BR122014033084B1 - METHOD FOR ANALYZING PROPERTIES OF SOYBEAN ODOR - Google Patents

Description

Invention Patent Descriptive Report for "METHOD FOR ANALYZING SOY ODOR PRODUCING PROPERTIES".

Divided from PI0513067-0, filed July 7, 2005. Background of the Invention This patent application claims the priority of provisional patent application Serial No. 60 / 521,846, filed July 9, 2004, the entire contents of which are specifically incorporated herein by reference.

Field of the Invention The present invention relates to the field of nutrition and food sciences. Particularly, the invention relates to soybean compositions having better organoleptic properties, such as lower odor, and methods for their use and production.

Description of Prior Art Soya beans provide high quality proteins that provide health benefits to humans (Hermansen et al, 2003; Bazzano et al, 2001; Food and Drug Administration, 1999). Demand for soybeans to make soy foods has not risen as much as expected in the last three decades (Wolfe and Cowan, 1975; and "Soysource", The United Soybean Board, 1999). This is partly due to the undesirable odor associated with soy products (McLeod and Ames, 1988; and Freese, 1999). The undesirable odor of soybeans is commonly described as "grainy". The components that confer the grain characteristic of soybeans include many volatile fatty acids, aliphatic carbonyls, amines, alcohols, aldehydes and holes in the enzymatic action derivatives on various compounds found in soybeans and their later oxidation which is caused by many mechanisms (Wolfe and Cowan, 1975; Sesssa and Rackis, 1977).

[005] Kobayashi et al. (1995) concluded that the major contributors to the odor of unboiled soymilk were (trans, trans) -2,4-nonadienal, (trans, trans) -2,4-decadienal, hexanal, 2-pentyl furan , 1-octen-3-one, (trans) -2-nonenal, and (trans, cis) -2,4-nonadienal. The most intense odors extracted from heat-treated soy milk were identified as (trans, trans) -2,4-decadienal and n-hexanal (Feng, Cornell University Ph.D. Dissertation, 2000). (Trans, trans) -2,4-decadienal formation occurs at a low rate at room temperature (Frankel, 1988); however, this reaction is intensified because of thermal degradation during soybean processing under hot conditions (Lin, 2003). Other contributors to odors were (trans) -4,5-epoxy (E) -decenal (formed from 2,4-decadienal), (trans, cis) -2,6-nonadienal, (trans) -2- nonenal, (trans, trans) -2,4-nonadienal, 2,4-nonadienal, maltol, vanillin and β-damascenone. The most potent odorants in soymilk, determined by the volume of minimum void space needed to detect by olfactometry, were hexanal, acetaldehyde, methanethiol, dimethyl trisulfide, and 2-methyl furan (Boatright, 2002).

The most intense odorants in soy protein isolates have been identified as dimethyl trisulfide, (trans, trans) -2,4-decadienal, 2-pentylpyridine, (trans, trans) -2,4-nonadienal, hexanal, acetophenone , and 1-octen-3-one (Boatright and Lei, 1999). The mechanism of methanethiol and dimethyl trisulfide formation involves free radicals formed by lipid oxidation (Lei and Boatright, 2003), and enzyme products such as cysteine synthase (Boatright, 2003, poster 45C-26, IFT annual meeting , Chicago).

The formation of 2-pentyl pyridine occurs from a spontaneous reaction between 2,4-decadienal and ammonia at room temperature. The free amino acids arginine, lysine, asparagine and glutaminase increase the formation of 2-pentylpyridine probably by providing ammonia during soy protein processing (Zhou and Boatright, 2000; Kim et al., 1996). Therefore, free amino acids may also form other undesirable products. Exposure of asparagine and glucose to high temperatures results in the formation of acrylamide (Jung et al., 2003). Asparagine exposed to cooking temperatures can form mutagens (Knize et al., 1994). Free arginine was enriched in soybeans lacking β-conglycinins and glycinins (Takahashi et al., 2003).

Once formed, odors are difficult to remove from soy ingredients because they are associated with protein (Franzen and Kinsella, 1974). The quality of natural flavors added to soy foods is also unfavorably changed because some of the odors bind to soy protein. The carbonyl and 2-pentylpyridine compounds bound with greater affinity to glycine fractions than β-conglycinin fractions (Zhou et al., 2002; O'Keefe et al., 1991). Extraction of oil-body associated proteins and polar lipids significantly reduced the amount of odor associated with soy protein isolates (Samoto et al., 1998).

[009] Textures created by protein-protein interactions may have more effect on intensity than nose odor concentration (Weel et al., 2002). Soy proteins may contribute to the low organoleptic quality of soy beverages by forming insoluble aggregates and sandy mouthfeel (Skarra and Miller, 2002). Among the main soy proteins, glycinins are more sensitive to pH and Ca + 2 -induced insolubilization (Yuan, 2002) and soybeans that contain a low glycine to β-conglycinin ratio are useful for creating soy protein ingredients. soluble (US Patent No. 6,171,640).

Lipid oxidation reactions also influence protein solubility. Antioxidants may be added during the manufacture of soy protein isolates to limit free radical-induced protein oxidation and improve soluble protein yield (US Patent 5,777,080). Some peptides may react during polysaccharide processing to form antioxidant compounds (Matsumara, 2003).

Color influences perceptions of freshness and flavor (Joshi, 2000). Low amounts of reducing sugar and aldehydes formed from lipid oxidation react with heating amino groups of proteins to form brown pigments by Maillard's tanning reaction (Kwok et al., 1999). Soymilk with a higher aldehyde content will create a darker, less attractive color after thermal processing. On the other hand, lipid oxidation during soymilk processing discolour yellow pigments in soymilk (Obata and Matsuura, 1997).

Soybeans are refined to improve flavor by extracting lipids and other components by alcohol extraction, enzyme treatments, protein clot washing, protein ultrafiltration and / or instant spraying. These processes increase the cost of soy protein ingredients, and typically lower the amounts of healthy components that become available (such as fiber, oligosaccharides, isoflavones, polyunsaturated fatty acids, tocopherols, phospholipids, bioactive peptides). The processing approaches used to improve the organoleptic properties of soy protein ingredients are limited in effectiveness by soy protein-linked odors and odor-promoting conditions (pH 8-10). Deprived soybeans from one to three of lipoxygenases 1, 2 and 3 were created using mutation generation to reduce granular odor formation (Hajika et al, 1991). The aroma analysis of soymilk and soy flour made from soybeans lacking the three lipoxygenases was found to contain lower amounts of various odors, but higher amounts of 1-octen-3-ol than the lineage. originating soybean containing the three lipoxygenases (Hao et al, 2002). Similar levels of 2,4-decadienal were found in defatted flour and soy protein isolate, made from one soybean seed lacking the three lipoxygenases and two other soybean strains (Boatright et al., 1998). Soybean foods prepared from lipoxygenase-deficient soybeans tasted better compared to soybean-based foods from control soybeans (Wilson, 1996). Soymilk prepared from lacking soybeans from the three lipoxygenases was perceived as bitter than control, especially after 15 months of seed storage, but this difference was expected to be eliminated by adding sugar (Torres-Penaranda and Reitmeier, 2001).

Transgenic modifications are proposed to improve the taste of soybeans by reducing the levels of polyunsaturated fatty acids (U.S. Patent 5,981,781), lipoxygenases (U.S. Patent Application 2003/0074693 and / or hydroperoxide lyases (U.S. Pat. No. 6,444,874) Soybeans containing less than 10% polyunsaturated fatty acids and more than 75% oleic fatty acids produce a frying oil that is less palatable than oil with more fatty acids. polyunsaturated (Warner et al, 2001).

Chemicals such as polyphosphates (US Patent No. 6,355,296) may be used to limit the production of aroma deviations and improve protein solubility. Other additives, such as gallic acid (PCT No. WO 01/06866) or aldehyde oxidase (Maheshwari et al., 1997) may be used to remove odors.

There is little published information on the effect of natural genetic variations on the flavor and color attributes of soybeans. The thiobarbituric acidity index for 16 soybean varieties was determined as a measure of lipid oxidation and no correlation was found with the soybean vitamin E content (Dahuja and Madaan, 2004). The amounts of 2-pentyl pyridine and 2,4-decadienal in soy flour and soy protein isolate made from three soybean varieties were determined (Zhou and Boatright, 1999). The effects of drying conditions on the removal of green pigment, chlorophyll, from soybeans were studied (Salete etal., 2003; Sinnecker etal., 2002).

In recent decades scientists have shown that oils prepared from lipoxygenase-deficient soybeans had no better oxidation stability. Soybean proteins produced from lipoxygenase-free soybeans still contained significant levels of granular flavor (Maheshwari et al., 1997).

[0017] The first step in making soy milk or soy protein ingredients is to peel (or decorticate) soybeans to create soybean pulps. Hypocotyls can also be separated from cotyledons. Soybean pulps are defined as peeled soybeans and may or may not include cotyledons. A method for preparing the pulps is described, for example, in US Patent 5,727,689. A method of peeling includes, but is not limited to, draining the seeds between countercurrent rollers or a fractionating mill and vacuuming light peels leaving the pulps. The pulps may be soaked in water to produce soy milk or flocked and extracted using hexane as an initial step in the manufacture of defatted soy flour, soy protein concentrates, soy protein isolates and purified protein fractions as desired.

