(21) Appl. No.: 11/577,241

Transcription

1 US A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2008/ A1 Deak et al. (43) Pub. Date: Apr. 24, 2008 (54) NOVEL VEGETABLE PROTEIN FRACTIONIZATION PROCESS AND COMPOSITIONS III (75) Inventors: Nicolas Alejo Deak, Ames, IA (US); Lawrence A. Johnson, Ames, IA (US) Correspondence Address: MCKEE, VOORHEES & SEASE, P.L.C. 801 GRAND AVENUE SUITE 3200 DES MOINES, IA (US) (73) Assignee: IOWA STATE UNIVERSITY RESEARCH FOUNDATION, INC., Ames, IA (US) (21) Appl. No.: 11/577,241 (22) PCT Filed: Oct. 20, 2005 (86) PCT No.: 371(c)(1), (2), (4) Date: PCT/US05/37943 Apr. 13, 2007 Related US. Application Data (60) Provisional application No. 60/620,922,?led on Oct. 21, Publication Classi?cation (51) Int. Cl. A23L 1/20 ( ) C07K 1/36 ( ) C07K 14/415 ( ) A23L 2/38 ( ) (52) U.s.C /598;530/418;530/378 (57) ABSTRACT According to the invention a novel vegetable protein frac tionation procedure is disclosed Which includes a straight forward process to obtain 7S-rich ([3-conglycinin-rich) and lls-rich (glycinin-rich) isolated protein fractions With unique functional and nutritional properties desired by the food industry. The process is much simpli?ed compared to the art and avoids multiple steps in the usual fractionation of soy protein and uses very small amounts of salts avoiding the necessity of excessive Washing and desalting steps. The process yields high amounts of protein fractions With high iso?avone contents and improved functional properties. Defatted Soy Flour 2N NaOH > v EXTRACT (1511, 1 111, 25 O0, ) CENTRIFUGE (14,000Xg, 3 0 min) 1 Supernatant (protein extract) < NaHSO; <- CaC12 * 2N HCl V PRECIPITATE (pl-i 6.4, optional store at 4 C overnight) v Spent Flour CENTRIFUGE (9,000Xg, 30 min) I Glycinin Curd NEUTRALIZE DRY Dried Glycinin-rich Fraction

2 Patent Application Publication Apr. 24, 2008 Sheet 1 0f 22 US 2008/ A1 2N NaOI-I > Defatted Soy Flour V EXTRACT Supernatant (15:1, 1 hr, 25 C, ph 8.5) (protein extract) < NaHSO3 < - CaC12 <' 2N HCl V CENTRIFUGE (14,000xg, 30 min) PRECIPITATE (ph 6.4, optional store at 4 C overnight) Spent Flour V CENTRIFUGE (9,000Xg, 30 min) i Glyeinin Curd NEUTRALIZE DRY Dried Glycinin-rich Fraction Fig. 7A

3 Patent Application Publication Apr. 24, 2008 Sheet 2 of 22 US 2008/ A1 Supernatant (protein extract) ACIDIFY (ph 4.8), CENTRIFUGE (9,000xg, 30 min) i B -cong1ycinin Curd > Whey NEUTRALIZE DRY Dried B-conglycinin rich Fraction Fig.

4 I _. _- l Patent Application Publication Apr. 24, 2008 Sheet 3 of 22 US 2008/ A1 Defatted Soy Flour 2N NaOH > i EXTRACT (15:1, 1 hr, 20 C, ph 85, remove solids by stainless-steel screen ~50 mesh) v CENTRIFUGE (14,000xg, I Supernatant (protein 15 C ref, 30 min) extract) 2N HCl - > 10 mm NaHSO; > Spent Flakes, PRECIPITATE (ph 6.4, l FREEZE DRY Dried Spent Flakes store at 4 C, l2-l 6 hr) v CENTRIFUGE (7,500xg, I Supernatant ( protein 20 min, 4 C) extract) Glyeinin Curd i l NEUTRALIZE : l NaCl 0425M > PRECIPITATE (ph 5.0, stir 1 hr) I , CENTRIFUGE FREEZE-DRY : (14,000xg, 30 min, 4 C) : (72 hr). u "l v l Dried Glycinin Intermediate curd l NEUTRALIZE : I" " " " " l - ' I : FREEZE- I : DRY l...i... l Dried Intermediate. Protein Fraction Optional step Fig. 2A

5 Patent Application Publication Apr. 24, 2008 Sheet 4 of 22 US 2008/ A1 Supernant (protein extract) H2O 2:1- > ACIDIFY (ph 4.8), QEN'ERIFUGE _> (7,500xg, 20 mm, 4 C) l B-Conglycinin curd Whey FREEZE-DRY (72 hr) L ' Dried l NEUTRALIZE : Whey l _ l _ i FREEZE-DRY i i (72 hr) Dried B-Conglycinin Fig. 2B

