Proteins are complex, organic compounds composed of many amino acids linked together through peptide bonds and

cross-linked between chains by sulfhydryl bonds, hydrogen bonds and van der Waals forces Proteins, from the Greek proteios, meaning first, are a class of organic compounds which are present in and vital to every living cell

In the form of skin, hair, callus, cartilage, muscles, tendons and ligaments, proteins hold together, protect, and provide structure to the body of a multi-celled organism. In the form of enzymes, hormones, antibodies, and globulins, they catalyze, regulate, and protect the body chemistry. In the form of hemoglobin, myoglobin and various lipoproteins, they effect the transport of oxygen and other substances within an organism

Properties
Proteins can also be characterized by their chemical reactions. Most proteins are soluble in water, in alcohol, in dilute base or in various concentrations of salt solutions Proteins have the characteristic coiled structure which is determined by the sequence of amino acids in the primary polypeptide chain and the stereo configuration of the radical groups attached to the alpha carbon of each amino acid Proteins are heat labile exhibiting various degrees of lability depending upon type of protein, solution and temperature profile

Proteins can be reversible or irreversible, denatured by heating, by salt concentration, by freezing, by ultrasonic stress or by aging. Proteins undergo characteristic bonding with other proteins in the socalled plastein reaction and will combine with free aldyhyde and hydroxy groups of carbohydrates to form Maillard type compounds.

Classification
Proteins can be classified as: (a) Simple proteins. On hydrolysis they yield only the amino acids and occasional small carbohydrate compounds. Examples are: albumins, globulins, glutelins, albuminoids, histones and protamines.

b) Conjugated proteins. These are simple proteins combined with some non-protein material in the body. Examples are: nucleoproteins, glycoproteins, phosphoproteins, haemoglobins and lecithoproteins. (c) Derived proteins. These are proteins derived from simple or conjugated proteins by physical or chemical means. Examples are: denatured proteins and peptides.

1. 2. 3. 4. 5. 6. 7. 8. 9.

Histidine Isoleucine Leucine Lysine Methionine Phenylalanine Threonine Tryptophan Valine

1. Alanine 2. Arginine* 3. Aspartic acid 4. Cysteine* 5. Cystine 6. Glutamic acid 7. Glutamine* 8. Glycine 9. Proline 10. Serine 11. Tyrosine

Essential Amino Acids

Nonessential Amino Acids

Protein

Analysis: Determine total N by Kjeldahl

All N NH4+ Determine as NH3 Total N x 6.25 = crude protein

Nitrogenous Compounds in Feeds

True proteins Polymers of AA (18 to 20 AA) linked by peptide bonds
Essential AA

Have to be present in the diet (absorbed) Arg Lys Trp Leu Ile Val Met Thr Phy His PVT TIM HALL
Nonessential amino acids (dispensable)

Synthesized in body tissues Glu Gly Asp Pro Ala Ser Cys Tyr
Proteins Peptides Amino acids

Nonprotein nitrogen
Nitrogen not associated with protein
Free amino acids, nucleic acids, amines, ammonia, nitrates, nitrites, urea

Crude protein
Total nitrogen x 6.25 Proteins on average contain 16% nitrogen

Protein Degradation in the Rumen

Feed proteins Peptides Amino acids

Undegraded feed proteins Escaped feed proteins Bypass proteins

Enzymes from protozoa and bacteria Many species of bacteria involved Bacterial enzymes are extracellular Enzymes not in cell free rumen fluid Both exopeptidase and endopeptidase activity

Protein Pathways

Bypass proteins
Proteins that are not extensively degraded in the rumen 1. Natural Corn proteins, blood proteins, feather meal 2. Modification of feed proteins to make them less degradable Heat - Browning or Maillard reaction Expeller SBM, Dried DGS, Blood meal Chemical Formaldehyde Polyphenols Tannins Alcohol + heat Usually some loss in availability of amino acids - lysine

Factors Affecting Ruminal Protein Degradation

Chemical Nature of the proteins Solubility More soluble proteins degraded faster Some soluble proteins not extensively degraded Egg ovalbumin, serum proteins

3-dimensional structure Affects solubility & availability Chemical bonding Disulfide bonds Reduces degradation Physical barriers Cell walls of plants Cross linking of peptide chains Reduces degradation Aldehydes, Tannins

Feed intake Rate of passage Time proteins remain in the rumen Feed processing Rate of passage Heat damage Complexes with carbohydrates