The present invention provides a novel method for determining the ability of soybean pulp to resist the production of important odorant compounds identified as 2,4-decadienal, hexanal, hexanol and 1-octen-3-ol. These compounds were selected as indicators of the extent of different types of oxidation reactions. Hexanal and hexanol result from the decomposition of compounds containing hydroperoxides (peroxides at positions 9 and 12 of the fatty acid carbons) by hydroperoxide lyases and alcohol dehydrogenases. 2,4-decadienal is a degradation product of the lipoxygenase pathway, which admittedly does not involve hydroperoxide lyases. 1-Octen-3-ol is formed by the action of hydroperoxide lyases on hydroperoxides formed at the 10 position of the linoleic acid carbon. These compounds may react further by further processing to form more potent odors. For example, 2,4-decadienal is involved in the formation of 2-pentyl pyridine and 1-octen-3-ol is involved in the formation of 1-octen-3-one. Summary of the Invention In one aspect, the invention provides a soy meat composition made from soy beans comprising lipoxygenases 1, 2 and 3, wherein the composition comprises more than 10% linoleic acid as a percentage of total fatty acids and less than 20 xg of hexane plus 2,4-decadienal plus total hexanol per gram after oxidation under mild aqueous conditions. The composition may or may not comprise lipoxygenases, or any combination thereof, and may comprise deactivated lipoxygenase. In one embodiment, the composition comprises lipoxygenase-2. In certain embodiments, the composition provided by the invention may comprise less than about 15 µg or less than about 18 µg of more hexanal plus 2,4-decadienal plus total hexanol. In other embodiments, the composition may comprise about 6g to about 20g, about 10g to about 20g, or about 12 to 18g of more hexanal 2,4-decadienal plus total hexanol. In other embodiments, the composition may comprise less than 4% linolenic acid as a percentage of total fatty acids, including less than about 3% and between about 1% and 4% or about 2% to about 4%. .

In another embodiment, a composition of the invention may comprise less than 2,000 µg per gram of free arginine and / or less than 400 µg free asparagine per gram, including less than about 1,800 µg per gram. free arginine and / or less than about 350 µg free asparagine per gram. Such a composition, in certain embodiments, may comprise from about 300 to about 2,000 grams per gram of free arginine, including about 500-2,000, about 1,200-1,800, and about 1,000-2,000 grams per gram of arginine. free. A composition provided by the invention may further, in certain embodiments, comprise from about 50 µg to about 400 µg of free asparagine per gram, including about 100-400, 100-350, 200-400, 300-400, and 250 µg. -400 cg free asparagine per gram.

In still another embodiment, the compositions provided by the invention may have a color measured as b * value less than 30 and an L * value greater than 80, monitored by the CIE-L * a * b * system, where L * indicates luminosity and b * indicates the hue on a blue (-) to yellow {+) axis. In certain embodiments, a composition of the invention may comprise a color measured as b * value of about 18-30, about 20-30, about 25-30, and less than about 25. In other embodiments, a The composition provided by the invention may comprise an L * value of about 80-100, about 80-90 and greater than about 90. In certain embodiments, a composition provided by the invention may comprise less than 8 µg of 1-1 µm. octen-3-ol per gram after oxidation under mild aqueous conditions, including less than about 6g, less than about 5g, between about 1.3 and about 8g, between about 2 and about 8g. 8g, and between about 4 and about 8g. A composition provided by the invention may also have more than 30% protein as β-conglycinin and may have less than 25% protein as glycine. Such a composition may further be defined as having more than about 40% of the protein as β-conglycinin, and having a β-conglycinin content between about 30% and about 60%, about 40% to 60%, about from 35% to 55%, and about 30% to about 50% of the protein. Such a composition may further be defined as having a glycine content of less than about 20%, 15% and 10%, and may comprise from about 0% to 25%, 5% to 20%, 1% to 25%, and about 10-25% of the protein as glycine.

In another aspect of the invention there is provided a soybean composition having more than 30% protein as β-conglycinin and less than 25% protein as glycine, less than 5,000 µg per gram of protein. free arginine, and less than 900 µg per gram of asparagine. Such a composition may comprise, in certain embodiments, from about 300 to 5,000 cg per gram, between about 1,000 and 5,000 cg per gram, between about 3,000 and 5,000 cg per gram, between about 1,000 and 4,000 cg per gram, and between about 500 and 2,000 ocg per gram of free arginine. Such a composition may comprise, in certain embodiments, less than 400 µg per gram of free asparagine, between about 50 and 400 µg per gram, between about 100 and 400 µg per gram, between about 100 and 700 µg per gram, and between about 200 and 900 µg per gram of free asparagine. In one embodiment, the composition is less than 2,000 µg per gram of free arginine and less than 400 µg per gram of free asparagine.

In another embodiment, the composition provided comprises less than 4 µg per gram of 1-octen-3-ol after oxidation under mild aqueous conditions, including less than about 3 µg, from about 1.3 µg. and 3 xg, between about 1.3 and 4g, and between about 2g and 4g per gram. In still other embodiments, the composition has a concentration of linolenic acid of between 1% and 14% of total fatty acids, including about 3 to 14%, about 5 to 14%, about 1.5% to 12%, about from 3% to 12%, and about 7 to 14%. In yet another embodiment, the composition has a concentration of 10% to 60% of the total fatty acids, including about 10% to 50%, about 10% to 40%, about 15% to 60%. %, between about 20% and 50%, and between about 20% and 60%.

In certain embodiments, a soy meat composition provided by the invention may be defined as lacking one or more lipoxygenases. In one embodiment, a soybean meat composition provided by the invention may be defined as lacking lipoxygenase-2. In other embodiments, any combination of lipoxygenase-1, lipoxygenase-2 and / or lipoxygenase-3 is absent, including any two or all three of these lipoxygenases. A composition of the invention may also be defined as having a color characterized by a b * value less than 30 and an L * value greater than 80, monitored by the C1E-L * a * b * system, where L * indicates luminosity and b * indicates the hue on a blue (-} to yellow (+) axis. In still other embodiments, a composition provided by the invention may comprise 67-69 mg lysine per gram protein, may comprise 72-80 mg arginine per gram of protein and / or may comprise 28-30 mg histidine per gram of protein.

In yet another aspect, the invention provides a method for analyzing the odor producing properties of a soybean, comprising determining the level of at least one compound selected from the group consisting of 2,4-decadienal, hexanal hexanol, and 1-octen-3-ol. In one embodiment, the method may comprise determining the level of the compound, which comprises incubating a mixture of about 1 part soy flour and about 4 parts water for a period in the range of about 1 to about 40 minutes. and quantifying the amounts of at least one compound selected from the group consisting of 2,4-decadienal, hexanal, hexanol, and 1-octen-3-ol, and combinations thereof, using deuterated standards for hexanal, hexanol and decadienal. Soy flour can be made from peeled soy beans.

In yet another aspect, the invention provides a method for obtaining a soybean variety that produces soybeans and soybean meats with diminished odor producing properties, comprising measuring the level of at least one compound selected from the group that contains the same. consists of 2,4-decadienal, hexanol, hexanal, 1-octen-3-ol, and any combination thereof, in one or more soya beans or soya meats of the first and second soya bean varieties, and select the variety which produces seeds with lower levels of compost. The method may further comprise crossing a plant of the selected variety with a different plant to produce progeny and measuring the level of at least one compound selected from the group consisting of 2,4-decadienal, hexanol, hexanal, 1-octen-3. -ol, and any combination thereof, in one or more soya beans or progeny soya beans.

In yet another aspect, the invention provides a method for selecting a soybean variety that resists fungal contamination, comprising selecting a variety comprising less than 5 µg of 1-octen-3-ol per gram, measured incubating a mixture of about 1 part soy flour and about 4 parts water over a period of about 1 to about 40 minutes, and measuring 1-octen-3-ol.

In yet another aspect, the invention provides a seed of soybean plant designated 0119149, whose representative seed has been deposited under ATCC accession number PTA-6197. The invention further provides a soybean plant 011949 or parts thereof produced by cultivating such seed. Such a plant of the invention may comprise a transgene. In still other embodiments, the invention provides a method for producing a soybean plant derived from the 0119149 soybean plant, which method comprises the steps of: (a) preparing a progeny plant derived from the 0119149 soybean plant by crossing a soybean plant soybean 0119149 with a second soybean plant, where a seed sample of the soybean plant 0119149 was deposited under ATCC accession number PTA-6197; (b) crossbreeding the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation; (c) developing a progeny plant of a subsequent generation from said seed and crossing the progeny plant of a subsequent generation with itself or a second plant; and (d) repeating steps (b) and (c) for a 3-10 generation addition to produce an inbred soybean plant derived from soybean plant 0119149.

In yet another aspect, the invention provides soybeans with better organoleptic properties (i.e. soybeans with better taste, color, odor and mouthfeel properties) after oxidation under mild aqueous conditions. Soybeans with low amounts of free arginine and asparagine are also provided to improve the organoleptic properties of soybeans. In another embodiment, soybeans with reduced levels of linoleic and linolenic acids are provided to improve organoleptic properties.

A soybean plant provided by the invention may comprise, in one embodiment, one or more transgenes. Examples include a herbicide resistance gene that will produce herbicide resistant plants and an insect resistance gene.

According to the invention, soybean seeds containing lipoxygenases 1, 2 and 3, and more than about 10% linoleic acid as a percentage of total fatty acids, which produce less than 20 µg are provided. of 2,4-decadienal plus hexanal plus total hexanol per gram of ground seeds after oxidation under mild aqueous conditions.

According to another aspect of the invention, lipoxygenase-containing soybeans are provided, having less than about 4% linolenic fatty acid and more than about 10% linoleic acid as a percentage of total fatty acids, and which produce less than 20g of 2,4-decadienal plus hexanal plus total hexanol per gram of ground seeds after oxidation under mild aqueous conditions. The same soybeans may also produce less than 8 µg of 1-octen-3-ol per gram of ground seeds after oxidation under mild aqueous conditions.

According to yet another aspect of the invention, the invention provides soybeans having less than about 2,000 cg free arginine and less than about 400 cg free asparagine per gram of dry seed, and which produce less than about 20 µg / g of hexane plus 2,4-decadienal plus total hexanol per gram of ground seeds after oxidation under mild aqueous conditions. The same seeds may also produce less than 8 µg of 1-octen-3-ol per gram of ground seeds after oxidation under mild aqueous conditions.

According to yet another aspect of the invention, soybeans are provided which have a yellow color, measured as a "b * value", of less than 30, and which produce less than 20 µg of 2,4- decadienal plus hexanal plus total hexanol per gram of ground seeds after oxidation under mild aqueous conditions. The same seeds may also produce less than 8 µg of 1-octen-3-ol per gram of ground seeds after oxidation under mild aqueous conditions.

