TEXAS AGRICULTURAL EXPERIMENT STATION R. D. LEWIS, Director College Station, Texas BULLETIN NO. 692 SEPTEMBER 1947 The Essential Amino Acid Content of Cottonseed, Peanut and Soybean Products CARL M. LYMAN, KENNETH KUIKEN and FRED HALE With the technical assistance of Shirley Dieterich, Marjory Bradford and Mary Trant AGRICULTURAL AND MECHANICAL COLLEGE OF TEXAS GIBB GILCHRIST, President V.» V_ , J10-1047-5M-L180 r5‘ I [Blank Page in Original Bulletin] Preface Amino acids are the chemical structural units from which various I» of proteins are built. The requirement of animals for protein in fact, a composite requirement for a number of different amino This report gives the content of 1o different‘ amino acids in 11s, flours and commercial protein preparations made from cotton- ‘_ , peanuts and soybeans. The following 1o amino acids: arginine, f 'dine, isoleucine, lcucine, lysine, methionine, phenylalanine, V, - nine, tryptophane and valine were selected for study on the basis their proven importance in nutrition. i- kind were quite uniform with respect to their amino acid position. _ eals and flours made from cottonseed, peanuts and soybeans are ,3: sources of amino acids. The protein of cottonseed and soybean F ucts was found to be definitely superior to the protein of peanut k ucts with respect to lysine, methionine and tryptophane. These f ino acids are of particular importance because feeds and feed ucts are likely to be deficient in one or more of them. v ith the exception of soybean protein and soybean feed, different 1- of commercial processing appeared to have little or no effect fthe amino acid composition of the products. en expressed on an equal protein content basis, meals of the " CONTENTS _ Page Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Materials and Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Description of Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. e5 Methods of Analysis .‘ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 6 Results and. Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . ." . . . . . . . . . . . . . . . . . . . . . . 11 Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 .._-W-.-~.-w,,...,.,-.-wpw a-vsnww.“ < . < < BULLETIN NO. 692 SEPTEMBER 1947 The Essential Amino Acid Content of Cottonseed, Peanut and Soybean Products CARL M. LYMAN, KENNETH KUIKEN and FRED HALE With the technical assistance of Shirley Dieterich, Marjory Bradford and Mary Trant The nutritional requirement of animals for protein is in reality a requirement for certain amino acids which are present in the pro- tein. Although there are over 2o recognized amino acids, only 1o are generally regarded as essential in animal nutrition. These IO amino acids are: arginine, histidine, lysine, valine, leucine, isoleu- cine, methionine, phenylalanine, threonine and tryptophane. Com- plete information on the amino acid content of feeds is needed as a guide in compounding rations for firm animals, if amino acid deficiencies are to be avoided. i p Data to be found in the literature on the amino acid composition of cottonseed, peanut and soybean products are rather scattered and limited. Most of the older chemical methods for the determination of amino acids are more applicable and more reliable when applied to purified proteins than when applied to foodstuffs containing carbohydrates and other substances besides proteins. Comprehen- sive studies on the amino acid composition of natural and manu- factured food products have been made practical by the develop- ment of microbiological assay methods during the last few years. In the present investigation microbiological procedures have been applied to the study of various products made from cottonseed, peanuts and soybeans. Materials and Methods Description of Samples With the exception of 2 samples of solvent extracted cottonseed meal all of t-he materials were commercial products. Only materials of first quality were included. Cottonseed meal samples graded _ prime, according to the rules of the National Cottonseed Products Association. The process used for the preparation of 2 samples of solvent extracted cottonseed meal (A&M process) has been described by Harris, Bishop, Lyman and Helpert ... . . . . . . . .........................WPOQ@OHQH~NQQ~AQMHO%QN@HQ>< . mm. . . m. . Wm . . . . . . . . . . . . . . . . . . . . . . Iwwwm W . 1N b-w m -§ - Q u u - u ¢ ¢ - u - ¢ u - ¢ I u n ¢ -.