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Characterization and Bioavailability of Alfalfa Leaf Protein Concentrate. Sirous Kashefi Louisiana State University and Agricultural & Mechanical College

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Characterization and Bioavailability of Alfalfa Leaf Protein Concentrate. Sirous Kashefi Louisiana State University and Agricultural & Mechanical College Louisiana State University LSU Digital Commons LSU Historical Dissertations and Theses Graduate School 1982 Characterization and Bioavailability of Alfalfa Leaf Protein Concentrate. Sirous Kashefi Louisiana State University and Agricultural & Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_disstheses Recommended Citation Kashefi, Sirous, "Characterization and Bioavailability of Alfalfa Leaf Protein Concentrate." (1982). LSU Historical Dissertations and Theses. 3810. https://digitalcommons.lsu.edu/gradschool_disstheses/3810 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Historical Dissertations and Theses by an authorized administrator of LSU Digital Commons. For more information, please contact [email protected]. INFORMATION TO USERS This reproduction was made from a copy of a document sent to us for microfilming. While the most advanced technology has been used to photograph and reproduce this document, the quality of the reproduction is heavily dependent upon the quality of the material submitted. The following explanation of techniques is provided to help clarify markings or notations which may appear on this reproduction. 1.The sign or “target” for pages apparently lacking from the document photographed is “Missing Page(s)” . If it was possible to obtain the missing page(s) or section, they are spliced into the film along with adjacent pages. This may have necessitated cutting through an image and duplicating adjacent pages to assure complete continuity. 2. When an image on the film is obliterated with a round black mark, it is an indication of either blurred copy because of movement during exposure, duplicate copy, or copyrighted materials that should not have been filmed. For blurred pages, a good image of the page can be found in the adjacent frame. If copyrighted materials were deleted, a target note will appear listing the pages in the adjacent frame. 3. When a map, drawing or chart, etc., is part of the material being photographed, a definite method of “sectioning” the material has been followed. It is customary to begin filming at the upper left hand comer of a large sheet and to continue from left to right in equal sections with small overlaps. If necessary, sectioning is continued again—beginning below the first row and continuing on until complete. 4. For illustrations that cannot be satisfactorily reproduced by xerographic means, photographic prints can be purchased at additional cost and inserted into your xerographic copy. These prints are available upon request from the Dissertations Customer Services Department. 5. Some pages in any document may have indistinct print. In all cases the best available copy has been filmed. University Microfilms International 300 N. Zeeb Road Ann Arbor, Ml 48106 8312093 Kashefi, Sirous CHARACTERIZATION AND BIOAVAILABILITY OF ALFALFA LEAF PROTEIN CONCENTRATE The Louisiana State University and Agricultural and Mechanical Ph.D. Col1982 University Microfilms International300 N. Zeeb Road, Ann Arbor, MI 48106 CHARACTERIZATION AND BIOAVAILABILITY OF ALFALFA LEAF PROTEIN CONCENTRATE A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural a:.d I'echanical College in partial fulfillment of the requirements for the Degree of Doctor of Philosophy in The Department of Food Science by Sirous Kashefi B.S. College of Food Science and Nutrition, Iran, 1972 M.A. Texas Southern University, 1978 December, 1982 ACKNOWLEDGEMENT The author is Indebted and deeply grateful to his major professor, Dr. Fred H. Hoskins, for his encouragement and guidance throughout the course of this study. Appreciation is also extended to Dr. J.A. Liuzzo, for his advice and encouragement during the early phases of this research. Acknowledgements are also extended to Dr. A.M. Mullins, Dr. R.M Grodner, and Dr. R.H. Gough for serving on his Advisory Committee. The author would like to express sincere appreciation to his father, mother, sister and brothers for their encouragement and sacrifice, without which this work could not have been completed. Finally, the author would also wish to express his appreciation to Linda, for her great help and encouragement. ii TABLE OF CONTENTS Page ACKNOWLEDGEMENTS.............................................. ii LIST OF TABLES............................................... v LIST OF FIGURES............................................... vi ABSTRACT..................................................... vii INTRODUCTION.................................................. 1 LITERATURE REVIEW.............................. 