According to yet another aspect of the present invention, there are provided soybeans having more than 30% protein as β-conglycinin and / or less than 25% protein as glycinins, which produce less than 20 µg / g 2,4-decadienal plus hexanal plus total hexanol per gram of ground seeds after oxidation under mild aqueous conditions. The same seeds may also produce less than 8 µg of 1-octen-3-ol per gram of ground seeds after oxidation under mild aqueous conditions.

According to the present invention, soybeans comprising less than 5,000 µg free arginine, less than 900 µg free asparagine, and more than more than 30% protein such as β-conglycinin and less than 25% protein as glycines, which produce less than 20 µg / g of hexane plus 2,4-decadienal plus total hexanol per gram of ground seeds after oxidation under mild aqueous conditions. The same seeds may also produce less than 8 µg of 1-octen-3-ol per gram of ground seeds after oxidation under mild aqueous conditions.

According to yet another aspect of the invention, soybeans resulting from a crossover of a first soybean seed having more than more than 30% protein as β-conglycinin and less than 25% soybean are provided. protein such as glycine, and a second soybean seed that produces less than 20 µg / g of hexane plus 2,4-decadienal plus hexanol per gram of ground seeds after oxidation under mild aqueous conditions.

According to yet another aspect of the invention, soybeans are provided as a result of crossing a first soybean seed which has less than 4% linolenic fatty acid and more than about 10% linoleic acid as percentage of total fatty acids, and a second soybean seed containing lipoxygenases 1, 2 and 3, and more than 10% of linoleic acid as a percentage of total fatty acids, which produce less than 20 cxg / g of 2 , 4-decadienal plus hexanal plus total hexanol per gram of ground soybean seeds after oxidation under mild aqueous conditions.

According to yet another aspect of the invention, there is provided a method for analyzing the odor producing property of soybean seed varieties, comprising incubating a mixture of about 1 part soy flour and about 4 parts water. for a period in the range from about 1 to about 40 minutes, and quantifying the amounts of at least one compound selected from the group consisting of 2,4-decadienal, hexanol, hexanal, and 1-octen-3-ol, and combinations of two, three or four of them, using deuterated patterns for hexanal, hexanol and decadienal.

According to yet another aspect of the invention, there is provided a method for generating soybeans comprising incubating a mixture of about 1 part soy meal or husked soybean and about 4 parts water per a period in the range from about 1 to about 40 minutes, and quantifying the amounts of at least one compound selected from the group consisting of 2,4-decadienal, hexanol, hexanal, and 1-octen-3-ol using standards deuterated for hexanal, hexanol and decadienal, and select seeds from the generated populations, based on the results.

According to another aspect of the invention, soybeans comprising a transgene are provided, such as a herbicide resistance gene conferring herbicide resistance or an insecticide gene conferring insect resistance.

According to another aspect of the invention there are provided processed foods for human consumption comprising soybeans having more than 30% protein as β-conglycinin and less than 25% protein as glycinins which produce yield less than 20 xg / g of hexane plus 2,4-decadienal plus hexanol per gram of ground soybean seeds after oxidation under mild aqueous conditions.

Description of Illustrative Modalities The invention provides soybean compositions, soybeans, and soybean seed derivatives having better organoleptic properties, and methods for producing them. The soybean compositions of the invention provide improved taste, color, odor and mouthfeel properties. The invention also provides methods for producing such compositions and methods for determining the ability of a soybean variety to produce important odors identified as 2,4-decadienal, hexanal, hexanol and 1-octen-3-hi, and the use of the results. to select seeds from generated populations.

Oxidation conditions may be produced according to the invention when approximately 0.5 part soybean flour is mixed with 2 ml water or 1 part soybean flour is mixed with 4 parts water to disperse. solid particles in water and allow oxidation reactions to take place for approximately 1 to 40 minutes at room temperature, where the ambient temperature may range from 15 ° C to 40 ° C. The concentrated suspension allows enzymes, substrates, free radicals, free radical scavenging compounds, enzyme inhibitors, and other factors to influence the amounts of odor produced.

The invention provides soya beans containing lipoxygenases and compositions derived therefrom, which have less than 4% linolenic fatty acid and / or more than 10% linoleic acid as a percentage of total fatty acids, and which produce less than 20 xg 2,4-decadienal (CH 3 (CH 2) 4 CHCHCHCHCHO, CAS # 25152-84-5) plus hexanal (CH 3 (CH 2) 4CH01 CAS n. 66-25-1) plus hexanol (CH 3 (CH 2) sOH, CAS No. 111-27-3) per gram of ground seeds after oxidation under mild aqueous conditions. The same soybeans may also produce less than 8 µg of 1-octen-3-ol (CH3 (CH2) 4CHOHCHCH2, CAS # 3391-86-4) per gram of ground seeds or husked soybean meal after oxidation under mild aqueous conditions. The 2,4-decadienal, hexanol, hexanal and 1-octen-3-ol compounds and their combinations were used to quantify the odor producing properties of soybeans. Odors are not restricted to these listed compounds. Other detectable aldehydes, ketones and alcohols may be used as measures of odor producing properties using the method of the invention. Examples of these compounds include, but are not limited to, propanal, pentenal, hexenal, pentenol, heptanal, hetenal, benzaldehyde, hexadienal, heptadienal, heptanol, octenol, octenal, nonanal, octadienone, 2-pentyl-furan, pentanal, 2,3- dimethyl-, nonenal, maltol, decenal, and 2-undecenal. According to the invention, the term "lipoxygenase" refers to an enzyme that catalyzes the oxidation of oxygen unsaturated fatty acids to produce peroxides. The term "lipoxygenase" (EC 1.13.11.12) is also referred to as lipoxidase and dioxigenase in these techniques. The odors of soy milk and soy protein ingredients from soy beans that lack one, two or three of lipoxygenases 1, 2 and 3 have been evaluated by other researchers. There are high oleic acid soybeans with less than 4% linoleic acid. Some of these soybeans that have these traits may produce less than 20 µg of the total hexanal plus 2,4-decadienal plus hexanol per gram of ground soybeans using the assay of this invention, and some will not fall within this range. . It has been found in this invention that it is possible to create soybeans containing lipoxygenases 1, 2 and 3 which produce very low odor levels, and that lipoxygenase-free soybeans can produce high levels of aroma deviations. In addition to high β-conglycinin compositions which have not been previously screened for odor producing properties, the invention particularly provides novel soybean compositions having lipoxygenases 1, 2, and 3, and more than 10% of linoleic acid. Linoleic fatty acid (18: 2 n-6) and linolenic fatty acid (18: 3 n-6) are polyunsaturated fatty acids with two or three cis double bonds according to the invention. The methods of the invention for selecting low odor producing strains from the progeny of lipoxygenase free soybeans and high oleic acid soybeans are within the scope of the invention.

1-Octen-3-ol odor is a product of the fatty acid breakdown that has hydroperoxides at the 10 carbon position of linoleic acid. It has been found in this invention that peel removal substantially reduces the 1-octen-3-ol forming property of soybean compositions. Fungal lipoxygenases and hydroperoxide lyases form 10-hydroperoxides and 1-octen-3-ol, respectively (Wurzenberger and Grosch, 1984; Husson et al., 1998). It has been found in this invention that soybeans that produce lower amounts of 1-octen-3-ol resist leakage contamination of soybean hulls, such as Phomopsis (Minor et al, 1995) and / or contain components which inhibit fungal lipoxygenases.

The organoleptic properties of soybean products depend on the content of glycine and β-conglycinin. Glycines are more susceptible to retain odors and form insoluble particles that adversely affect the sensory quality of soybean products. The present invention provides soybeans which have more than 30% protein as β-conglycinin and less than 25% protein as glycine and will produce less than 20 µg of more hexanal 2,4-decadienal plus total hexanol per gram of ground seeds after oxidation under mild aqueous conditions. The same seeds also produce less than 8 µg of 1-octen-3-ol per gram of ground seeds under similar conditions. According to the present invention, β-conglycinin refers to a protein trimer with a molecular weight of 150-200 kDa. The three major subunits are a '(72 kDa), α (68 kDa) and β (52 kDa). The major alpha and alpha subunits contain two covalently linked carbohydrate groups and the beta subunit contains one. A review of the structure and properties of β-conglycinin and the other important storage protein, glycine, can be found in Utsumi et al. (1997). A public germplasm such as "Moshidou Gong 503" can be used to change the ratio of α to β-conglycinin subunits using traditional generation methods. The term "β-conglycinin" in this patent application includes subunit variations. Seeds of the present invention which have more than 30% protein as β-conglycinin and less than 25% protein as glycine are provided, and comprise less than 5,000% per gram of free arginine, and less than 900g per gram of free asparagine. The term "free" refers to amino acids that are not bound to other molecules present in soybeans or soybean meal and can be extracted and solubilized by a 5% aqueous solution of trifluoroacetic acid (TCA) at 4 ° C. ° C all night long. The value of selecting high quality free amino acid soybeans to produce high quality soy meat, soy milk, soy flour, soy protein concentrates, and soy protein isolates had not been previously demonstrated.

The color of soy ingredients and foods according to the invention can be improved by reducing the levels of aldehydes formed (eg hexanal and 2,4-decadienal) because aldehydes react with amines to form brown pigments. . Reduced levels of lipid oxidation products may also limit the oxidative leaching of yellow pigments that make the color of the final product less white. The potential of the low leaching problem is solved in this invention by selecting soybeans that have a yellow color measured as "b * value" as used below less than 30 and which produce less than 20 0 20 of 2.4. - more hexanal decadienal plus total hexanol per gram of ground seeds after oxidation under mild aqueous conditions. The "b * value" as used herein to describe the color of soybean seed represents the CIE-L * a * b * Color Chart (CIE, Colorimetry, Publication 15.2, Second Edition, Vienna (1986), using the Colorflex procedure) and refers to the blue hues (negative numbers) to the yellow hues (positive numbers) of soybeans or soy flour; similarly, the L * value refers to the luminosity of soybeans or soy flour in the L * a * b * Color Chart. Soybeans or soya flour in one embodiment of the invention will have an L * value greater than 80.

