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Influence of Dietary Protein on Amino Acid Composition of Egg Shells1
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RESEARCH NOTES Rhodes, M. B., N. Bennett and R. E. Feeney, 1959. The flavoprotein-apoprotein system of egg white. J. Biol. Chem. 234: 2054-2060. Slosberg, H. M., H. L. Hanson, G. F. Stewart and B. Lowe, 1948. Factors influencing the effects of heat treatment on the leavening powers of egg white. Poultry Sci. 27: 294-301. Tillmans, J., and K. Philippi, 1929. Tiber den Gehalt der wichtigsten Proteine der Nahrungsmittel an Kohlehydrat und iiber ein kilorimetrischen ver Fahren zur quantitativen Bestimmung von stickstoffreien Zucker in Eiweis. Biochem. Z. 215:36-60.
INFLUENCE OF DIETARY PROTEIN ON AMINO ACID COMPOSITION OF EGG SHELLS 1 J. N. BUTTS 2 AND F. E. CUNNINGHAM Dairy and Poultry Science Department, Kansas State University, Manhattan, Kansas 66506 (Received for publication February 1, 1973) ABSTRACT Egg shells were collected from eggs produced by hens fed isocaloric rations containing 12, 14, 16, or 18% protein. The membranes were removed and the shells were acid hydrolyzed and analyzed in a Beckman Model 120 C Amino Acid Analyzer. Duplicate runs were averaged. Our results showed that protein level of the ration had no influence on quantitative or qualitative makeup of the amino acids in egg shells. POULTRY SCIENCE 52:1662-1663, 1973
INTRODUCTION
Baker and Balch (1962) found that egg shell matrix protein was characterized by the absence of hydroxyproline, by low concentration of aromatic and sulfur-containing amino acids, and by the presence of about twice as many dicarboxylic amino acids as basic amino acids. Tyler (1961) found no relationship between egg shell nitrogen and shell thickness, however. Tyler and Geake (1958) observed that true shell nitrogen and its components (soluble and insoluble nitro1 Contribution No. 857, Department of Dairy and Poultry Science, Kansas Agricultural Experiment Station, Manhattan, Kansas 66506. 2 Present address: Poultry Science Department, Purdue University, Lafayette, Indiana 47907.
gen) decreased as egg shell thickness increased. Maesso et al. (1969), who analyzed the acid hydrolyzate of inner and outer shell membranes and the cuticle, found that the cuticle was high in glycine, aspartic, and glutamic acids but low in methionine, proline, and tyrosine, and that the inner and outer membranes were similar except that arginine and cystic acid were higher in the outer membrane while proline was higher in the inner membrane. Frank et al. (1965), who analyzed the amino acid content of alkaliextracted, acid-hydrolyzed egg shell membranes, reported no major quantitative differences in amino acids between shells exhibiting high and those exhibiting low breaking strength.
Downloaded from http://ps.oxfordjournals.org/ at Purdue University Libraries ADMN on June 19, 2015
REFERENCES Cunningham, F. E., 1970. The effect of heat on egg white. World's Poultry Sci. J. 26: 783-786. Fredericq, E., and H. F. Deutsch, 1949. Studies on ovomucoid. J. Biol. Chem. 181: 499-507. Lineweaver, H., and C. W. Murray, 1947. Identification of the trypsin inhibitor of egg white with ovomucoid. J. Biol. Chem. 171: 565-582. Nath, K. R., D. V. Vadehra and R. C. Baker, 1972. Characterization of the exudate from cooked shell eggs. J. Agr. Food Chem. 20: 792-794. Nelson, N., 1944. A photometric adaptation of the Somogyi method for the determination of glucose. J. Biol. Chem. 153: 375-380.
1663
RESEARCH NOTES TABLE 1.—The effect of dietary protein level* on the amino acid composition** of egg shells 12%
14%
Lys ffis NH 3 Arg Asp Thr Ser Glu Pro Gly Ala Val Met He Leu Tyr Phe
222.5 452.4 742.2 431.5 600.0 442.8 470.8 772.3 703.0 680.0 244.4 548.8 223.5 221.1 302.3 66.8 74.4
204.1 343.2 631.8 490.7 510.1 ' 359.3 395.0 664.4 582.2 587.3 234.2 468.8 190.7 184.1 276.1 69.6 74.1
16%
18%
218.3 232.2 433.8 433.3 598.9 656.9 521.9 510.6 604.8 647.3 432.1 . 414.1 465.5 465.2 784.3 773.4 682.9 648.7 689.7 714.7 263.7 300.5 551.0 545.0 223.6 220.0 223.0 233.7 322.5 352.4 77.1 91.1 79.4 95.3
* The dietary treatment consisted of four isocalorical, pelleted laying rations with the approximate protein content of 12, 14, 16, or 18%. ** Micromoles of amino acid/gram organic matrix.