| u? I 5m Sq 2N s: 3N. S; $2 _. . m u \ . . . . . . . . . - . . ' . . . . . . . . . - - . . m1 . . . . . . . . . . . . . . . . . . . . . . . . . . .@H§0>@Oww éflwg S . . . . . . . . . . . . . . . . . . . . . . . . fiNQE mNL .. . . . . . . . . . . . . . . . . . fiflwg E . . . . . . . . . . . . . . . . . . . . . . . fiNQS m cwwnzow R m . . . . . ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .WHU5.@OMQ PmZMNQQ Mal“ Qwdhw>< E . N.‘ . . . @.m . . . . . ... . . . . . . . . . . ..->.-.UA:.NM~%Q.N.Q wmm 3H mm a; $3 m2. M; ._ @=_2 n .€ . . - . . . . . . . . - . - . . . R . . . . . . . ... . . . . . . . . . . . . . . . . . . . . . . . ..H5O7@ m 4 uszwmm w - - . - . . ~ . . . . .. . . . . . . . . . . . % I - . - - o - . . - - - - - - - - - - - . - . . - . - I mm v m. m mm o w R ll . m . . . . . . . . . . . . . . . . . . . . . . . . s mwa $4 gm. ma... 3a I34“ M32.“ ..x..%::flHHHHHHH.QQ==N~PAQIQ¢E m 3Q 2: mm; i: $2 2e $2 .. H. 4mm TQ=_=£E¢ 182 E NN . Q 5 . fi , we . N 2 . H S . 2 3.. . v K . 2 . . . . . . . . . . . é==2PEV 1&2 T . . - - - - - - . - - - . . - ~ . » 2, i: 3N Mm . N m“ . H $2 M: .m 3.3 svzxzn 120.656..» awqsfiea aszowv RQE mm . . . . . . . . . . . . . . . . . . . . N . a . m . . . . . . . AmMQOOMQ Juwwvdhuxw HG0>~Omv HNQS 0. $4. mm . N S a m” . H 3 . S g6 3 4a ..... . . Awmwvoma. 23 dwfiwwfixw 3020a .133 N 3Q mma 51w 3A 3W3 36 EYE . . . . . . . . . . . . . . . . . . . . Aofizmv 52m N wwwmcoupoO 1m 2~8~3 pcwvhma . pcwohwa fiEPSQ pnwokwa “smegma fiEBQQ m cmwpogn 29:5 smfioha Bafiam cmwpokm @383 w 25.8 E m5 ...: QUPU E ma» E 35.6 E ma» E Ema x Zv Ewpoi $255.5 @593 wfiwmummfl mficwwk< 3:50 muuswai naonmom 3E 35:5 ioownctoo u: “nuance ufimb was 051:2: £55m: A oSah ESSENTIAL AMINO ACID CONTENT OF THREE PRODUCTS 7 Methods of Analysis The amino acid analyses reported here were a.ll carried out by the use of microbiological procedures developed at the 'l‘exas Agri- cultural Experiment Station. Valine, leucine and isoleucine were determined by the use of Lactobacilltts arcz/rtnostts 17-5 as described i. by Kuiken, Norman, Lyman, Hale and Blotter Histidine and threonine were determined with Streptococcus faccalis R. Details of the procedures are given in a report by Lyman, Kuiken and Hale (3). Methionine was determined with Lcuconostoc mescntcroidcs P60 (see Lyman, Moseley, Butler, Wood and Hale Trypto- phane was determined with Streptococcus faccalis R (see Kuiken, Lyman and Hale Description of the methods for the deter- mination of the remaining amino acids will be given in a later publication. Results and Discussion There are several ways in which the amino acid composition of foodstuffs can be expressed. The data-presented in this report are’ given in 2 ways; first, as the percent of the individual amino acid in the sample, and second, as the percent of the amino acid in the protein. The first of these 2 Ways has the advantage that it gives a direct measure of the total amount of a given amino acid contained 1 in the sample. Since amino acids are the constituents from which proteins are made, it is to be expected that differences in the protein ; content of different samples of the same kind of foodstuff will be a reflected in corresponding differences in amino acid content. By i, dividing the percentage of a given amino acid in a sample by the I protein content of the sample and multiplying by I00 we obtain. the percentage of the amino acid contained in the. protein. It is this latter value which is the more useful in evaluating the nutritional 1 qualities of the protein in different kinds of foodstuffs. - It will be seen from the 3 tables that the content of the individual amino acids in thecottonseed meal samples decreased rather con- sistently with decreased protein content. The data also show that is the protein in the cottonseed products of different protein content, f and made by different commercial processes, all had practically the same‘ amino acid composition. It follows that the content of any 1 of these 1o amino acids in other samples of cottonseed meal can be {satisfactorily predicted on the basis of the average values given in i the ‘tables and the protein content of the sample. This is probably true for peanut meal and soybean meal as well, although a smaller “number of samples of these products were included in this investi- I gation. The data given in the tables indicate that this generaliza- 8 BULLETIN NO. 692, TEXAS AGRICULTURAL EXPERIMENT STATION .......... .......... ..............................WHOS.