Application of Bromelain Extract for Muscle
Foods Tenderization
by Sunantha Ketnawa, Saroat Rawdkuen
INTRODUCTION
PROBLEM STATEMENT
Bromelain is an enzyme extract from the fresh pineapples. "Bromelain" term refer to enzymes extract from the plants of the family Bromeliaceae, or it refer to a combination of the enzymes along with other compounds produced in an extract. Fruit bromelain and stem bromelain also other name of bromelain enzymes.
Bromelain present in all parts of the pineapple.Bromelain is a white to tan powder, soluble in water, and is stable at temperatures of 50–60 °C.Cooked or canned pineapple will not have a tenderizing effect, because the bromelain enzymes already denatured in high temperature.
METHODS
- Determination of pH
- Determination of Moisture Content
- Determination of Water-holding Capacity
- TCA-Soluble Peptides Content
-Meat become TOUGHNESS causing of amount intramuscular connective tissue, intramuscular fat and the length of the sarcomere
-Cow that already slaughter make their meat become toughness because the cow stress during the process slaughtering
-Using mechanical tool to make the meat tender become more difficult to handle.
RESULT
DISCUSSION
CONCLUSION
In this research study, it stated that bromelain act as enzyme that functioning in tenderizing meat protein structure, these experiment are the evidences of bromelain act as tenderizing agent. There are two type of properties that tested which is physically and chemically properties on beef, chicken and squid meat. For physically properties, it determined several test, first one is textural properties where shear force were taken place in this test, the result show the shear force value decrease when level concentration BE increased.
Effect of Bromelain Extract on the Physical Properties of Muscle Foods
In summary, bromelain extract can be good in tenderizing meat and easy to extract because can take from fresh pineapple but not from canned or cooked pineapple.This application of bromelain extract for muscle foods tenderization play significance role to meat based foods industry to use bromelain for tenderizing meat.It will save more time and can increase production of product.We hope that in future there will be more efficient way to tenderize meat using more advance technology,so that can save time and money but can produce more product.
- Texture Properties
- Water Holding Capacity and Cooking Yield
- Muscle Microstructure
Effect of Bromelain Extract on the Chemical Properties of Muscle Foods
- pH and Moisture Content
- TCA-Soluble Peptides Content
- Hydrolytic Patterns
GROUP MEMBERS
METHODS
Determination of pH Approximately 2 g of meat was taken in 4 test tubes. The tubes were incubated with 1%, 2%, 3% and 4% solutions of Bromelain in distilled water and a test tube without Bromelain, served as control. It was allowed to stand for 10 minutes and was homogenized by grinding followed by crushing mechanically using a mortar and pestle. The pH values were determined using a digital pH meter. The pH meter was calibrated using a standard sodium acetatebuffer of pH 4 and the pH of the samples was measured.
Determination of Moisture Content
5 g of meat (treated with, 1%, 2%, 3% and 4% of Bromelain) was suspended in 10 mL of 0.5 M sodium chloride (NaCl) solution for 20 minutes. The weight of the meat was noted. They were placed on a glass plate and heated at 100°C for 20 minutes using a hot plate. The weight of the meat after removing moisture was determined. The tube then was held at 4° C for 15 min, stirred again for 1 min and centrifuged at 10,000 rpm at 4° C for 15 min. After centrifugation, the volume of the supernatant was measured and the water-holding capacity of the meat treated with various concentrations of meat.
Determination of Water-holding Capacity
Water-holding capacity (WHC) was determined according to the procedure described by Wardlaw.
11 6 g samples of meat with 1%, 2%, 3% and 4% of Bromelain were stirred for 1 min with 10 ml of 0.6 M sodium chloride (NaCl) solution in a 15 ml centrifuge tube.
TCA-Soluble Peptides Content
The highest content of TCA-soluble peptides was found in the chicken, followed by beef muscle treated with 30%. When the concentration of bromelain extract was increased, the TCA-soluble peptides content in the entire treated sample increased (0.05). The TCA-soluble peptides content was the low test in the beef muscle. This result coordinates with the protein patterns that show higher intensity for major protein bands. More than a 25% increase of TCA-soluble peptides content was found in the sample treated with 20% bromelain extract as compared to the control. The highest increase of TCA-soluble peptides content was found in the chicken and the lowest was found in the beef muscle. The tyrosine level indicated that the endogenous oligopeptides and free amino acids, as well as degradation products, all accumulated after being marinated with bromelain extract .
PHYSICAL PROPERTIES
Textural Properties
The toughness is indicated by the amount of intramuscular connective tissue, intramuscular fat and the length of the sarcomere. Tenderization associated with postmortem aging is due to endogenous enzyme activity to the muscle. The toughness of the meat samples are presented in Page 4
Water Holding Capacity and Cooking Yield
There are a significant reduction of Water Holding Capascity in all of the treated sample. The reduction in Water Holding Capacity in all of the treated samples might be due to lower pH. This drop in pH may be responsible for the overall reduction in protein reactive groups available for water holding. The lowered Water Holding Capacity in the treated samples also probably due to the denaturation of myofibrillar proteins, which play a role in water retention.