The desired wild-type soybean, commercial cultivar, or hybrid thereof can be crossed by conventional plant breeding methods with a soybean plant which has seeds with the low odor producing phenotype of the invention to create seeds comprising the low odor trait plus other desired traits (eg yield, high β-conglycinin composition, herbicide resistance). Hybrid progeny, which have the low odor trait and other desired phenotypes, are selected. The generation methods used in accordance with the present invention include, for example, the methods described in Knowles and Briggs (1967) or any similar methods known in those techniques. Specific methods for the selection and development of new soybean varieties are described, for example, in US Patent 6,653,534.

The invention also provides processed foods for human consumption made with the soybean compositions of the invention. Examples of processed foods for human consumption may be manufactured, for example, from a peeled soy flour composition of the invention. Examples of such derivatives include, but are not limited to, alternative bars, beverages, meat and meats, soy yogurt, alternative cheeses, nutritional supplements.

The following examples are included to demonstrate preferred embodiments of the invention. Those skilled in such techniques should appreciate that the techniques described in the following examples represent techniques that the inventors have found to work well in the practice of the invention, and thus may be considered to constitute preferred modes for their practice. However, those skilled in the art should, in light of the present disclosure report, appreciate that changes may be made to the specific embodiments described and still obtain a similar or similar result without departing from the concept, spirit and scope of the invention. More specifically, it should be apparent that certain agents, which are chemically and physiologically related, may replace the agents described herein and at the same time achieve the same or similar results. All such similar substitutions and modifications, evident to those skilled in those techniques, are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims. EXAMPLE 1 Materials and Methods This example describes an analytical method of the present invention. The purpose of this analytical method is to determine the odor producing properties of different soybean strains.

[0053] The method determines the selected odors first by initiating odor formation. Grinding the seed into fine flour and activating enzymes using water caused odor formation. Studies on the rate of formation indicated that, at room temperature, odor formation was almost complete after about 20 minutes (Table 1). This time was important for the successful quantification of odorous components and evaluation of odor producing properties of different soybean strains. After 20 minutes, deuterated hexanal, hexanol and 2,4-decadienal substitutes were added to provide internal standards. The reaction was stopped by the addition of sodium sulfate, and immediately thereafter, the addition of 10% methanol in ether to extract the aldehydes, alcohols and ketones. The method was not restricted to the listed compounds. All other detectable aldehydes, ketones and alcohols can be quantified but not as accurately as the three deuterated substitutes used.

Table 1: Effect of time after mixing soy flour with water (0.5 g flour, 2 ml water, i.e. 1: 4 ratio) on the formation of aroma components. four different soybean strains (A-4, A-5, A-10 and A-14) was about 6.3.

The analytical time to determine these components in 175 samples takes 24 hours. This includes extraction and determination using gas chromatography / mass spectrometry. The sample size for analysis was typically 0.5 gram (g), but could range from 0.2 g to 0.7 g. The range of an odor concentration, on dry weight basis, was between 0.2 µg / g and 120 * g / g.

Soybean Sample Preparation: The whole seed or soybean meat was collected as samples. Approximately 6 to 12 randomly selected seeds or an equivalent weight of chunks of soybean from a sample were ground in a ball mill at a rate of approximately 1,200 revolutions per minute (rpm) for 1 minute to produce a fine flour. The seed grinding ball mill is described in US Patent Publication 2003/0146313 A1. The number of soybean seeds or meat was determined to obtain approximately 0.5 to 1.0 g of soybean meal at the end of the milling. Freshly ground soybean meal was used for further analysis.

Extraction of Soya Flour: Freshly ground soybean flour was used to extract important odors. Approximately 0.5 g (0.48 to 0.52 g) of soy flour was placed into a 20 milliliter (ml) vial (Teflon-coated transparent borosilicate sealing vials) with a cap. Demineralized water (2 mL) was added to the flour in the jar before replacing the lid. The contents of the jar were mixed for approximately 30 seconds in a swirling mixer to ensure that all flour was properly hydrated in the jar. Hydrated soy flour was allowed to incubate under mild aqueous conditions, which are defined herein as incubation in water at room temperature (22 ° C) for 20 minutes (min). After 20 min, 11 ± 0.3 g of anhydrous sodium sulfate was added to the flask, and then 10 mL of 10% methanol in ether solution was added to the flask. 30 µl of a substitute standard instillation solution (mixture of deuterated hexanal, hexanol and decadienal standards) was further added to the flask before capping again and the flask was shaken for 30-40 min in a reciprocal vessel. at -200 rpm. After 40 min, 1 mL of methanol: ether extract was placed into an Autosampler vial (Autosampler 7683 HP Autosampler vial; supplier: VWR) for further analysis.

Analysis of Soya Flour Extracts: Soybean flour extracts in methanol: ether extraction medium were further analyzed on an Agilent 6890 Gas Chromatograph (395 Agilent Technologies, Palo Alto, CA 94306, USA) equipped with Autosampler Agilent 7683 series and Leco flight time mass spectrometer with LECO Chrom TOF software (LECO Corporation, St, Joseph, M1, USA). The gas chromatograph was also equipped with a DB-WAX or DB1701 gas chromatography column with a film thickness of 0.4 or more and an internal diameter of 0.18 mm (Agilent Technologies). Methanol (methanol is purge and siphon grade EM Science) was obtained from VWR (VWR International, West Chester, PA 19380, USA); ethoxy ethane (anhydrous) was obtained from Mallinckrodt (Mallinckrodt, Hazelwood, MO 63042, USA). 2,4-decadienal, 85% trans (15% cis), hexanal 98%, hexanol 99%, 1-octen-3-ol 98%, 2-undecanone 99%, 2-nonenal 97%, and 2,4-nonadienal 99% was obtained from Sigma-Aldrich Company (Saint Louis, MO 63103, USA). Hexanal D12, Di3 hexanol and decadienal D2 were produced internally as described by Lime et al. (1999). For the analysis, 1 microliter (xL) was injected through the injection port of the gas chromatography apparatus (GC). The parameters used to analyze the samples on the chromatograph were as follows: Chromatograph Column Parameters: DB-WAX or Capillary DB 1701 10 mx 0.18 mm, 0.4 mm, Film Injection Volume: 1 µl injection coating : split / no divider coating.

Temperature Program: Initial 55 ° C for 1 min, 40 ° C to 175 ° C at 40 ° C / min, maintain 0 min 175 ° C to 240 ° C at 35 ° C / min, maintain 0 min temperature input: 220 ° C, injection mode: pulsed without division with initial 0.055 MPa (8 psi) for 1.5 min, split ratio: 20: 1. Carrier gas: 1.8 mL / min helium, constant flow.

Time Flight Time Parameters: [0059] Interface temperature: 250 ° C, source temperature: 200 ° C, mass spectrometer source temperature: 150 ° C, scan parameters: 50 to 250 m / za approximately 50 scans per second.

Analysis Quality Control: For each sample batch, a method control and instillation were performed at the same time. The instillation was done by dividing one of the samples to be analyzed in two parts. The sample should be as homogeneous as possible. The second part was instilled with a known amount of hexanal, 1-octen-3-oi and decadienal. The addition was made at the time of addition of the deuterated compounds in the extraction procedure as described above. The concentration of instilled and non-instilled compounds was determined as% recovery by the following formula:% recovery = 100 x where: Cs = instilled sample concentration at <xg / (g wet weight) C0 = sample concentration not instilled in * g / (g wet weight) wts = sample mass instilled in grams Xs = micrograms instilled in the sample The method control followed the extraction procedure without addition of soy flour.

Accuracy and Precision of Analysis: Accuracy and precision were determined using a homogeneous soybean meal sample and instilling the sample with a known level of the three compounds, hexanal, 1-oteen-3-ol and 2,4- decadiennial. Non-instilled samples were also analyzed to determine recovery amounts of the instilled compound. Different instillation levels can be used as a standard addition method to determine the basis due to systematic errors. The mean value along with the standard deviation was determined. The mean value was compared to the known material level to give an estimate of the accuracy of the method. Standard deviation gives measurement accuracy. Note that it is important to understand that these approaches only measure analytical variability, since milling 40 seeds to make soy flour homogeneous gives the average seed variability. A larger variation may occur for the actual determination because of seed variation. For hexanal, the percentage recovery was 83.8%. For 1-octen-3-ol, the% recovery was 93.6%. For 2,4-decadienal, the% recovery was 99.3%. The hexanal was the least accurate, based on the instillation approach. This is believed to be due to the volatility of hexanal and the fact that the method has low recoveries for hexanal. Octen-3-ol and 2,4-decadienal have higher degrees of accuracy using the instillation approach. Accuracy, indicated by the relative percent standard deviation of recoveries, was 5% for hexanal and e octen-3-ol. For 2,4-decadian, the precision measured by the relative percent standard deviation of the recovery values was 1.1%. Sample size has not been shown to have an effect between 0.2 and 0.9 g for this homogeneous sample, but it can be a problem between samples. This has not been tested. Therefore, the size should be about 0.5 g until the sample size proves not to be a problem or the method is adjusted.

The detection range for all compounds is between 0.5 micrograms and 25 micrograms. Adding more patterns to the standard curve can widen the range. EXAMPLE 2 Identification and Selection of Low Odor Producing Soya Beans Potent odors were quantified in soy milk made from a control soybean (Vintor 81), a lipoxygenase-2-free soybean (QT). -1) and a soybean grain without lipoxygenases 1, 2 and 3 (IA2Q25). Soymilk was made from each soybean variety by soaking the desired clean seeds in water at a 1: 5 ratio (1 gram weight: 5 mL water) at 25 ° C for about eight hours. After discarding the soak water, soybeans weighing twice the original weight were drained and mixed with fresh distilled water (2 x the dry weight of soybeans) for five minutes. Further distilled water (7 x the dry weight of soybeans) was added to the slurry for about 2 min at 20 ° C. The slurry was slowly baked at 95 ° C to 98 ° C for 20 min in a water bath, and filtered through a thick cotton cloth and squeezed manually to extract as much soy milk as possible. Soymilk was pasteurized by baking in a water bath at a temperature between 85 0 and 90 ° C for 10 min to reduce microbial contamination, and stored at 4 ° C before odor extraction for further analysis.

