t Dried Glyeinin-rich Fraction

Transcription

1 (19) United States US A1 (12) Patent Application Publication (10) Pub. No.: US 2012/ A1 Deak et al. (43) Pub. Date: Apr. 19, 2012 (54) NOVEL VEGETABLE PROTEIN (60) Provisional application No. 60/620,922,?led on Oct. FRACTIONIZATION PROCESS AND 21, COMPOSITIONS Publication Classi?cation (75) Inventors: Nicolas Alejo Deak, Ames, IA (51) Int_ CL (liaémwrence A. Johnson, Ames, C07K 2/00 (200601) ( ) (52) US. Cl /378 RESEARCH FOUNDATION, INC, Ames, IA (Us) According to the invention a novel vegetable protein fraction ation procedure is disclosed Which includes a straightforward (21) App1_ NO; 13/315,437 process to obtain [3-conglycinin-rich and glycinin-rich iso lated protein fractions With unique functional and nutritional (22) Filed, Dec_ 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 Related U's' Apphcatlon Data amounts of salts avoiding the necessity of excessive Washing (62) Division of application No, 11 /577,241,?led on Apr, and desalting steps. The process yields high amounts of pro 13, 2007,?led as application No. PCT/US05/37943 on tein fractions With high iso?avone contents and improved Oct. 20, functional properties. Def-atted Soy Flour Y EXTRACT (15.1, 1 hr, 25 00, ph 8.5) Supernatant (protein extract) Supernatant (protein extract) CENTRIFUGE (1 4,000, g, 30 min) PRECTPITATE (pl-i 6.4, optlonal store at 4 C overnight) i " CENTRIFUGE Spent Flour (9j000xg7 30 min) i Glycinin ACIDIFY (ph 4.8), CENTRIFUGE (9,000xg, 30 min) v B-cnnglycinin Curd 4r Whey N EUTRALIZE DRY t Dried Glyeinin-rich Fraction NEUTRALIZE DRY V Dried B-conglycinin rich Fraction

2 Patent Application Publication Apr. 19, 2012 Sheet 1 0f 22 US 2012/ A1 2N NaOH > Defatted Soy Flour V EXTRACT Supernatant (15:1, 1 hr, 25 C, ph 8.5) (protein extract) < - NaHSO3 < - CaClZ < 2N HCl 7 CENTRIFUGE (14,000xg, 30 min) PRECIPITATE (ph 6.4, optional store at 4 C overnight) Spent Flour V CENTRIFUGE (9,0007%, 30 min) i Glycinin Curd NEUTRALIZE DRY Dried Glycinin-rich Fraction Fig. 7A

3 Patent Application Publication Apr. 19, 2012 Sheet 2 0f 22 US 2012/ A1 Supernatant (protein extract) ACIDIFY (ph 4.8), CENTRIFUGE (9,000xg, 30 min) i B-conglycinin Curd > Whey NEUTRALIZE DRY Dried B-conglycinin rich Fraction Fig. 1B

4 Patent Application Publication Apr. 19, 2012 Sheet 3 of 22 US 2012/ A1 Defatted Soy Flour 2N NaOH > i EXTRACT (15:1, 1 hr, 20 C: ph 8.5, remove solids by stainless-steel screen ~50 mesh) v CFNTRIFI IGE (14,000xg, I Supernatant (protein 15 C ref, 30 min) extract) 2N HCl m} 10 mlvi Nal-lSO; > Spent Flakes i FREEZE DRY Dried Spent Flakes I PRECIPITATE (ph 6.4, store at 4 C, hr) v CENTRIFUGE (7,500xg, I Supernatant (protein 20 min, 4 C) extract) NaCl 025M > Glycinin Curd PRECIPITATE (ph 50, stir 1 hr) - _. I NEUTRALIZE : l I I l CENTRIFUGE. FREEZE-DRY ; (14,000xg, 30 min, 4 C) { (72 hr) 1 i l Dried Glycinin Intermediate curd v NEUTRALIZE -- Optional step Dried Intermediate Protein Fraction Fig. 2A

5 Patent Application Publication Apr. 19, 2012 Sheet 4 0f 22 US 2012/ A1 Sup Brnant (protein extract) {-120 2:l > ACIDIFY (ph 4.8), CENTRIFUGE _> (7,500xg, 20 min, 4 C) l B-Congiycinin curd I Whey l FREEZE-DRY (72 hr) Dried i NEUTRALIZE i Whey _ - - _ i i """" ' 'l' """ ' I I FREEZE-DRY : I (72 hr) : Dried IS-Conglycinin Fig. 2B