Muscle Microstructure
The control samples of all the muscle fibers had well organized structure and were closely bound to each other. Wherease in the BE treated samples, there were broken muscle fibers in different bundles, they were less attached, and there was a loss of muscle fibers interaction. The result showed strong muscle fibers were broken and the cell membranes were severely degraded. The junctions of the myofibrillar package and the sarcolemma have disappeared from most cells.
Graph Concentration vs Toughness
Toughness (N.Sec)
Bromelin Extract Concentration (%, w/w)
CHEMICAL PROPERTIES
pH and Moisture Content
Significant decrease in pH was found in all of the treated samples when compared to the control. The low pH of BE was probably caused by the lower pH of the treated samples. The powder of bromelain extract could absorb more water in the fresh meat samples, which resulted in lower moisture content when compared with the control (p< 0.05). The moisture content in the BE-treated samples markedly when decreased compared with the untreated samples. Increased BE concentration resulted in moisture content reduction, justified that enzyme treatment improved hydrophilic properties.
TCA-Soluble Peptides Content
The highest content of TCA-soluble peptides was found in the chicken, followed by beef and squid muscle treated with 20% BE (w/w) respectively. The movement of concentration of BE is parallel with the movement of the TCA-soluble peptides as shown as in the page 6.
Hydrolytic Patterns
The myosin heavy chain (MHC) and acetin (AC) are the major proteins in all muscle foods. Apart from MHC and AC, a new protein band with high intensity, paramyosin(PM), was observed in the protein petterns of the squid muscle. The degradation of muscle protein also plays a major role in determining the tenderness and Water Holding Capacity of meat during the postmortem storage. Myofibrillar proteins such as troponin-T, titin and nebulin are considered to be indicators of the overall extent of postmortem proteolysis and predictors of meat tenderness.Result showed it could be determined that BE potentially has high proteolytic activity, making the meat more tender.
GRAPH CONCENTRATION vs TCA SOLUBLE PEPTIDES
TCA soluble peptides
Bromelain Extract Concentration (%, w/w)
Discussion
In this research study, it stated that bromelain act as enzyme that functioning in tenderizing meat protein structure, these experiment are the evidences of bromelain act as tenderizing agent. There are two type of properties that tested which is physically and chemically properties on beef, chicken and squid meat. For physically properties, it determined several test, first one is textural properties where shear force were taken place in this test, the result show the shear force value decrease when level concentration BE increased. The meat firmness causes of action proteolytic on myofibrillar protein. Second test are, water holding capacity and cooking yield, the result have been observed that there is significant reduction of WHC in all treated sample. The level of WHC may effect of pH level of meat, because pH value may responsible for overall reduction. There are three main factors involved in the shrinkage or swelling of the myofibrils, the onset of rigor mortis, the decline in pH and protein fragmentation. For cooking yield, the treated sample shown significantly different compared to control sample and marinated BE sample show reduction in cooking yield. This result indicate that the thermal treatment could remove water form treated. Reduction of cooking yield may cause by degradation of sarcoplasmic and myofibrillar proteins. Lastly are muscle microstructure test, based on the result, control sample shown the muscle fiber had well organized structure and closely bound each other compared to treated sample. This may due degradation of endomysial collagen and sarcolemma surrounding the muscle fibers also it show evident the degredation of muscle fiber occur where the fiber brokem and cell membrane degraded.
For chemically properties experiment, first experiment have been carried out are pH and moisture content, there is significant different which is decrease in pH compare to control sample. The pH value in meat product is highly important because it has a major influence on other physic-chemical and quality properties such as WHC, tenderness and juiciness. Changes in pH also may due to the postmortem metabolism and application added substance to meat. For the Aw, bromelain powder extract on treated sample can absorb more water in fresh meat, automatically causing low Aw compared to control sample. The increased of BE concentration, the more reduction of Aw on meat, It is also probably due to the hygroscopic nature of BE powder when attached to the wet surface of the sample. The low moisture content of the treated samples is related to product acceptability for the consumer. Second experiment, TCA-soluble peptide, the hypothesis is the increase the concentration of BE, the highest TCA – soluble peptide in protein structure. From the result, the highest TCA – soluble content indicates greater muscle protein hydrolysis generated by proteolytic enzyme in BE. Last experiment is hydrolytic pattern, the myosin heavy chain (MHC) and actin (AC) are the major proteins in all muscle foods. A new protein band with high intensity, paramyosin (PM), beside (MHC) and (AC), observed in the protein patterns of the squid muscle. When comparing the treatment to the control, degradation of the AC and the MHC band was markedly degraded into lower molecular weight products. These enzymes have very broad specificities, and therefore unexpected break down the major muscle proteins where can give result on over – tenderization. During the postmortem storage, the degradation of muscle protein occur in meat which plays a major role in determining the tenderness and WHC. Indicators of the overall extent of postmortem proteolysis and predictors of meat tenderness are based on myofibrillar proteins. So it could be determined that BE potentially has high proteolytic activity, making the meat more tender.
