REF. No.	SAMPLE	EXTRACTION METHOD
48	soybean	-Soy Isolate: extract defatted flour(1:9) in H2O, at pH 9.0 w/ 1N NaOH, fed to centrifugal desludger. Resuspend residues w/ H2O (1:10), done 3 times. Collect supernatant adjust to pH 7.0 w/ 6N HCl, put into ultrafilter w/ hollow unit fiber.
55	soybean	-extract 3 g flour in 20 mL H2O, refrigerate overnight , add 10 mL 0.1M phosphate buffer (pH 7.6), 1.2M NaCl, 3mM DTT. Stir in RT/ 60 min, centrifuge at 10000xg/ 30 min. Add acetone to filtrate 1.5 fold at -20 C dropwise, centrifuge at 10000xg/ 30 min in cold, 2 times. Dried precipitate in vacuum oven.
78	soybean	-make bean slurry (1:10) w/ H2O, boil for 15 min, filtered w/ 4-layer cheese cloth. Cooled to 70 C, add 400 mL 10% CaSO4.2H2O. Set for 10 min, press in cheese cloth.
33	soybean	-extract defatted flour (1:10) in H2O, centrifuge at 12000xg /10 min. Bring filtrate to pH 4.5, recentrifuge at 10000xg/ 10 min. Adjust precipitate to pH 7.0 and freeze dried.
58	soybean	-extract defatted flour (1:20) in H2O at RT, centrifuge at 15000xg/ 25 min at 5 C, collect the water soluble protein. Add (1:20) 0.5M NaCl to residue, centrifuge at RT, done 4 times, as NaCl soluble protein.
82	soybean	-extract defatted flour (1:14) in H2O, adjust to pH 8.5 with 6N NaOH, 30 min. centrifuge at 2000 rpm/ 15 min. Adjust filtrate to pH 4.5 with 6N HCl, 30 min., centrifuge. suspended precipitate in H2O, pH 7.0, free dried. Or: to collect 11S, add 0.03M Tris buffer to ppt. (pH 6.4) at 1 C, centrifuge. Bring filtrate to pH 4.8, 7S will be ppted.
19	soybean	-make slurry of defatted flour in H2O, pH 8.0, centrifuge. Bring filtrate to pH 4.5 w/ HCl, wash precipitate w/ H2O, adjust to pH 7.0 w/ NaOH, spray dried.
66	soybean	-Protein concentrate: extract defatted flour (1:10) in H2O at 40 C/ 30 min. Add 3N HCl to pH 4.5 and store overnight at 7 C. Remove whey, repeat 3 times, freeze dried. Or: add 20% NaOH to pH 7.0, freeze dried
		-Water wash concentrate: extract defatted flour (55g:65mL) in H2O, spread paste in st.steel pan, autoclave at 121 C/ 5 min. Grind and disperse in H2O (1:10), heat suspension to 82 C/ 15 min. Remove whey, repeat 2 times. Centrifuge and Freeze dried.
		-Tofu Analog: extract defatted flour (1:10) in H2O, heat to 90 C/5 min, cooled to 70 C. Add CaSO4 (2.25 g + 18 mL H2O for 420 mL slurry), mix rapidly. Incubate at 70 C/ 10 min. Broke curd, centrifuge at 100xg/ 5 min. Resuspend ppt., repeat 2 times. Freeze dried ppt.
		-Protein Isolate: extract whole flour (1:10) in H2O, bring to pH 9.0 w/ 20% NaOH, stirat 35 C/ 45 min. Passed through centrifugal desludger. Adjust extract to pH 4.5 w/ 3N HCl, stored overnight at 7 C. Remove whey, done 3 times, freeze dried the ppt. , or add 20% NaOH to pH 7.0, freeze dried.
79	soybean	-extract soy flake (1:10) in H2O, sonicated in chilled for 8 min., centrifuge at 10000xg/ 15 min. Take supernatant, lyophilized.