6 Patent Application Publication Apr. 19, 2012 Sheet 5 0f 22 US 2012/ A1 Defatted Soy Flour EXTRACT (10:1, 60 C, 30 min, ph 8.5) V CENTRIFUGE (14,000xg, SCREEN 15 C ref., 30 min) > (80 mesh) w Supernatant i 4 2N HCl Spent Flakes PRECIPITATE (2 min,, 20 C, mixing, ph 4.5) i i REFRIGERATE (40c, 1 hr) V Dry Spent Flakes Liquid CENTMFUGE Whey (14,000xg, 30 min, 4 C) I F REEZE-DRY (48 hr) 2 Isolate Curd 5 <~ - H20 (2; 1) 7 < 2N NaOH Dry W h ey l NEUTRALIZE Fig. 3 FREEZE-DRY (48 hi) i Isolate

7 Patent Application Publication Apr. 19, 2012 Sheet 6 0f 22 US 2012/ 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. 19, 2012 Sheet 7 of 22 US 2012/ A1 Defatted soy flour Reducing agent ---~ ~> Alkali w; EXTRACT (ph '7 to 11) l CENTRTFUGE > l Spent?our V Supernatant (protein extract) < Divalent cationic salt/edta <-_ Acid PRECIPlTATE (ph 5 to 7) i l Supernatant protein extract) CENTRIFUGE > (?-conglycinin-rich Precipitate (Glycinin-rich curd) Fig. 5

9 Patent Application Publication Apr. 19, 2012 Sheet 8 0f 22 US 2012/ A1 Defatted soy flour Alkali > v EXTRACT (ph 7 to 11) Supernatant w emblem» < Reducing agent Spent flour 4 - Divalent cationic salt/edta I Acid PRECIPITATE (ph 5 to 7) Optional step - -- Precipitate (Glycinin-rich curd) Water < MEMBRANE r FILTER Additives - ~ > ' Additives e _> HOMOGENIZE Glycinin-rich soy protein isolate Fig. 6A

10 Patent Application Publication Apr. 19, 2012 Sheet 9 of 22 US 2012/ A1 Supernatant (B-conglycinin-rich protein extract) 4" -"~ - Acid v PRECIPITATE (ph 4 to 6) y CENTRIFUGE Y [3-conglycinin-rich curd i < Alkali NEUTRALIZE (L. Supernatant. h 1p0xygenase-r1c, trypsin-inhibito r-rich protein extra ct) i MEMBRANE FILTER > Water T"'IIIIII Additives JET-COOK v HOMO GENIZE 4 " ' Additives v DRY Y [i-conglycinin-rich soy protein isolate Fig. 6B

11 Patent Application Publication Apr. 19, 2012 Sheet 10 0f 22 US 2012/ A1 Defatted soy?our Alkali EXTRACT (pl-i 7 to 11) Supernatant 4... Reducing agent Spent flour PRECIPITATE (ph 5 to 7) < Divalent cationic salt/edta ' Acid Precipitate, _ (Glycinin-rich curd) E Optional step i Alkali >l * i w Protease, alkali, or acid Water "4: MEMBRANE FILTER i * HYDROLYZE Glycinin-rich soy protein hydrolyzate Fig. 7A

12 Patent Application Publication Apr. 19, 2012 Sheet 11 of 22 US 2012/ A1 Supernatant ( 3-cong1ycinin~rich protein extract) E HYDROLYZE E1" Protease, alkali, or acid uuuuuu 4 -~ Acid PRECIPITATE (ph 4 to 6) v CENTRIFUGE l B-conglycinin-rich curd i < ~ Alkali NEUTRALIZE Supernatant (Lipoxygenase-rich, trypsin-inhibitor- rich protein extract) i MEMBRANE FILTER E > Water HYDROLYZE <~ Protease, alkali, or acid Additives JET-COOK HOMOGENIZE 4 ~~ Additives DRY i [3 c0nglycinin~rich soy protein hydrolyzate Fig. 7B

13 Patent Application Publication Apr. 19, 2012 Sheet 12 of 22 US 2012/ A1 Defatted soy flour Alkali - > V EXTRACT (ph 7 to 11) Supernatant M (protein extract) 4 Reducing agent < Divalent cationic salt/edta 4_' Acid Spent flour 7 PRECIPITATE (ph 5 to 7) Optional step ' > Precipitate (Glycinin-rich curd) Additives _> Additives > HOMOGENIZE Isoelectric Glycinin-rich soy protein isolate Fig. 8A