The odors were extracted from the soy milk samples. Soymilk was extracted with a portion of Freon® 113 for at least 30 min. After removal of Freonâ „¢ 113 extract, the aqueous phase was further extracted with 0.67 part ethyl acetate. After collecting the ethyl acetate extract, the aqueous phase was discarded. Feon ™ and ethyl acetate were filtered through magnesium sulfate to remove as much water as possible and concentrated to 1 mL using a Buchi 0.1 rotary evaporator. Freon extracts were evaporated under 48 kilopascals (kPa) and ethyl acetate under 86 kPa.

Potent odors in soybean milks were quantified using GC olfactometry (Acree, T.E., Analytical Chem. 69: 170A-175A (1997)). GC olfactometry is a gas chromatography with an inhaler orifice, where the potency of a chemical compound as an odorant is measured as a measure of the human response to odorants in an air stream or puff. A Hewlett Packard 6890 gas chromatograph equipped with a methyl silica cross-linked fumed silica column (film thickness = 0.33 µm) was used for CharmAnalysis® analysis (Acree, TE, Bamard, J., Cunningham, DG Food Chem 14: 273-286 (1984)). The effluent consisted of helium (2 mL / min) as carrier gas and nitrogen as replacement gas (about 30 mL / min). The effluent was mixed with the sniff air (20 mL / min) which was 99% humidified laboratory air to between 50% and 75% and passed through the inhaler through a 10 mm diameter silylated Pyrax tube. The GCMS furnace has been programmed to begin raising its temperature three minutes from the initial temperature of 35 ° C to 225 ° C at 6 ° C / min. Further details of the CharmAnalysis ™ method in soymilk can be found in the Ph.D. dissertation presented to the faculty of the Graduate School of Cornell University by Yu-Wen Feng.

The two lipoxygenase-2-free soybeans (IA2025, QT-1) produced lower hexanal levels than the control (Table 2). Soybean grain without all three lipoxygenases (IA2025) produced the highest levels of 2,4-decadienal and 1-octen-3-one, while soybean without lipoxygenase-2 (variety QT-1) had the highest levels. lowest levels of all five potent odors (Table 2). It was evident from the results that unknown compositional factors other than lipoxygenase-2 were involved in the control of lipid oxidation in soybeans. The QT-1 soybean variety has been identified as a useful variety for creating low odor commercial soybean varieties, whether or not lipoxygenase-2. The method of Example 1 was developed to identify progeny of QT-1 and other soybean strains that produce low amounts of 2,4-decadienal, hexanal, hexanol and 1-oten-3-ol.

Table 2: Attractive value of odors present in soy milk prepared from three soybean varieties. The Monsanto strain produced lower levels of aroma deviations than one soybean-free lipoxygenase (IA2025) and one tofu-soybean grain (Vínton 81).

Table 3 describes the soybean crosses that were used to develop progeny to describe the invention. Standardized methods were used to generate these strains. Table 3: Intersections of Soybeans Used to Develop Progeny Used to Describe Soybeans and Methods of Invention EXAMPLE 3 Demonstration of Ear-to-Ear and Local-to-Local Odor-producing Characteristics Low Amino Acid Content of Soybean Strains Selected According to the Invention [0069] For color evaluation, seeds were collected as samples. The desired number of seeds from a sample was ground in a Mega-Grinder at a speed of 1,200 rpm to produce a finely ground soybean meal. Mega-Grinder for grinding seeds is described in US Patent Publication No. 2003/0146313 A1. Freshly ground soybean meal was used for further analysis.

The ColorFlex Model 45/0 Spectrocolorimeter Color Measurement System manufactured by Hunter Laboratories (Hunter Associates Laboratory, Inc., Reston, VA, USA) was used to measure the color of soy flour by the operating procedure suggested by the manufacturer. . Colors were measured on the CIE-L * a * b * Color Chart (CIE, Colorimetry, Publication 15.2, Second Edition, Vienna (1986), using the Colorflex procedure. The International Commission on Lighting, abbreviated CIE, by its French title Commission International de l'Eclairage, and an organization dedicated to international cooperation and information exchange among its member countries on all issues related to lighting science and technique.The L * value refers to the brightness of soy flour while that the b * value refers to the blue (negative numbers) to yellow (positive numbers) of soy flour. The color values of soy flour made from different strains are shown in Tables 4 and 5.

Free Amino Acids: [0071] Unground samples were stocked in an isolated room with controlled temperature and humidity (approved by APHIS, Animal Health Inspection Service and Veaetalt. The samples were ground using the Mega-Grinder CAT to create flour. stored at 4 ° C in seed storage salt The soybean meal was extracted with 5% TGA at 4 ° C overnight, centrifuged, and the extracts stored at -80 ° C. filtered, diluted if necessary, and analyzed for free amino acids by the OPA method The OPA method uses o-phthalialdehyde (OPA) to derive samples prior to injection into a reverse phase HPLC C18 column.The derived primary amino acids are efficiently separated per group R and quantitatively detected by a sensitive fluorometer.The relative standard deviation for this method is -3%, Ypoxygenase activity: The samples were ground using a Mega-Grinder. The freshly ground meal was weighed in triplicate (5 ± 1 mg) and placed into a specific well of a 2-well 96-well extraction plate. Samples were extracted with 0.1 M K 2 PO 4 (pH 7.0 or pH 9) for 1 hour at room temperature. After centrifugation, the obtained supernatant was used to measure linoleic acid (substrate) consumption using a spectrophotometer and then determination of total protein per sample using a Βίο Rad Protein Dye. The unit of lipoxygenases was calculated using absorbance changes at 230 nm during a 1 minute reaction and using the extinction coefficient (ε = 23,000 M'1cm1). The substrate concentration consumed during the reaction was calculated by substituting each value in equation A = ebC. One unit of lipoxygenase was defined as the substrate micromoles consumed per minute and total mg of extracted protein. Using reagent solutions prepared at pH 7.0 or pH 9.0, this assay allows to measure the levels (Enzyme Units) of lipoxygenase-2 / -3 or lipoxygenase-1, respectively. Results of lipoxygenase units are provided in LOX Units, pH 7.0, for lipoxygenase-2 and 3 activities and as LOX units, pH 9.0, for lipoxygenase-1 activity.

Results: The odor characteristics of soybeans persisted when cultivated in multiple locations (Table 4) and multiple years (Tables 4 and 5). Soybean strains that produced low levels of hexanal, hexanoi and 2,4-decadienal and 1-octen-3-ol in the odor assay (described in Example 1) were consistently different from strains that produced high levels (Table 4 ). The soybean strains in Table 4 were selected in order of the levels of hexanal + hexanoi _2,4-decadienal that the strains produced. For example, strain A-1 at the top of the table produced 18.21 ± 4.21 µg / g of the three odors. The amounts of hexanal plus hexanoi produced by the same crossover soybeans (such as crossover A) correlated with the amounts of 2,4-decadienal produced (Table 6, R2 = 0.85}, suggesting similar mechanisms. and control by genetic and compositional variations.The levels of 1-octen-3-ol produced were independent of hexanal, hexanoi and 2,4-decadienal (Table 6, R2 = 0.01), suggesting a different mechanism of formation and control. Soybean strains grown at 2-3 sites producing low levels of 1-octen-3-ol were consistently different from strains producing high levels of 1-octen-3-ol. 18 produced 4.70 ± 0.88 µg / g of 1-octen-3-o and lineage A-12 produced 14.67 ± 2.37 µg / g of octen-3-ol. which have a combination of genetic compositions that produce lower levels of 1-octen-3-ol and lower levels of more hexanal, more hexan decadienal hi (such as lineage A-6).

Tables 4A and B: Color and odors produced by ground soybeans produced from progeny of crosses A, B, C, D and E, as shown in Table 3. Values are mean and standard deviation (Stdev). for each strain grown at two or three sites in 2002 (Ames, Iowa; Oxford, Indiana; Gladbrook, Iowa). The progeny of the listed B cross had a low iinolenic acid content of 2.9 ± 0.4% of total fatty acids. All strains have yellow hilum. The ground soybeans had a moisture content of 8%. The color values are L * (luminosity), a * (green-red) and b * (blue-yellow). Abbreviations: Stdev = standard deviation.

Tables 5A and B: Characteristics of soybean meal made from soybean, commercial commodity (controls) and progeny of crossings A, B, C, D and E grown in 2001 Soybeans ground had a moisture content of 7%. The order of the strains is the same as in Table 4. The progeny of the crossing B listed had a trace of low linolenic acid content (2.9 ± 0.4% of total fatty acids). Two progenies lacked one or more lipoxygenases, the others contained all three lipoxygenases designated L1, L2 and L3. Lipoxygenase activities have substrate micromole units consumed per mg substrate. The color values are L * (brightness), a * (green-red) and b * (blue-yellow). Abbreviations: Stdev = standard deviation.

Table 6: Linear relationships, determined by R squared values, between odors produced from soybean progeny harvests of 2001 and 2002. Linear regressions were calculated from the data in Tables 4 and 5. Abbreviations: Η + H = hexanal + hexanol; D = 2,4-decadienal; O = 1-octen-3-ol; DHH = 2,4-decadienal + hexanal + hexanol.

It was possible to develop low odor, low color varieties with good yield for the first time using the methods and soybeans of this invention. For example, strains A-1, A-4 and A-6 produced 90, 90.5 and 104% of those of commercial controls.