65	yellow pea	-extract flour (1:5) in H2O, add 1N NaOH to pH 9.0, stir 20 min., centrifuge 1000xg/20 min. Done 2 times. Bring combined filtrate to pH 4.5, centrifuge and redisperse ppt. w/ 100 mL H2O at pH 9.0, reprecipitate, and dried. Or: add 1N NaOH to ph 6.5-7.0, freeze dried.
27	yellow pea	-extract flour (1:10) in H2O, add NaOH to pH 8.5 at 25 C/ 30 min., done 2 times. Collect extract, add HCl to pH 4.5, centrifuge 1570xg at 10 C/ 10 min.
25	legumes (yellow pea, lentils, faba beans)	-extract flour (1:10) in H2O, to pH 8.5 w/ 1N NaOH at 40 C /30 min. done 2 times. Centrifuge at 220xg at 10 C/ 10 min. Combined filtrate, add 1N HCl to pH 4.5, centrifuge at 1570xg at 10 C/ 10 min. Wash ppt. w/ H2O , add 1N NaOH to disperse ppt. to pH 7.5. Dialyzed in running tap water at 10 C/ 24 hr, lyophilized.
21	fieldpea	-extract flour (100 g/500 mL) in H2O, add 0.2% CaO to pH 8.8-9.0, centrifuge at 100xg/20 min. Heat to 95-100 C/ 20 min, cooled to 75-80 C for 2 sec. Coagulate w/ 0.54% CaCl2, HOAc, CaSO4.
75	chick pea	-extract flour (1:8 to 1:20) in H2O, at pH 2-12. Apply continuous centrifugation. Soluble solids were concentrated in hollow fiber ultrafiltration unit. Spray dried.
68	mung bean	-Protein Isolate: extract flour (1:15) in H2O, add 1N HCl or NaOH to pH 1-12. Shake in waterbath 25 C/ 20 min, centrifuge at 1000xg/ 20 min, at RT. Add 1N HCl to filtrate, pH 4.0, centrifuge. Add ppt. w/ 1N NaOH to pH 7.0.
50	winged bean	-Isoelectric protein isolate: extract defatted flour (1:20) in H2O at ph 10, 90 min/ 30 C, centrifuge at 10000 rpm /30 min. Precipitate at pH 4.0.
	-Neutralized protein isolate: adjust to pH 12 at 30 C/ 30 min, centrifuge. Adjust filtrate to pH 7.0. Dried.
69	adzuki bean	-Albumin fraction: extract flour (1:10) in H2O at 4 C/ 24 hr, centrifuge at 10000xg/ 30 min. Dialyzed filtrate in H2O at 4 C/ 24 hr, freeze dried
		-Globulin fraction: add 2% NaCl (1:10) to residues at 4C/ 24 hr, centrifuge 10000xg/ 30 min , dialyzed in H2O at 4 C/ 24 hr, freeze dried.
		-Protein Isolate: extract flour (1:10) in 0.01N NaOH, at pH 8.5/ 2 hr, centrifuge at 3000xg/ 2 min. Add 1N HCl to pH 4.0 to thefiltrate, freeze dried the ppt.
83	cotton seed	-12S fraction: extract defatted flour w / H2O, 2 times, remove non storage protein. Add Tris-HCl 0.5M, NaCL 0.5 M, and 0.2 g/L NaN3, adjust to ph 7.0. Dilute supernatant w/ H2O (1:10), centrifuge to ollect ppt. as 12S.
	-7S fraction; Chilled filtrate to 0 C, centrifuge at 0 C. Dissolve ppt in 0.5 M Tris-0.5M NaCl, adjust to pH 7.0, as 7S.
24	soybean	-extract defatted flakes (1:5 and 1:10) in H2O and in alkali, at pH 8.0 w/ 2N NaOH for 40-60 min. Centrifuge at 10000 rpm (16000rcf)/ 20 min. Add filtrate w/ 2N HCl to ph 4.5, centrifuge. Wash ppt. then freeze dried as Isoelectric protein isolate. Or: adjust ppt. to pH 8.0, freeze dried as Neutalized PI.