14 Patent Application Publication Apr. 19, 2012 Sheet 13 of 22 US 2012/ A1 Supernatant ([3-conglycinin-rich protein extract) Acid PRECIPITATE (ph 4 to 6) ' CENTRIFUGE Supernatant (Lipoxygenase-rich, trypsiminhibito rich protein extract) V B-conglyeinin-rich curd l Additives JET-COOK v Additives HOMOGENIZE DRY Isoelectric?-conglycinin-rich soy protein isolate Fig. 8B

15 Patent Application Publication Apr. 19, 2012 Sheet 14 of 22 US 2012/ A1 Defatted soy flour Alkali EXTRACT (ph 7 to l l) Supernatant (protein extract) 4._..._. Reducing agent 4 Divalent cationic salt/edta w Acid Spent flour PRECIPITATE (ph 5 to 7) Precipitate,.I (Glycinin-rich curd) i Optional step : Alk?ll * a > ; Water MEMBRANE FILTER i,. I V Protease, alkali, or acid > HYDROLYZE I Additives > Additives ~> v JET-COOK Glycinin-rich soy protein hydrolyzate Fig. 9A

16 Patent Application Publication Apr. 19, 2012 Sheet 15 0f 22 US 2012/ A1 Supernatant (B-conglycinin-ri ch protein extract) l i HYDROLYZE i4 t - - Protease, alkali, or acid _ I. V IVLEh/IBRANE Filtrate FILTER ' (Salts and sugars) Retentate B-conglycinin-rich extract l r w H H I i MEMBRANE FILTER I----> Water """"" "PI-1:111 - Additives i JET-COOK 4. w. _. Additives Membrane filtered B-conglycinin-rich soy protein isolate/hydrolyzate Fig. 9B

17 Patent Application Publication Apr. 19, 2012 Sheet 16 0f 22 US 2012/ A1 Defattcd high-sucrose soy?u ur Alkali EXTRACT (ph 7 to 1 1) CENTRlF U GE Spent?uur Supernatant (protein extract) 4. Reducing agent -4 Divalent cationic salt/edta 4"' Acid PRECIPITATE (ph 7 to 5) Supernatant w protein extract) 3 Optional step i i """"" ~ ' Addltlves : > 5 ' ' Precipitate 4- Additives (Glycim'n-rich curd) HOMOGENIZE Additives?-conglycinin-rich high-sucrose so rotein roduct Additives y P p Glycinin-rich soy protein isolate Fig. 70

18 Patent Application Publication Apr. 19, 2012 Sheet 17 of 22 US 2012/ A1 Partially defatted or full-fat soy flour Alkali ~ > l EXTRACT (ph 7 to 11) CENTRIFUGE V A Supernatant i Spent flour bupematallt (protein extract) PRECIPITATE (ph 5 to 7) < -- Reducing agent < Divalent cationic salt/edta < - Acid l CENTRIFUGE -> Supernatant ( 3 c0nglycinin-rich ( cream fraction ) JET'COOK HOMOGENIZE l protein extract) DRY Precipitate (Glycinin-rich curd) High-oil/High-protein product Fig. 1 7

19 Patent Application Publication Apr. 19, 2012 Sheet 18 of 22 US 2012/ A1 Alkali or acid Defatted soy flour > i EXTRACT (ph 6 to 10) Reducing agent -- > Divalent cationic salt/edta > Acid -> " PRECIPITATE (ph 5 1:0 7) 7 Supernatant CENTRIFUGE - > (B-conglycinin-rich l protein extract) Precipitate (Glycinin-rich fiber- rich?our/concentrate) Fig. 12

20 Patent Application Publication Apr. 19, 2012 Sheet 19 of 22 US 2012/ A1 Alkali or acid Partially defatted or full-fat soy flour >l Reducing agent - > Divalent cationic salt/edta ~ > Acid - > EXTRACT (ph 6 to 10) PRECIPITATE (ph 5 to 7) CENTRIFUGE > Supernatant > Supernatant ( cream fraction ) ([3~conglycinin-rich protein extract). JET-COOK Precipltate (Glycinin-rich?ber-rich Y?our/ concentrate) HOMOGENIZE High-oil/High-protein Product

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