There was little or no relationship between lipoxygenase activities at pH 7 and 9 and the formation of 2,4-decadienal + hexanal + hexanol from hydrated soybean meal (R squared values <0.35, Table 7). Soybeans that produced significant lipoxygenase activities at pH 7 and 9 (strain A-1) had similar odor properties to the strain lacking lipoxygenase activity (strain C-1) (Table 5). Commercially available soybean, which lacks lipoxygenase activities 1, 2 and 3 (IA2032 from the 1999 harvest) was also tested for lipoxygenase activity and odor formation. No lipoxygenase activity was found at pH 7 and 9 for soybean meal free of the three lipoxygenases, but significant levels of hexanal (23.3 * g / g), hexanol (14.9 <xg / g) and 2.4 decadienal (5.8 * g / g) were formed in the odor test of the invention. After experiments using lipoxygenase inhibitors, it was concluded that at least one other lipoxygenase was active in this soybean composition.

Light colored protein ingredients have been appreciated by the food industry, especially for dairy products. Soybeans were grown at 2-3 sites to determine if low-colored soybeans could be selected. It was found that low color soybeans could be selected as determined from the b * value of ground soybeans (high b * value indicates more yellow, less blue; Table 4). For example, A-2 soybean had a b * value of 22.16 ± 1.39 and A-6 soybean had a b * value of 27.35 ± 2.85 (Table 4). There was a correlation between the b * value of soybean lines cultivated in 2001 and 2002 (R squared = 0.7; Table 7).

Table 7 Linear relationships determined by R squared values for odor, free amino acid, and color, produced from soybean progeny harvested in 2001 and 2002. Linear regressions were calculated from the data in Tables 4 and 5. The free amino acid correlations with the 2002 data were made without three out-of-bound lines (A-4, B-5 and B-6). Environmental factors, besides genetic, affect the compositions that influence the odor-producing properties of soybeans. The environmental effects were evident comparing the odor-producing properties of soybean strains grown in two seasons, 2001 and 2002. The range of 2,4-decadienal + hexanal + hexanol for the strains raised was 12-44 ocg / g. in 2001 and 17-65 xg / g in 2002 (Tables 4 and 5). The producing properties of 1-octen-3-ol appeared to be more sensitive to environmental factors, as evidenced by the lack of correlation between 1-octen-3-ol produced by the same strains cultivated in 2001 and 2002 (R squared = 0.62). , Table 6).

The amounts of free arginine (Arg) and free asparagine (Asp) in soybean progeny were determined. The amounts of arginine and asparagine correlated (R2 = 0.81, Table 7) and the arginine plus total asparagine in soybean did not correlate with the selected odor and color properties of soybean (R squared <0, 3, Table 7); therefore, free amino acid testing is required to select strains that have low 2,4-decadienal, low color, and low amino acid combinations. The feasibility of selecting low amino acid soybean strains was based on the good correlation between free arginine + asparagine in strains harvested in two different years (R squared = 0.8; Table 7). EXAMPLE 4 Combination of Low Odor Producing Property with High β-Conglycinin Composition and Low Arginine and Asparagine Free Compositions and Low Color Property The origin of the high β-conglycinin trace was a mutated soybean. lacking glycinins and containing about 55% of the total protein as β-conglycinins (US Patent 6,171,640). A lipoxygenase assay was not useful for selecting low odor producing strains as described above. Soybeans were created that had protein, fat and amino acid spectra that were within normal ranges for commodity soya beans. Quantification of Soybean Protein Subunits: About 8 seeds were ground using a Mega Grinder (US Patent Publication No. 6,171,640). For each sample, ~ 30 mg of flour was extracted into 1.0 mL of SDS Laemmli Buffer pH 6.8 with 0.1 M DTT in a rotary or multi-plate vortex for 45 to 60 min. The tubes were centrifuged for 3 to 5 min. A portion of the supernatant was transferred by microcentrifugation tubes and diluted with the above buffer to yield 1.2 to 1.5 cg / dm of total protein. The samples were boiled for 3 min, cooled and centrifuged. Pre-cast Criterion gels, with 102-% Tris-HCI gradient, were loaded with 15-20 æg protein from each sample. The gels were electrophoresed at 180-200 V in 1X Tris-Glycine-SDS Buffer until the tracking dye reached the bottom of the gel for about 1.2 h. The gels were fixed for 30-60 min in 40% methanol / 10% acetic acid, and stained with Coomassie Colloidal Blue G-250; at least all night long or for up to 3 days. To remove the bottom, the gels were discolored with demineralized water. The gels were imaged using the GS 800 Calibrated Densitometer. Quantitation was performed using Bio-Rad Quantity One software. The software is used to determine the relative amount of each band in the sample row. Percent glycine subunits and percent β-conglycinin subunits were reported as a relative percentage of total protein in the row.

Total Amino Acid Analysis: The sample was tested by three methods to obtain the complete profile. Tryptophan required basic hydrolysis with sodium hydroxide. Sulfur-containing amino acids required oxidation with performic acid prior to hydrolysis with hydrochloric acid. Analysis of the samples for remaining amino acids was performed by direct acid hydrolysis with hydrochloric acid. Once hydrolysed, the individual amino acids were then quantified using an automated amino acid analyzer (AOAC International Official Methods of Analysis, 2000).

Ash: The sample was placed in an electric oven at 550 ° C and ignited to expel all volatile organic materials. The remaining nonvolatile matter was quantified by gravimetry and calculated to determine the ash percentage. (AOAC International Official Methods of Analysis, 2000).

Carbohydrates: Total carbohydrate level was calculated by difference using data derived from newly obtained weight and the following equation:% carbohydrates = 100% - (% protein +% fat +% moisture +% ash) . (United States Department of Agriculture, 1973).

Soxhlet Extraction Fat The sample was weighed into a cellulose thimble, containing sand and sodium sulfate, and dried to remove excess moisture. The pentane was dripped through the sample to remove grease. The extract was then evaporated, dried, and weighed. (AOAC International Official Methods of Analysis, 2000).

Humidity: The sample was dried in a vacuum oven at approximately 100 ° C to constant weight. Moisture weight loss was determined and converted to moisture percentage.

Protein: Nitrogen compounds in the sample were reduced in the presence of fuming sulfuric acid and a mixture of mercury catalysts to form ammonia. Acid digestion was made alkaline. Ammonia was distilled off and then titrated with a standard acid. Nitrogen percentage was calculated and converted to protein using factor 6.25. {AOAC International Official Methods of Analysis, 2000; Bra d Street (1965); Kalthoff and Sandell (1948)).

Results: A population of high β-conglycinin soybean strains selected from a cross with a soybean that contains the low odor producing trait showed a wide range of odor producing properties, measured by the formation of hexanal, hexanol, 2,4-decadienal and 1-octen-3-ol (Table 8). Commodity soybeans contained about 22% of the total protein as β-conglycinins and about 38% of glycines, compared with the soybeans in Table 8 that had more than 30% protein as β-conglycinins and less than 25% protein as glycine. Soybeans having more than 30% of the total protein as β-conglycinins were produced, which produced less than 20 µg / g total of hexanal, plus hexanol and 2,4-decadienal in the odor test of Example 1, and also comprise low levels of free asparagine and arginine (Table 8). For example, 20 high β-conglycinin strains were created by the invention, which produced less than 20 µg / g of the total plus hexanal 2,4-decadienal plus hexanol and a total free arginine plus asparagine between 360 and 2,840 µg / g ground soybeans (Table 8). The free asparagine of these strains was between 35 and 1,000 cg / g and the free arginine of these strains was between 500 and 2,400 cg / g of ground soybeans.

Tables 8A and B

Progeny characteristics of high β-conglycinin soybeans harvested in the United States. Soybeans have been raised which have more than 30% protein as β-conglycinins and less than 25% protein as glycines, which also produce less than 20 = cg / g of more hexanal 2,4-decadienal, more hexanol per gram of soy flour in the odor assay of Example 1. The lipoxygenase activity assay was not useful for identifying low odor producing strains.

Table 8A

Lipoxygenase activities varied significantly in high β-conglycinin soybean samples. There was no relationship between lipoxygenase activities at pH 7 and 9 and the total amounts of 2,4-decadienal plus hexanal plus hexanol produced from the same soybean meal (R2 values <0.02).

An additional experiment was conducted to demonstrate that soybeans that have more than 30% β-conglycinin and less than 25% glycinins that have low color and an amino acid composition that is within the range of the grains can be selected. soybean commodities. Four soybeans with a high β-conglycinin content were selected, which also contained the Roundup Ready® trait, which produced the same or better than average commercial quality requirements at three sites. These soybeans had the color trait illustrated by a b * average of about 22 and an L * average of about 85 (Table 9). Amino acid compositions (Table 9) fell within the ranges of export soybeans (Table 10), as published in the Life Science Institute Crop Composition Database (version 1.0, accessed March 22, 2004). The average amino acid compositions of the three high β-conglycinin soybean strains (Table 9) were compared with the average soybean composition in the ILSI database (Table 10). Four amino acids (arginine, lysine, histidine and serine) from the high β-conglycinin strains were between 10 and 15% different from the average composition of export soybeans, but were still within the range of soybean grain compositions. export.

It is also anticipated that the organoleptic properties of high β-conglycinin soybeans will be further improved by crossing with low linolenic and medium oleic soybeans. Monsanto's low-linolenic and medium-oleic soybeans have been grown using traditional generation and contain a linolenic content of about 2%, a linolenic content of about 25% and an oleic content of about 59% of the acids. total fat.

Table 9 Composition »color and odor properties of soybeans grown at three sites in 2003. All had the Roundup Ready" trait and dark hilum.