We examined the amino acid content of egg shells produced by hens maintained on diets differing in protein. METHODS AND MATERIALS Eggs used in this study came from hens being fed an isocaloric diet with a protein level of 12, 14, 16, or 18% (Butts and Cunningham, 1972). After removing the inner and outer egg shell membranes, we placed the shell in 10% HC1 at room temperature (26°C). During digestion the shell was agitated; the remaining pellicle washed in distilled HOH, then air dried overnight. Duplicate samples (14 to 15 mg.) were hydrolyzed with 6N HC1 in evacuated tubes for 24 hours at 110°C. Prior to analysis, the HC1 was evaporated under vacuum and the hydrolyzate rehydrated with citrate buffer. To determine amino
RESULTS AND DISCUSSION Data presented in Table 1 show the amino acid content of egg shells from the four dietary treatments. No significant relationship was found between increased protein in diet and amino acid content of the shells. We reported elsewhere that the 12% ration was deficient in methionine and cystine and that egg production and egg size were less for birds receiving that ration (Butts and Cunningham, 1972). This study indicates that the amino acid composition of egg shells is not affected by the amount of dietary protein, even when certain essential amino acids are limited in the ration. REFERENCES Baker, J. R., and D. A. Balch, 1962. A study of the organic material of hens' egg shell. Biochem. J. 82:352-361. Butts, J. N., and F. E. Cunningham, 1972. Effect of dietary protein on selected properties of the egg. Poultry Sci. 51:1726-1734. Frank, F. R., R. E. Burger and M. H. Swanson, 1965. The relationships among shell membrane, selected chemical properties, and the resistance to shell failure of Gattus domesticus eggs. Poultry Sci. 44: 63-69. Maesso, E. R., D. V. Vadehra and R. C. Baker, 1969. Chemical composition of egg exterior structures. Poultry Sci. 48: 1838. Tyler, C , 1961. Studies on egg shells. XVII. Variations in membrane thickness and in true shell nitrogen over different parts of the same shell. J. Sci. Food Agric. 12: 470-475. Tyler, C , and F. H. Geake, 1958. Studies on egg shells, IX. The influence of individuality, breed, and season on certain characteristics of egg shells from pullets. J. Sci. Food Agric. 9: 473-483.
Downloaded from http://ps.oxfordjournals.org/ at Purdue University Libraries ADMN on June 19, 2015
Dietary protein
acids present, a Beckman Model 120C Amino Acid Analyzer was used according to the procedure outlined in the operations manual.
RESEARCH NOTES Rhodes, M. B., N. Bennett and R. E. Feeney, 1959. The flavoprotein-apoprotein system of egg white. J. Biol. Chem. 234: 2054-2060. Slosberg, H. M., H. L. Hanson, G. F. Stewart and B. Lowe, 1948. Factors influencing the effects of heat treatment on the leavening powers of egg white. Poultry Sci. 27: 294-301. Tillmans, J., and K. Philippi, 1929. Tiber den Gehalt der wichtigsten Proteine der Nahrungsmittel an Kohlehydrat und iiber ein kilorimetrischen ver Fahren zur quantitativen Bestimmung von stickstoffreien Zucker in Eiweis. Biochem. Z. 215:36-60.
INFLUENCE OF DIETARY PROTEIN ON AMINO ACID COMPOSITION OF EGG SHELLS 1 J. N. BUTTS 2 AND F. E. CUNNINGHAM Dairy and Poultry Science Department, Kansas State University, Manhattan, Kansas 66506 (Received for publication February 1, 1973) ABSTRACT Egg shells were collected from eggs produced by hens fed isocaloric rations containing 12, 14, 16, or 18% protein. The membranes were removed and the shells were acid hydrolyzed and analyzed in a Beckman Model 120 C Amino Acid Analyzer. Duplicate runs were averaged. Our results showed that protein level of the ration had no influence on quantitative or qualitative makeup of the amino acids in egg shells. POULTRY SCIENCE 52:1662-1663, 1973
INTRODUCTION
Baker and Balch (1962) found that egg shell matrix protein was characterized by the absence of hydroxyproline, by low concentration of aromatic and sulfur-containing amino acids, and by the presence of about twice as many dicarboxylic amino acids as basic amino acids. Tyler (1961) found no relationship between egg shell nitrogen and shell thickness, however. Tyler and Geake (1958) observed that true shell nitrogen and its components (soluble and insoluble nitro1 Contribution No. 857, Department of Dairy and Poultry Science, Kansas Agricultural Experiment Station, Manhattan, Kansas 66506. 2 Present address: Poultry Science Department, Purdue University, Lafayette, Indiana 47907.
gen) decreased as egg shell thickness increased. Maesso et al. (1969), who analyzed the acid hydrolyzate of inner and outer shell membranes and the cuticle, found that the cuticle was high in glycine, aspartic, and glutamic acids but low in methionine, proline, and tyrosine, and that the inner and outer membranes were similar except that arginine and cystic acid were higher in the outer membrane while proline was higher in the inner membrane. Frank et al. (1965), who analyzed the amino acid content of alkaliextracted, acid-hydrolyzed egg shell membranes, reported no major quantitative differences in amino acids between shells exhibiting high and those exhibiting low breaking strength.