@CHQQNQQRAOMHOQPQNNHQ>< . mfim £3 Nwm mmm Sim mma 3% . 2m Ed mms 3mm mmm mma mm? . . . . . . ........Afis>_$I~@2 Nmm mma mm.» mm...“ mmm mma Nmmm . .. ....Am=@>_owv 182 Nmm gm s: 3pm flwm mma Nmmv ............é.==mimmmmv12.2 8m bma EL. mm...“ mmm S.“ $.13 . .. . .. . .......@==flm>mv_.$2 5am mma m3. mmm mmm 2mm 3mm .. .. ........AQ=_=2mmmmZ82 nwwnmom .......... .... .. . . ..- ... ...... .. . ........w#Ufi;uQHQPWZHNQQHQ%Q@NH@>< 5+ mm; mmm m“; mme mm; mmmm . . . .. . . . . ...........émi=.fi?mmimw2 3Q 3.4 iwm mmw S4. s: 5mm ........H....§_=m.~mmmmvQ82 mme 3N 3am Se 3+ mma 2.4m . .. . . . . . . . .....So_m _ iibmwm Hm.m HHAw .. .. mm.fi . .. .. .. . 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The explanation of this finding is as follows: Products of high protein a content, such as commercial soybean protein, are made from the oil seed meals by extraction of the protein with dilute salt or alkaline solution. The protein is then recovered from solution by precipita- tion. The residue left after the extraction of the protein is sold . for feed. The net result is a partial separation of the different ‘ kinds of protein contained in the original meal. It is, therefore, l logical to expect some differences between the amino acid composi- tion of the protein in the product of high protein content and in 1 the feed. The data given in the tables show that cottonseed, peanut and 5 soybean products are good sources of amino acids. Of the 1o amino acids studied in this investigation, lysine, methionine and trypto- * phane are perhaps the most important from the nutritional stand- point. Feeds and feed products are more likely to be deficient in > one or more of these 3 amino acids than in any of the other amino a acids studied. The amounts of these 3 amino acids contained in_ the protein of cottonseed and soybean products were significantly higher than the amounts found in peanut products. p The older chemical data on the amino acids contained in cotton- seed, peanut and soybean proteins have been summarized by Block 1 and Bolling Microbiological methods have been applied to the jrdetermination of tryptophane in some of the products studied in ‘l the present investigation, by Greene and Black (7), Stokes, Gunnes, Dwyer and Caswell (8), Wooley and Sebrell (9), Baumgarten, lMather and Stone (I0), and by Greenhut, Schweigert and Elve- {hjem (II). The value of 1.73 given in the last mentioned report, %for the tryptophane content of the protein of soybean meal, is in “excellent agreement with the results reported here. In all other pgcases the tryptophane values obtained in the present investigation are substantially higher than those to be found in the above men- tioned reports. It is unlikely that differences in thesamples studied ‘are responsible for these discrepancies. The more probable explana- Ition is to be found in the methods used for the hydrolysis of the smaterials. Preliminary tests in the present investigation likewise ligave lower results. The higher values reported here were obtained only after an extended study of the factors which influence the rfstability of tryptophane during the alkaline hydrolysis of proteins d foodstuffs. The details of these studies have been published Q.- ‘c’? 3 D" (D "1 (D /-\ (J1 \./ p‘ Values for the content of one or more of the other essential amino cids in some of the products studied here are to be found in the . i? $4. . . 1.1 .. 51.31 l. ......iu.Jiiaqli1iql-4su.s1iil§wij~iil= .x7|..._...1 .. l, i. ... . mwlfi 01000000- ---.-. - ¢ . - - - - .. -..-.-- --»_-.- 4.N4 444.44 N44.N NN4 N4.N N444 E4 444444 444mg Hmxunw................4466r.4 N 444.4 N44 N... .N NN.N . NNN 444.4. 4.4.4 N44 44 44N N4. . .........4.4N4.£6E&4444 44464244 o N444 444244 4N.N NN4 NN.4. NN.N N4.4 4444.44 $444. p . . . . . . . 44.446.44.84 482 n N444 NNH44 N444. N444 $4. NNHN 4.N4 9444 NN.N4. . . . . . . . . . . . . . . .44446>444w4 482 m N... 4 444. 44 4444 N N4. 4 N44 4. N4 N 4.N 4 NN 44 NN N4. . . . . . . . . . . 4644444444544 482 s E24 4.44.44 NN.4. 444.4 N24. 4444.N N4..4 4444.44 4.N.N4. .. . .4644E....444.4444 482 T 4444.4 N4244 NNN 4424 4.44.4. 