6 Patent Application Publication Apr. 24, 2008 Sheet 5 0f 22 Defatted Soy Flour l" _' ZNNaOH EXTRACT (10:1, 60 C, 30 min, ph 8.5) US 2008/ A1 CENTRIFUGE (14,000xg, SCREEN 15 C ref, 30 min) = (80 mesh) * Supernatant l 2N HCI Spent Flakes PRECIPITATE (2 min,, 20 C, mixing, ph 4.5) i l FREEZE-DRY (48 hr) 1 i l # Dry Spent Flakes Liquid CENTRIFUGE Whey REFRIGERATE (4 C, 1 hr) (14,000xg, 30 min, 4 C) l Isolate Curd l < H20 (211) 7 < 2N NaOH Dry Whey NEUTRALIZE FREEZE-DRY (48 hi) Fig. 3 Isolate

7 Patent Application Publication Apr. 24, 2008 Sheet 6 0f 22 US 2008/ A1 UGE SUCROSE - > UDP-GLU - > UDP-GAL SUCROSE FRUCTOSE GALACTINOL RS + GLUCOSE GLC-6P '_> MYO-INOSITOL V RAFFINOSE MI-lPS GALACTINOL ss V PHYTIC ACID STACHYOSE Fig. 4

8 Patent Application Publication Apr. 24, 2008 Sheet 7 of 22 US 2008/ A1 Reducing agent Alkali Defatted soy flour EXTRACT (ph '7 to 11) l CENTRIFUGE _ I Supernatant (protein extract) l < Divalent cationic salt/edta < - Acid Spent?our " PRECIPITATE (ph 5 to 7) i l Supernatant protein extract) CENTRIFUGE > ([i-conglycinin-rich Precipitate (Glycinin-rich curd) Fig. 5

9 Patent Application Publication Apr. 24, 2008 Sheet 8 of 22 US 2008/ A1 Defatted soy?our Alkali > v EXTRACT (ph 7 to 11) Supernatant w (pr- 611mm <_ Reducing agent < - Divalent cationic salt/edta _' Acid Spent flour PRECIPITATE (ph 5 to 7) Precipitate (Glycinin-rich curd) Water<"---1: MEMBRANE FILTER i Additives._>. Additives , HOMOGENIZE Glycinin-rich soy protein isolate Fig. 6A

10 Patent Application Publication Apr. 24, 2008 Sheet 9 0f 22 US 2008/ A1 Supernatant (B-conglycinin-rich protein extract) < - Acid v PRECIPITATE (ph 4 to 6) " Supernatant CENTRIFUGE i [i-conglycinin-rich curd l < - -- Alkali NEUTRALIZE (Lipoxygenase-rich, trypsin-inhibito r-rich protein extra ct) l r _ -.1 l : MEMBRANE FILTER - ~> Water L Additives JET-COOK v HOMOGENIZE Additives v DRY i [i-conglycinin-rich soy protein isolate Fig. 6B

11 Patent Application Publication Apr. 24, 2008 Sheet 10 of 22 US 2008/ A1 Defatted soy flour Alkali >" EXTRACT (pi-i 7 to 11) Supernatant CENTRIFUGE > (protein extract) 4 Reducing agent i ' Spent flour PRECIPITATE (ph 5 to 7) < Divalent cationic sait/edta v CENTRIFUGE Acid Precipitate,.l (Glycinin-rich curd) E Optional step 1 Alkall ->l : > : L ; NEUTRALIZE Water H: MEMBRANE FILTER 5 Protease, alkali, or acid > HYDROLYZE I v Additives > t JET-COOK Additives "V " HOMOGENIZE DRY Glycinin-rich soy protein hydrolyzate Fig. 7A

12 Patent Application Publication Apr. 24, 2008 Sheet 11 of 22 US 2008/ A1 Supernatant ([i-conglycinin-rich protein extract) i I _ \ E HYDROLYZE Protease, alkali, or acid V PRECIPITATE (ph 4 to 6) v CENTRIFUGE l B-conglycinin-rich curd l <- - Alkali NEUTRALIZE Supernatant (Lipoxygenase-rich, trypsin-inliibitor- rich protein extract) l, _. l 1 : MEMBRANE FILTER : ' ' ' ' ' > Water + HYDROLYZE < - Protease, alkali, or acid v JET-COOK Additives < Additives HOMOGENIZE DRY l?-conglycinin-rich soy protein hydrolyzate Fig. 7B