Table 10 Comparison of the Average Composition of the Three High Beta-Conglycinin Soybeans with the Average Soybean Composition in the Life Sciences Institute Crop Compositions Database (version 1.0). Selection criteria used to obtain the data: crop type: soybeans Glycine Max; tissue type: seed; crop year, everything, country, everything, state, everything. Abbreviations: FW = weight of soy flour; DW = dry weight; HBC = high beta-conglidin content EXAMPLE 5 Comparison of Export Soybean Odor and Color Properties with Soybeans Selected According to the Invention The odor and color properties of export soybeans were determined to compare with the odor and color properties of the soybeans of the invention. Some soybeans produced less than 17.5 µg / g hexanal plus haxanol and some produced less than 11 µg / g 2,4-decadienal in the odor test (Table 11). However, none of the exported soybeans produced less than 20g of decadienal plus hexanal plus hexanol per gram of ground seeds after oxidation under mild aqueous conditions. Four lipoxygenase-containing strains of this invention were created which produced less than 20 µg / g hexanal + hexanol and 2,4-decadienal in two harvests (Tables 4,5). For example, lineage A-1 from the 2001 and 2002 harvests yielded 18.0 and 18.2 ocg / g, respectively, of the sum of hexanal + hexanol and 2,4-decadienal (Tables 4, 5). More than 20 high β-conglycinin strains were created by the invention, which produced less than 20 µg / g hexanal + hexanol and 3.2 µg / g 2,4-decadienal in two crops (Table 8). . The color of export soybeans covered a wide range. For example, the value of b * was in the range of 27 to 34 (Table 11). The soybean strains of the invention have been shown to extend b * values as low as 22 (Table 4, Table 9). EXAMPLE 6 Demonstration of Ability to Combine Low Odor Producing Trace with a Low Linolenic Acid Composition Trace This example demonstrates that it is possible to combine the low odor producing trait with a low linolenic acid composition trait, and It also explores the effects of peeling soybeans on odor production. The percentage of soybean fatty acids that is linolenic acid is usually about 8%. Using traditional generation, it was possible to create soybeans that have less than 6% linolenic acid and low odor producing property (Table 12). The formation of 1-octen-3-ol is independent of the formation of other volatile compounds measured in the odor test. One hypothesis is that fungal enzymes on the surface of soybeans are the source of rapid 1-octen-3-ol formation in the assay. It has been theorized that peeling the seed before grinding to flour should lower the levels of fungal enzymes in the odor test. Whole soybeans are usually ground to produce the flour that is used for odor testing. A new test was performed using six low odor, low linolenic soybean strains, which were carefully peeled. The amount of 1-octen-3-ol formed from peeled seeds was about half that of whole seeds, supporting the hypothesis that components such as fungi and fungal enzymes in the shell part contribute to 1-octen-3-ol formation (Table 13). Lipoxygenase-free soya beans often produce higher levels of 1-octen-3-ol. It has been theorized that lipoxygenases may play an important role in inhibiting mold growth, and therefore, in the absence of lipoxygenases, mold infections may be greater, allowing more fungal enzymes and 1-octen-3-ol formation. . Low-odor soybeans containing lipoxygenases that have been identified by the screening of this invention tend to have a lower 1-octen-3-ol content than higher-odor soy beans (as per Table 12).

Tables 12 A1 to A4: Color-producing and Odor-producing Properties of Low Linolenic Soybean Strains Color values are L * (lightness), a * (green-red), and b * (blue-yellow). The formation of 1-octen-3-ol in the odor test was reduced when soybeans were husked before making soybean flour.

Effects of pH on Odor Formation A standardized odor test provided by the invention involved the addition of soy flour to water, resulting in the formation of odors at about pH 6.3. The purpose of the following experiment was to determine whether varieties that produce low odor levels under this condition also produce low odor levels under other pH conditions. The odor producing properties of a commercial control soybean were compared with a low odor producing strain (A-4). The A-4 strain produced lower decadienal and hexanal levels at pH 3.0 and pH 5.5, and pH 7 and pH 9.2 (Table 14). These data support the use of the bufferless assay as a method for Select soybeans that will produce lower odor levels within a wide range of pH conditions. The highest concentrations of hexanal and 2,4-decadienal were produced at pH 9, the pH at which the smallest amount of 1-octen-3-ol was produced (Table 14).

Table 14 Effects of pH on odor formation from commercial control soybean and low odor producer soybean (A-4 of Figures 4 and 5, harvested in 2003). The use of the odor test for this measurement was the same as for Example 1 except that soy flour was added to 0.1 M K2HP04 at its pH (3.02, 5.45, 7.01 and 9.16). . EXAMPLE 8 Effects of Soybean Protein Composition on the Soymilk Sediment This example describes the lower levels of the sediment formed in soybean milk made from soybeans having the modified protein composition of the invention. Protein-containing sediment has a negative impact on the organoleptic quality of soy milk, as no particulate matter is desired in beverages.

Soymilk Preparation: A low-odor control soybean (A-4) and a soybean that has about 39% β-congicinin and about 13% glycine were ground using a Mega Grinder to manufacture soy flour. Each flour sample was added to 36.75 g of water (4 ° C) in a disposable polypropylene centrifuge tube such that the final protein concentration in the mixtures was 3.3 wt%, and passed through. 15 seconds with an output power control setting of 8. The samples passed on the ultrasound were centrifuged for 10 min at 8,000 rpm at 4 ° C using Eppendorf 5804 R Centrifuge, Supernatants soybean) were decanted into disposable polypropylene centrifuge tubes. A portion of each soy milk (27.75 g) was transferred to another 50 ml disposable centrifuge tube. The above samples were prepared in duplicate to allow for variation in the following sucrose addition. Sac rose (0.6987 g) was added to 27.25 g of soy milk before or after heat treatment.

Soymilk samples were heat treated by placing the 50 ml disposable polypropylene centrifuge tube into a 95 ° C silicone oil bath for 5 min, and then the samples were transferred to an ice bath to cool, and then stored in a refrigerator for 30 days. A sediment formed in the soy milk samples over time. The amount of sediment formed was quantified as follows. The sediment at the bottom of the centrifuge tubes was dispersed by turning the tubes back and forth. Soymilk samples were transferred into a tared centrifuge tube and the final weight was recorded. The tubes were placed into the Eppendorf 5804 R Centrifuge and centrifuged for 2 min at 8,000 rpm and 25 ° C. The amount of soy milk was decanted and the weight of the remaining sediment was calculated. The amount of sediment was recorded as a percentage of soybean milk weight (% of sediment = 100 x Sediment weight / Soymilk weight).

Results: Control soybean milks had at least twice the sediment content of high β-conglycinin soy milks (Table 15).

Table 15 Effects of Soybean Protein Composition on Soymilk Sediments, A-4 is the control soybean, HBC is a high β-conglycinin and low glycinin soybean.

The above examples indicate that unique compositions have been created that can produce low odor levels even when soybeans contained lipoxygenases 1, 2 and 3. The ability to select soybeans which have better organoleptic properties was also demonstrated where Soybeans are selected based on the amounts of free glycine, arginine and asparagine proteins, yellow pigments and polyunsaturated fatty acids in the soybeans, and the amounts of the 2,4-decadienal, hexanal, hexanol and 1-octen-3-ol odors. , produced by aqueous suspensions of ground soybeans. A method for estimating the potential for soybeans to produce 2,4-decadienal, hexanal, hexanol and 1-octen-3-ol, and other even more potent odors in soybean ingredients and foods has been described. In this method of estimating soybean odors, the importance of incubating soybean flour in water at a 1 to 4 ratio resulted in peculiar observations that 2,4-decadienal forms rapidly at room temperature in suspensions, and It is possible to generate soybeans containing lipoxygenases 1, 2 and 3 which produce very low levels of odors.

It has also been shown in the examples that it is possible to create endogenous soybean compositions having more than 30% β-conglycinins and less than 25% glycine, which have normal or low levels of free arginine and free asparagine. It has also been shown that the low odor and color properties can be combined with high β-conglycinin compositions, and other combinations with soybeans having low linolenic acid and medium oleic acid content have been designed. Glycines are a source of insoluble protein in soy foods and ingredients, creating sediment in beverages and an undesirable mouthfeel. Free arginine and asparagine may form ammonia during processing, which may further react with odor compounds such as 2,4-decadienal to form potent odors such as 2-pentyl pyridine. Linoleic and linolenic acids are polyunsaturated fatty acids that are substrates for odor formation; Lowering your soybean content will help lower odor formation. Soybean pigments contribute to color deviations of soybean products, further impacting the organoleptic response. All in all, the high β-conglycinin soybean composition, low free arginine and asparagine content, low odor and low color, and low polyunsaturated fatty acid content, and the most valuable composition of the invention to create protein ingredients and soy foods that are organoleptically pleasing. It is also recognized that these compositions will not lack healthy properties associated with soy protein ingredients, as β-congenins are associated with soy protein cholesterol and triglyceride lowering properties (Duranti et al, 2004) and inhibition of soybean protein. arteriosclerosis (Adams et al., 2004). EXAMPLE 9 Additional Sedimentation Analysis of Low Odor Soy Compositions The study in Example 8 was repeated with the following changes. One export soybean was included as a control (AG3302). The soybeans were peeled before making the flour and the flour was sieved before addition to water. Supernatants (soymilk) were transferred to tared centrifuge tubes. Sucrose was added prior to heat treatment to produce a final concentration of sucrose of 2.5% by weight. The samples were stored in a refrigerator for 21 days. The heights of the soy milk pellets in the centrifuge tubes were measured, and then the samples were centrifuged for 5 min at 8,000 rpm. The supernatants were decanted and weighed, the pH of the supernatants determined (all samples had pH = 7), and the tubes containing pellets were weighed. The percentage weight of wet soybean sediment was calculated for each sample (% weight of wet soybean sediment = 100 x weight of wet soybean sediment / (weight of wet soybean sediment + weight of soybean supernatant)).

The beneficial effects of low odor HBC soybeans in reducing protein sediment formation were obtained using peeled soybeans (Table 16). HBC soybean milk had a 2.2 fold reduction in sediment compared to the low odor soybean lineage and a 7 fold reduction in sediment compared with the control soybean. A surprising 3.1-fold reduction was found in sediment in low-odor soybean compared to control. It is possible that the low odor trace reduced sediment formation by limiting free radical formation and protein oxidation. This therefore indicated the additional beneficial properties of the optimized composition by combining high β-conglidnine and low odor traces.