Downloaded from http://ps.oxfordjournals.org/ at Purdue University Libraries ADMN on June 19, 2015
REFERENCES Cunningham, F. E., 1970. The effect of heat on egg white. World's Poultry Sci. J. 26: 783-786. Fredericq, E., and H. F. Deutsch, 1949. Studies on ovomucoid. J. Biol. Chem. 181: 499-507. Lineweaver, H., and C. W. Murray, 1947. Identification of the trypsin inhibitor of egg white with ovomucoid. J. Biol. Chem. 171: 565-582. Nath, K. R., D. V. Vadehra and R. C. Baker, 1972. Characterization of the exudate from cooked shell eggs. J. Agr. Food Chem. 20: 792-794. Nelson, N., 1944. A photometric adaptation of the Somogyi method for the determination of glucose. J. Biol. Chem. 153: 375-380.
1663
RESEARCH NOTES TABLE 1.—The effect of dietary protein level* on the amino acid composition** of egg shells 12%
14%
Lys ffis NH 3 Arg Asp Thr Ser Glu Pro Gly Ala Val Met He Leu Tyr Phe
222.5 452.4 742.2 431.5 600.0 442.8 470.8 772.3 703.0 680.0 244.4 548.8 223.5 221.1 302.3 66.8 74.4
204.1 343.2 631.8 490.7 510.1 ' 359.3 395.0 664.4 582.2 587.3 234.2 468.8 190.7 184.1 276.1 69.6 74.1
16%
18%
218.3 232.2 433.8 433.3 598.9 656.9 521.9 510.6 604.8 647.3 432.1 . 414.1 465.5 465.2 784.3 773.4 682.9 648.7 689.7 714.7 263.7 300.5 551.0 545.0 223.6 220.0 223.0 233.7 322.5 352.4 77.1 91.1 79.4 95.3
* The dietary treatment consisted of four isocalorical, pelleted laying rations with the approximate protein content of 12, 14, 16, or 18%. ** Micromoles of amino acid/gram organic matrix.
We examined the amino acid content of egg shells produced by hens maintained on diets differing in protein. METHODS AND MATERIALS Eggs used in this study came from hens being fed an isocaloric diet with a protein level of 12, 14, 16, or 18% (Butts and Cunningham, 1972). After removing the inner and outer egg shell membranes, we placed the shell in 10% HC1 at room temperature (26°C). During digestion the shell was agitated; the remaining pellicle washed in distilled HOH, then air dried overnight. Duplicate samples (14 to 15 mg.) were hydrolyzed with 6N HC1 in evacuated tubes for 24 hours at 110°C. Prior to analysis, the HC1 was evaporated under vacuum and the hydrolyzate rehydrated with citrate buffer. To determine amino
RESULTS AND DISCUSSION Data presented in Table 1 show the amino acid content of egg shells from the four dietary treatments. No significant relationship was found between increased protein in diet and amino acid content of the shells. We reported elsewhere that the 12% ration was deficient in methionine and cystine and that egg production and egg size were less for birds receiving that ration (Butts and Cunningham, 1972). This study indicates that the amino acid composition of egg shells is not affected by the amount of dietary protein, even when certain essential amino acids are limited in the ration. REFERENCES Baker, J. R., and D. A. Balch, 1962. A study of the organic material of hens' egg shell. Biochem. J. 82:352-361. Butts, J. N., and F. E. Cunningham, 1972. Effect of dietary protein on selected properties of the egg. Poultry Sci. 51:1726-1734. Frank, F. R., R. E. Burger and M. H. Swanson, 1965. The relationships among shell membrane, selected chemical properties, and the resistance to shell failure of Gattus domesticus eggs. Poultry Sci. 44: 63-69. Maesso, E. R., D. V. Vadehra and R. C. Baker, 1969. Chemical composition of egg exterior structures. Poultry Sci. 48: 1838. Tyler, C , 1961. Studies on egg shells. XVII. Variations in membrane thickness and in true shell nitrogen over different parts of the same shell. J. Sci. Food Agric. 12: 470-475. Tyler, C , and F. H. Geake, 1958. Studies on egg shells, IX. The influence of individuality, breed, and season on certain characteristics of egg shells from pullets. J. Sci. Food Agric. 9: 473-483.
Downloaded from http://ps.oxfordjournals.org/ at Purdue University Libraries ADMN on June 19, 2015
Dietary protein
acids present, a Beckman Model 120C Amino Acid Analyzer was used according to the procedure outlined in the operations manual.