7	okra seed	-Protein concentrate: extract defatted flour (1:20) in H2O at pH 10 using 6N and 1 N NaOH at 25 C/ 60 min. centrifuge at 1293xg/ 10 min.adjust filtrate to pH 7.0, freeze dried .
		-Protein isolate: adjust supernatant to pH 4.5 w/ 6N and 1N HCl, centrifuge at 1293xg/20 min. Resuspend ppt. in H2O at pH 7.0, then freeze dried.
35	soybean	-extract defatted flour (1:10) in dilute NaOH, pH 12 at RT/ 1 hr, centrifuge at 2000 rpm (600xg)/ 20 min. Dialyzed against H2O for 100 hr at 4 C, change H2O after 60 hr. Add ppt. w/dilute HCl to pH 4.5, centrifuge at 2000 rpm/ 20 in. then freeze dried the ppt.
2	legumes (navy, pinto bean, and lentils)	-extract flour (1:10) in 0.02% NaOH at RT for 1 hr., centrifuge at 1500xg at RT/ 15 min., done 2 times. Adjust combined filtrate to pH 4.5 w/ HCl concentrate, centrifuge at 1500xg/ 15 min. Wash ppt w/ H2O, freeze dried.
16	navy bean	-extract flour (1:10) in 0.02N NaOH at RT/ 1 hr., centrifuge, done 2 times. Add HCl concentrate to filtrate, pH 4.5, centrifuge at 1500xg/ 15 min. Wash ppt. w/ H2O, freeze dried.
56	lupin seed	-extract flour (1:5) in 0.02N NaOH at 21 C/ 16 hr, centrifuge at 10000xg at 4 C/ 30 min, done 2 times. Dialyzed combine filtrate against H2O for 72 hr. changes H2O 6 times. Freeze dehydrated.
57	northtern bean	-extract flour (1:10) in 0.5% Na2CO3 at 4C/ 24 hr, centrifuge at 10000xg/ 30 min. Dialyzed filtrate against H2O for 48 hr, change H2O 6 times. Lyophilized.
85	cowpea	-extract flour (1:10) in phosphate buffer, pH 6.8, let stand for 20 hr in refrigerator (2 C). Blend in Waring blender at medium speed for 15 min, done 2 times. Filter with cheese cloth. Centrifuge combined filtrate at 1000 rpm/ 1 min to ppt. starch. Add filtrate w/ 5% HCl to pH 4.5, let stand in refrigerator for 60 min. Centrifuge and wash w/ H2O. Vacuum dried the ppt. at 50-60 C/ 5-7 hr.
74	rapeseed	-extract flour (1:18) in 1% sodiun hexametaphosphate solution, pH 7.0. Stir in RT/ 30 min., adjust to pH 7.0 w/ 1-6N HCl or NaOH. Vacuum filtered to recover extract, put to ultrafiltration, ion exchange, then dried.
6	soybean	-7S fraction: extract defatted flour (1:20) in dilute Tris, centrifuge and adjust filtrate to pH 7.8. Freeze dried.
24	soybean	-11S fraction: add 0.03M Tris-HCl to defatted flour, at pH 8.0. Centrifuge at RT, bring filtrate to pH 6.4, recentrifuge at 2-5 C, collect ppt.
		-2S fraction: bring the filtrate to pH 7.6 w/ 0.03M Tris-HCl, extract is 2S.
		-7S fraction: add ppt. w/ Tris-HCl to pH 6.2, centrifuge. Put filtrate to Sepharose 6B to purify 7S.
67	soybean	-extract flour (1:10) in 60% ethanol, heat to 57 C/ 30 min. Remove supernatant, replace w/ 60% ethanol, repeat 2 times. Centrifuge the protein rich sludge, dried the ppt. to remove residual ethanol.

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Updated: Wednesday, June 20, 2007.