Table 16 Effects of high-odor, low-beta-conglycinin traces on Soymilk Sediment Height and Sediment Pellet Weight Compared to a Control Soybean Composition EXAMPLE 10 Preparation of Soybean Flour Peeled Soia and Soy Protein Isolate Ingredient A control soybean (AG3302) and soybeans comprising more than 30% of total proteins as beta-conglycines and less than 25% of total proteins as glycinins and less than 2,500 micrograms / gram free arginine plus asparagine (DJB2104GOR, EXP319AP) were husked to make husked soybean meal, and then further processed to make isolated soy protein ingredient according to the steps below.

Table 17 Composition of beta-conglycinins and glycinins in different soybean strains. 1. Adjust soybeans to about 10% humidity and room temperature. 2. Fractionate soybeans using a fractionation mill. 3. Peel the fractionated soybeans using a vacuum cleaner. 4. Condition the split and peeled soy beans at 50-60 ° C using a cooker. 5. Flock conditioned soybeans using a Flocking Mill. 6. Extract the soybean flakes with hexane. 7. Remove solvent from defatted soybean porridge, resulting in flakes. 8. Grind the flakes to make flour. 9. Add water to a jacketed 300 L tank and adjust to 50 ° C and pH 9.0 using 40% NaOH. The defatted soy flour was added and mixed. The ratio of water to soy flour was 12: 1 (by weight). The extraction time was 45 min. 10. Recover solubilized soy protein from the extraction slurry using a mud-removing disc centrifuge (0.4-0.414 MPa (58-60 psi) back pressure). 11. Adjust the pH of the clarified protein solution to 4.5 by adding hydrochloric acid (18%) and allowing to react for 30 min at 45 ° C. 12. Recover the precipitated protein using a mud-removing disc centrifuge. 13. Wash protein clot twice using acidified water (pH 4.5 ± 0.1, 30-35 ° C). The wash water to compressed wet solids ratio was 6: 1 by weight. The protein clot was recovered using a mud-removed disc centrifuge after each wash (0.4-0.414 MPa (58-60 psi) back pressure). 14. Mix protein clot with sodium hydroxide (30%) to adjust pH to 6.8 using 30% NaOH and then heat treat at 116 ° C for 7.5 seconds. Then adjust the pH to pH 6.8. 15. Adjust the temperature of the protein solution to 45-55 ° C and atomize using an inlet air temperature of 204-215 ° C and an outlet air temperature of 82-88 ° C. 16. Determine the nitrogen solubility index of isolated soy protein ingredients. Suspend a portion of the sample in water under stirring at 30 ° C for two hours. Then dilute with water to a known volume. Analyze a separate portion of the sample for total protein and analyze an aliquot for protein by the same method. Calculate water-soluble protein as a percentage of total protein, which was proportional to water-soluble nitrogen as a percentage of total nitrogen. Results: The nitrogen solubility index of soy protein powders (Table 18) was directly proportional to the amounts of beta-conglycinin in soybean grains used to make soy protein ingredients: NSI = 1.4716 (% of beta congliclnl) + 7.4502; R squared = 0.9975. A reduction in insoluble protein levels improves the organoleptic quality (more pleasant and refreshing sensation) of food products formulated with soy protein ingredients.

Table 18 Nitrogen Solubility Index The amino acid composition of isolated soy protein ingredients made from soybeans was similar, except that the lysine content of the high beta-conglycinin (HBC) ingredients it was 6% higher than the control.

Table 19 Total Amino Acid Composition of Soy Protein Ingredients EXAMPLE 11 Preparation of Soybean Product Grown From Blanched Soybean Meal Blanched soybean meal was prepared from an export soybean (AG3302), a high beta-conglycinin soybean (DJB2104GOR) and a low odor producing soybean strain (03JBK8-25), all harvested in the United States in 2004. The methods used to prepare and test the cultivated product of the Test made from soybeans are: 1. The soybeans were fractionated using a fractionation mill. 2. The broken soybeans were peeled using a vacuum cleaner. 3. The soybean (peeled soybean) meat was ground by 1 pass through a hammer mill and 5 passes through a pin mill. 4. Peeled soybean meal was packed in plastic bags inside fiber drums. 5. The protein content of the flour was determined. 6. The flour from the peeled soybeans (3 ° C) was added to the water (3 ° C) such that the protein content was 3.5% (weight basis) and mixed using a homogenizer. portable for about 1 to 2 min. 7. The soybean meal suspension was heated in a plate heat exchanger, then steam injected to treat the suspension at 141 ° C for 3.5 seconds, and deaerated before cooling to about 4 ° C. C, 8. The heat treated suspensions were filtered to remove the fibers, resulting in soy milk. 9. Dairy flavored ingredients, sugar (3%) and salt (0.2%) were added to soy milk and mixed using a portable homogenizer. 10. Flavored soymilk was heated in a plate heat exchanger and then steam injected to treat the suspension at 141 ° C for 3.5 seconds, and deaerated before cooling to about 4 ° C. , and packed in sterile containers. 11. Soymilk samples, each containing 2.2% protein, were weighed into 1 liter sterile vials and heated in the microwave oven for 45 to 60 seconds (about 24 ° C). 12. Sugar (3.1%), vanilla extract (0.4%) and cultivated soy yogurt (L. bulgaricus, S. thermophillus, L. acidophilus, B. bifidum, L. casei, L. rhamnosus (8%) were added to soy milk and the samples were mixed 13. The culture containing soy milk was placed in a 43 ° C incubator and incubated for 4 hours, removed and refrigerated overnight (4 ° C). PH and viscosity measurements were made on refrigerated samples and a sensory evaluation was conducted by a panel of 3 people who did not know the sample formulations.The viscosity was measured on a Brookfield Viscometer with Rod # 3 at 20 rpm.

Results: The aroma profile of the cultivated product made from DJB2104GOR was pleasant and would suit a fruit flavored drink. The product profile made from 03JBK8-25 was pleasant and would fit a curd or paste. It has been theorized that a combination of high beta-conglycinin and low odor producing traits would also create pleasant cultivated soy milk products. The lower viscosity of high beta-conglycinin material can help to create higher protein content products at the same consistency level.

Table 21: Taste Profile of Cultivated Product EXAMPLE 11 Demonstration of Low Odor High-β-Conglycinin Combination This example describes the combination of compositions with high β-conglycinin low-odor producing properties. It demonstrates the ability to produce low odor soybean compositions comprising lower glycine content and higher β-conglycinin content. The types of crosses, such as A and E (Table 3), were combined with the high β-conglycinin germplasm, which has a pedigree of A3233 / B2G2 / A1923. Protein analysis was conducted as follows: Eight soybeans were selected and ground using the Mega-Grinder CAT (SOP Asci-01-0002). The ground samples were stored at 4 ° C. For the analysis, ~ 30 gm of flour each was weighed into a 96-well 2 ml microtiter plate. The protein was extracted for 1 hr under 1 ml of 1X SDS Laemmli pH 6.8 buffer containing 0.1 M dithiothreitol (DTT) as a reluctant. After centrifugation, part of each extract was further diluted in SDS buffer to yield 0.2-0.5 æg / æl total protein, heated to 90-100 ° C for 10 min, and cooled. For each sample, 1-2 æg of total protein was loaded using a 12-channel pipette onto a 26-row 15% T-gradient Tris / HCI Criterion gel. Molecular weight standards and parental control were included in two of the rows on each gel. The gels were electrophoresed until the tracking dye reached the bottom of the gel within ~ 1.2 hours, then stained overnight in Colloidal Coomassie Blue G-250, bleached in demineralized water, and reproduced using the GS800 Calibrated Densitometer. Quantitation was performed using Bio-Rad Quantity OneTM software. The software was used to determine the relative amount of each band in the sample row. The protein subunit bands of the glycine percentage and the β-conglycinin percentage are reported as a relative percentage of the total protein in the row. The α5-glycine subunit was not quantified and was not included in the total acid glycine value. Sample identities and weights are tracked using the LIMSTM Master.

Low odor producing soybean compositions with high β-conglycinin content are illustrated with some strains that lacked the low odor content (Table 22). Duplicate odor analyzes are indicated in the first row of the table. Several lines lacking glycine are indicated and produced compositions with less than 20 µg / g of hexane plus 2,4-decadienal plus hexanol per gram of ground seeds after oxidation under mild aqueous conditions. The compositions produced will be further evaluated for other characteristics (such as yield, free amino acids, color and fatty acid spectrum).

Table 22: Replenishment of Odor Production and Protein Subunits of High Beta-Conglycinin Soya Bean Compositions Deposit Information A deposit from Monsanto Technology LLC, soybean 0119149, described above and set forth in the claims, has been made. under the Budapest Treaty with the American Type Culture Collection (ATCC), 10801 University Boulevard, Manassas, VA 20110, USA. Deposit 0119149 deposited has the designations "A-411 and" 03JBK8-25 ", which are used in this descriptive report and examples and tables of work. ATCC accession number for deposit is PTA-6197 and date of deposit September 10, 2004. The deposit will be held at the depositary for a period of 30 years, or 5 years after the last application, or during the effective validity of the patent, whichever is longer, and will be replaced as necessary during that period. period.

All compositions and methods described and claimed herein may be produced and performed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it should be apparent to those skilled in the art that variations may be applied to the compositions and methods and to the steps or sequence of steps described herein, without departing from the concept of spirit and scope of the invention. More specifically, it should be apparent that certain agents, which are chemically and physiologically related, may substitute for the agents described herein, while achieving the same or similar results. All of these similar substitutions and modifications, apparent to those skilled in those techniques, are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

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CLAIM

Claims (1)

1. A method for analyzing the odor-producing properties of a soybean, which comprises determining the level of at least one compound selected from the group consisting of 2,4-decadienal, hexanol, hexanal and 1-octene. 3-ol, wherein determining the level of at least one compound comprises incubating a mixture of about 1 part soybean seed meal and about 4 parts water over a period of 1 to 40 minutes and quantifying the quantities of 2,4-decadienal, hexanol, hexanal and 1-octen-3-ol, or combinations thereof, using deuterated hexanal, hexanol and decadienal standards, in which soybean seed meal is manufactured from of peeled soybeans.