4 The Extraction of Leaf Protein Concentrates.............. 4 Preservation of Leaf Protein Concentrates................ 6 Source of Leaf Protein................................... 8 The Nutritive Composition of Leaf ProteinConcentrate 10 Methods of Evaluation of the Protein Quality of LPC’s 13 Tetrahymena pyriformis Assay........................... 13 Animal Feeding Studies................................. 16 Human Studies.......................................... 18 The Chemical Nature of Leaf Protein Concentrates...... 19 Leaf Protein Concentrate as an Ingredient in Food Formulations........... 22 MATERIALS AND METHODS.......................................... 25 Sampling...................... 25 Protein Extraction....................... .............. 25 Nitrogen Determination................................... 27 Moisture Determination................................... 27 Fat Determination................. 27 Amino Acid Analysis...................................... 27 Protein Solubilization................... .............. 27 iii TABLE OF CONTENTS (cont'd.) Page Relative Nutritive Value (RNV) Determination.............. 28 Measuring of Growth.of TpW............................ 29 Electrophoresis of the Protein (Molecular Weight Determination) ...................................... 32 Preparation of Protein Solution....................... 32 Preparation of Samples................... 33 Electrophoretic Separation............................. 33 Evaluation of Meat Loaves Containing Soy and Alfalfa Leaf Protein........................................... 34 Sensory Evaluation....................................... 35 RESULTS AND DISCUSSION........................................ 36 ALPC Analysis............................................ 36 Relative Nutritive Value................................. 40 Solubilization of ALPC................................... 44 Electrophoretic Behavior of ALPC......................... 47 Sensory Evaluation of Extended Ground Beef......... 58 Flavor................................................. 58 Odor........................................... 58 Color.................................................. 58 Tenderness............................................. 58 Juiciness.............................................. 59 Appearance............................................. 59 Overall Acceptability.................................. 59 SUMMARY AND CONCLUSIONS............ 61 LITERATURE CITED.............................................. 64 APPENDICES.................................................... 76 VITA.......................................................... 85 iv LIST OF TABLES Table Page 1. Amino acid composition of alfalfa LPC........................ 12 2. Proximate analysis of chloroplastic and cytoplasmic fractions of ALPC.......................... 37 3. Amino acid composition of chloroplastic and cytoplasmic fractions of ALPC.................. 38 4. Chemical scores for cytoplasmic fraction of APLC, compared to several standard proteins............................. 39 5. Quantitative requirements of Tetrahymena pyriformls for essential amino acids........ 41 6. Cell density of Tetrahymena pyriformls of casein, soy protein, cytoplasmic and chloroplastic fractions of ALPC ......... 42 7. Relative nutritive value (RNV) and Tetrahymena PER of soy protein, cytoplasmic and chloroplastic fractions of ALPC.. 43 8. ALPC solubilized at different pH’s in the presence of 0.01M sodium phosphate, 1% 6-mercaptoethanol and 1% SDS........ 45 9. Relative percent and concentration of alfalfa juice protein subunits.using SDS-PAGE and photodensitometry....... 50 10. Relative percent and concentration of protein subunits in alfalfa before and after freeze drying................... 51 11. Sensory evaluation of ground beef extended with soy and alfalfa leaf protein..................................... 60 v LIST OF FIGURES Figure Page 1. Preparation of fractionated alfalfa leaf protein concentrate (APLC).................... 26 2. Tetrahymena pyriformis W. bioassay procedure............... 30 3. Procedure for RNV determination.................. 31 4. Percent ALPC solubilized as a function of pH in a 0.01M sodium phosphate buffer solution containing SDS (1%) and 6-mercaptoethanol (1%)................................. 46 5. Electrophoretograms of protein in alfalfa before and after extraction............................................. 49 6. Photodensitometric tracing of alfalfa juice protein........ 52 7. Comparison of photodensitometric tracing of alfalfa juice protein and alfalfa protein concentrate before freeze drying................................ 53 8. Comparison of photodensitometric tracing of alfalfa juice protein and alfalfa protein concentrate after freeze drying................................................. 54 9. Comparison of photodensitometric
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