Estimating Degradation of Dietary Proteins in the Rumen

In situ digestion Feed placed in Dacron bags suspended in the rumen Measure protein lost over time 2. Cannulated animals (rumen & duodenum) Measure protein flowing through duodenum Need to differentiate feed from microbes 3. In vitro incubation with rumen microbes Relative differences among proteins 4. In vitro digestion with fungal enzymes

Ruminant vs Nonruminant - Similarities

tissue level Metabolic pathways similar 2. Ruminant tissues can synthesize dispensable AA 3. Cannot synthesize indispensable AA
1. At

Essential AA must be provided from digestive tract

4. Tissue proteins constantly undergoing turnover 5. AA not stored 6. Constant supply of AA required

Ruminant vs Nonruminant - Dissimilaritie

1. Microbial population has profound effect on AA reaching S.I.
a. AA profile at S.I. different from diet
Up-grades low quality dietary protein Down-grades high quality dietary protein

b. Enables ruminants to use NPN

efficiently

Ruminants can be productive without a source of dietary true protein

c. Animal can survive on low amounts of

dietary protein by recycling N (as urea) back to rumen

Ruminant vs Nonruminant - Dissimilarities

2. In ruminant nutrition generally not concerned with AA composition of dietary protein

a. Type of feed does not affect AA

comp. of bacteria and protozoa leaving rumen

AA comp. of MOs reaching duodenum strikingly similar when measured in labs around the world

b. Biological value (BV) of

microbial protein ~80%

Degradation of NPN Compounds

Activity associated with microorganisms Urea CO2 + 2 NH3 High concentrations of urease activity in the rumen Low concentrations of urea in the rumen Biuret 2 CO2 + 3 NH3 Low activity in the rumen NO3 NH3

Fate of Free Amino Acids in the Rumen Amino acids not absorbed from the rumen Concentrations of free AA in the rumen very low

Amino acids and small peptides (up to 5 AA) transported into bacterial cells Na pumped out of cells Uses ATP Na gradient facilitates transport of AA by a carrier Utilized for synthesis of microbial proteins Amino acids metabolized to provide

Amino Acid Degradation in the Rumen NH3 Amino acids Keto acids CO2 VFA

Enzymes from microorganisms Intracellular enzymes Peptides probably hydrolyzed to amino acids and then degraded NH3, VFA and CO2 absorbed from rumen

High numbers Low activity

Butrivibrio fibrisolvens Measphaera elsdenii Selenomonas ruminantium 109 per ml 10 to 20 NMol NH3 per min per mg protein Monensin resistant Involved in CHOH fermentation

Low numbers High activity

Clostridium aminophilum Clostridium sticklandii Peptostreptococuss anaerobius 107 per ml 300 NMol NH3 per min per mg protein Monensin sensitive Ferment CHOH slowly or not at all

Rumen microbes
Bacteria (50% CP) Protozoa (20-60%, avg 40% CP) Bacteria major player, % of microbial N entering SI from protozoa < 10% N source for microbes Diet protein Non protein N Recycled N

Nutritive Value of Microbial N

Increases value of low quality feed N Decreases value of high quality feed N Animal can survive on non-protein N Can survive on low amounts of recycled N

Urea
Urea = 281% CP equivalent N = 45% of urea 45%N x 6.25 = 281% CP Dairy cows Upper limit ~1% of diet DM Palatability begins to limit intake Above 0.75% urea in diet DM, start observing palatability problems ( intake)

Urea Toxicity (NH3 Toxicity

Mechanism Rumen [NH3]

Rumen pH As pH , shift from NH4+ to NH3 NH3 absorbed faster than NH4+ Liver capacity to convert NH3 to urea is exceeded NH3 goes to blood 2 mg NH3/100 ml plasma is toxic

Signs of toxicity

Appear 20-30 min after urea ingestion Rapid and labored breathing Tremors Incoordination Inability to stand & tetany increasingly apparent

Treatment

Orally dose with 5% acetic acid (~1 gal. for 1,000 lb cow) Shift equilibrium from NH3 to NH4+ rate of absn Drench with cold water rumen temp. which rate of urea hydrolysis Dilutes NH3 concentration Takes 6-12 gal.; not practical when several sick

Prevention

Mix feeds well Dont switch rapidly from natural protein to urea Always have feed available Dont allow hungry animals access to highly palatable, high urea diet, feed, or supplement (including lick tanks) Dont use urea with low-energy feeds