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ESSENTIAL AMINO ACID CONTENT OF THREE PRODUCTS 11 following publications: Kuiken, et aL, (2), Stokes (8), Baum- A garten (Io), Greenhut, Schweigert and Elvehjem (12), Riesen, i; Schweigert and Elvehjem (13), Schweigert (I4), Horn, jones and Blum (I5). In some cases there is good agreement between the values given in these reports and those obtained in the present investigation. Marked discrepancies occur in the case of methionine. This problem has been extensively studied and evidence for the ; validity of the method used here has been reported The values for the valine, leucine and isoleucine content of cot- tonseed, peanut and soybean meal given in a previous report from the authors’ laboratory are lower than those given‘ here. In l this case the cause of the discrepancies was definitely found to be due to the inadequacy of the method used in the previous investi- T gation for the hydrolysis of the samples. Summary The content of IO different amino acids in I9 samples of cotton- seed, peanut and soybean products is given. The products include cottonseed flour, both hydraulic and solvent extracted cottonseed meals, peanut flour, peanut meal, both hydraulic and solvent ex- .tracted soybean meals, soybean protein and soybean feed. With respect to 3 amino acids which are of particular significance ; in nutrition, lysine, methionine and tryptophane, the proteins of ‘cottonseed and soybean meals were found to be definitely superior i to the protein of peanut meal. With the exception of soybean protein and soybean feed, the lype of commercial processingappeared to have little or no effect Ton the amino acid composition of the products. Acknowledgment This investigation was supported in part by funds from the iTexas Cotton Research Committee. The authors wish to express their appreciation to W. E. Sewell rof the Procter and Gamble Company and to C. W. McMath of the Traders Oil Company for their courtesy in supplying many of the samples used in this investigation. V-g-aa q-Q 12 IO. II. I2. 13. 14. 15. BULLETIN NO. 692, TEXAS AGRICULTURAL EXPERIMENT STATION Bibliography Harris, W. D., F. F. Bishop, C. M. Lyman and R. Helpert, 1947. Re- port on Isopropanol as a Solvent for Extraction of Cottonseed. J. Am. Oil Chem. Soc., V01. 24, p. 370. Kuiken, K. A., W. H. Norman, C. M. Lyman, F. Hale and L. Blotter, 1943. The Microbiological Determination of Amino Acids, I Valine, Leucine, and Isoleucine. J. Biol. Chem. Vol. 151, p. 615. Lyman, C. M., K. A. Kuiken and F. Hale, I947. The Histidine Content of Meat. J. Biol. Chem. Vol. 171, p. 233. Lyman, C. M., O. Moseley, B. Butler, S. Wood and F. Hale, 1946. The Microbiological Determination of Amino Acids, III Methionine. J. Biol. Chem. Vol. 166, p. 161. Kuiken, K. A., C. M. Lyman and F. Hale, 1947. Factors which Influ- ence the Stability of Tryptophane During Hydrolysis of Protein in Alkaline Solution. J. Biol. Chem. Vol. 1-71, p. 551. Block, R; J. and D. Bolling, 1945. The Amino Acid Composition of Proteins and Foods. Pp. 76, 136, 182, 214, 237, and 238, Spring- field. Greene, R. D. and A. Black, 1944. The Microbiological Assay of Tryp- tophane in Proteins and Foods. J. Biol. Chem. Vol. 155, p. 1. Stokes, J. L., M._Gunness, I. M. Dwyer and M. C. Caswell, 1945. Micro- biological Methods for the Determination of Amino Acids, II A Uni- form Assay for the Ten Essential Amino Acids. J. Biol. Chem. Vol. 16o, p. 35. Wooley, J. G. and W. H. Sebrell, 1945. Two Microbiological Methods for the Determination of L(-) Tryptophane in Proteins and other Complex Substances. J. Biol. Chem. Vol. 157, p. 141. Baumgarten, W., A. N. Mather and L. Stone, 1946. Essential Amino Acid Composition of Feed Materials. Cereal Chem. Vol. 23, p. 135. Greenhut, I. T., B. S. Schweigert and C. A. Elvehjem, 1946. Hydroly- sis Procedures for the Determination of Tryptophane in Proteins and Foodstuffs by the Microbiological Procedure. J. Biol. Chem. Vol. 165, p- s25. .- Greenhut, I. T., B. S. Schweigert and C. A. Elvehjem, 1946. The Amino Acid Requirements of Streptococcus F aecalis and the Use of this Organism for the Determination of Threonine in Natural Prod- ucts. J. Biol. Chem. Vol. 162, p. 69. Riesen, W. H., B. S. Schweigert and C. A. Elvehjem, 1946. Micro- biological Determination of Methionine in Proteins and F oodstufis. J. Biol. Chem. Vol. 165, p. 347. Schweigert, B. S., 1947. Amino Acid Content of Feeds, I. Leucine, Valine, Isoleucine, and Phenylalanine. J. Nutrition. Vol. 33, p. 553. Horn, M. J., D. B. Jones and A. E. Blum, 1947. Microbiological De- termination of Threonine in Proteins and Foods. J. Biol. Chem. Vol. 169, p- 739-