purchased from The culture collection of algae at University of Texas at Austin (UTEX) and The Provasoli-Guillard National Center 37 WO 2012/128396 PCT/JP2012/058505 for Culture of Marine Phytoplankton (CCMP), and these algae strains were tested by using an IMK medium through the method similar to that employed in Example 3. The test procedure of Example 3 was repeated, except that the culture times described in Table 9 were employed. The total fatty acid productivity and the ratio of each fatty acid were determined. The test has revealed that both algae strains had lauric acid contents of 9.4% and 8.8% respectively, which are higher than 3%. [00611 [Table 10] Fatty acid composition analysis No. genus/species medium Week: Fat cdc osto roductivity UTEX 1375 Roooa au IK 7.5 9. 14 52. 12 8 00 R . . 590 CCMP 272 4 8onas sauna MK 8.8 9.4 24.7 106 10.0 0.0 9.3 14.6 330 38

Claims (1)

1. 8.0 and is sterilized in an autoclave before the culturing. [Claim 13] 15 An oil or fat comprising lauric acid as a constituent fatty acid, produced by the method of any one of claims 9 to 12. 44