13 Patent Application Publication Apr. 24, 2008 Sheet 12 0f 22 US 2008/ A1 Defatted soy?our Alkali - > v EXTRACT (ph 7 to 11) CENTRIFUGE Spent flour Supernatant (protein extract) Reducing agent < Divalent cationic salt/edta _ _ - - Acid PRECIPITATE (ph 5 to 7) v CENTRIFUGE E Optional step E l i ' ' ' ' > i Precipitate """""""""" " (Glycinin-rich curd) Additives _> JET-COOK Add1t1ves > " HOMOGENIZE V DRY Isoelectric Glycinin-rich soy protein isolate Fig. 8A

14 Patent Application Publication Apr. 24, 2008 Sheet 13 of 22 US 2008/ A1 Supernatant (B-conglycinin-rich protein extract) 4 ' - Acid v PRECIPITATE (ph 4 t0 6) " Supernatant CENTRIFUGE i (Lipoxygenase-rich, trypsin-inhibito r-l ich protein extract) B-conglycinin-rich curd l 4 -- Additives JET-COOK v HOMOGENIZE Additives DRY l Isoelectric B-conglycinin-rich soy protein isolate Fig. 8B

15 Patent Application Publication Apr. 24, 2008 Sheet 14 of 22 US 2008/ A1 Defatted soy?our Alkali >" EXTRACT (ph 7 to l l) CENTRIFUGE i Supernatant > (protein extract) < Reducing agent Spent flour PRECIPITATE (ph 5 to 7) < Divalent cationic salt/edta. Acid v CENTRIFUGE > Precipitate,.l (Glycinin-rich curd) i Optional step : Alkali > i > i ' _ -1 NEUTRALIZE Water MEMBRANE FILTER 5 i. _ Protease, alkali, or acid > HYDROLYZE V Additives > v JET-COOK Additives > v HOMOGENIZE v DRY Glycinin-rich soy protein hydrolyzate Fig. 9A

16 Patent Application Publication Apr. 24, 2008 Sheet 15 of 22 US 2008/ A1 Supernatant (B-conglycinin-ri ch protein extract) l I E HYDROLYZE Protease, alkali, or acid _ r MEMBRANE Filtrate FILTER I (Salts and sugars) l Retentate B-conglycinin-rich extract E MEMBRANE FILTER {"--> Water I... -_ Additives * 4 JET-COOK 4 Additives HOMOGENIZE v DRY l Membrane filtered B-conglycinin-rich soy protein isolate/hydrolyzate Fig. 9B

17 Patent Application Publication Apr. 24, 2008 Sheet 16 of 22 US 2008/ A1 Defatted high-sucrose Alkali >l EXTRACT (ph 7 to 11) Spent flour Supernatant (protein extract) <._ Reducing agent < Divalent cationic salt/edta 4- Acid PRECIPITATE (ph 7 to 5) Supernatant ([i-conglycinin-rich protein extract) - Additives Precipitate (GlycinJ'n-rich curd) HOMOGENIZE 4 Additives r > ' Additives Additives : l\/iembrane FILTER :?-conglycinin-rich high-sucrose soy protein product Glycinin-rich soy protein isolate Fig. 70

18 Patent Application Publication Apr. 24, 2008 Sheet 17 of 22 US 2008/ A1 Partially defatted or full-fat soy flour Alkali ---> i EXTRACT (ph 7 to 11) CENTRIFUGE > Supernatant l Spent flour Supernatant (protein extract) PRECIPITATE (ph 5 to 7) - Reducing agent < Divalent cationic salt/edta < Acid i CENTRIFUGE - > Supernatant (B-conglycinin-rich ( cream fraction ) JET'COOK HOMOGENIZE I i protein extract) DRY Precipitate (Glycinin-rich curd) High-oil/High-protein product Fig. 11

19 Patent Application Publication Apr. 24, 2008 Sheet 18 of 22 US 2008/ A1 Alkali or acid Defatted soy flour >l EXTRACT (ph 6 to 10) Reducing agent -- > Divalent cationic salt/edta > Acid -> V PRECIPITATE (ph 5 to 7) ' Supernatant CENTRIFUGE - > (IS-conglycinin-rich l protein extract) Precipitate (Glycinin-rich?ber-rich?our/concentrate) Fig. 12

20 Patent Application Publication Apr. 24, 2008 Sheet 19 of 22 US 2008/ A1 Alkali or acid Partially defatted or full-fat soy?our * l EXTRACT (ph 6 to 10) Reducing agent > Divalent cationic salt/edta - > Acid > v PREClPITATE (ph 5 to 7) i CENTRIFUGE > Supernatant > Supernatant ( cream fraction ) l Precipitate (Glycinin-rich?ber-rich flour/concentrate) (Ii-conglycinin-rich protein extract) JET-COOK v HOMOGENIZE DRY High-oil/High-protcin Product Fig. 13

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