GROUP MEMBERS
1. AHMAD SYAKIR BIN AHAMAD AZAHARI (F19B0010)2. RAJA ZAFRAN AIMAN BIN RAJA SALLEH (F19A0176) 3. MOHAMAD AIMAN SYAKIR BIN MOHAMAD SAITY (F19B0064) 4. MUHAMMAD MUAZ BIN MEFTAHUDDIN (F19B0265) 5. MUHAMMAD SYAHIRZAMIE BIN SHAHRUM (F19B0268)
Albert Einstein
Bichemistry Poster
Ahmad Syakir
Created on June 12, 2020
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Transcript
Application of Bromelain Extract for Muscle Foods Tenderization
by Sunantha Ketnawa, Saroat Rawdkuen
INTRODUCTION
PROBLEM STATEMENT
Bromelain is an enzyme extract from the fresh pineapples. "Bromelain" term refer to enzymes extract from the plants of the family Bromeliaceae, or it refer to a combination of the enzymes along with other compounds produced in an extract. Fruit bromelain and stem bromelain also other name of bromelain enzymes. Bromelain present in all parts of the pineapple.Bromelain is a white to tan powder, soluble in water, and is stable at temperatures of 50–60 °C.Cooked or canned pineapple will not have a tenderizing effect, because the bromelain enzymes already denatured in high temperature.
METHODS
-Meat become TOUGHNESS causing of amount intramuscular connective tissue, intramuscular fat and the length of the sarcomere -Cow that already slaughter make their meat become toughness because the cow stress during the process slaughtering -Using mechanical tool to make the meat tender become more difficult to handle.
RESULT
DISCUSSION
CONCLUSION
In this research study, it stated that bromelain act as enzyme that functioning in tenderizing meat protein structure, these experiment are the evidences of bromelain act as tenderizing agent. There are two type of properties that tested which is physically and chemically properties on beef, chicken and squid meat. For physically properties, it determined several test, first one is textural properties where shear force were taken place in this test, the result show the shear force value decrease when level concentration BE increased.
Effect of Bromelain Extract on the Physical Properties of Muscle Foods
In summary, bromelain extract can be good in tenderizing meat and easy to extract because can take from fresh pineapple but not from canned or cooked pineapple.This application of bromelain extract for muscle foods tenderization play significance role to meat based foods industry to use bromelain for tenderizing meat.It will save more time and can increase production of product.We hope that in future there will be more efficient way to tenderize meat using more advance technology,so that can save time and money but can produce more product.
Effect of Bromelain Extract on the Chemical Properties of Muscle Foods
GROUP MEMBERS
METHODS
Determination of pH Approximately 2 g of meat was taken in 4 test tubes. The tubes were incubated with 1%, 2%, 3% and 4% solutions of Bromelain in distilled water and a test tube without Bromelain, served as control. It was allowed to stand for 10 minutes and was homogenized by grinding followed by crushing mechanically using a mortar and pestle. The pH values were determined using a digital pH meter. The pH meter was calibrated using a standard sodium acetatebuffer of pH 4 and the pH of the samples was measured. Determination of Moisture Content 5 g of meat (treated with, 1%, 2%, 3% and 4% of Bromelain) was suspended in 10 mL of 0.5 M sodium chloride (NaCl) solution for 20 minutes. The weight of the meat was noted. They were placed on a glass plate and heated at 100°C for 20 minutes using a hot plate. The weight of the meat after removing moisture was determined. The tube then was held at 4° C for 15 min, stirred again for 1 min and centrifuged at 10,000 rpm at 4° C for 15 min. After centrifugation, the volume of the supernatant was measured and the water-holding capacity of the meat treated with various concentrations of meat. Determination of Water-holding Capacity Water-holding capacity (WHC) was determined according to the procedure described by Wardlaw. 11 6 g samples of meat with 1%, 2%, 3% and 4% of Bromelain were stirred for 1 min with 10 ml of 0.6 M sodium chloride (NaCl) solution in a 15 ml centrifuge tube. TCA-Soluble Peptides Content The highest content of TCA-soluble peptides was found in the chicken, followed by beef muscle treated with 30%. When the concentration of bromelain extract was increased, the TCA-soluble peptides content in the entire treated sample increased (0.05). The TCA-soluble peptides content was the low test in the beef muscle. This result coordinates with the protein patterns that show higher intensity for major protein bands. More than a 25% increase of TCA-soluble peptides content was found in the sample treated with 20% bromelain extract as compared to the control. The highest increase of TCA-soluble peptides content was found in the chicken and the lowest was found in the beef muscle. The tyrosine level indicated that the endogenous oligopeptides and free amino acids, as well as degradation products, all accumulated after being marinated with bromelain extract .
PHYSICAL PROPERTIES
Textural Properties The toughness is indicated by the amount of intramuscular connective tissue, intramuscular fat and the length of the sarcomere. Tenderization associated with postmortem aging is due to endogenous enzyme activity to the muscle. The toughness of the meat samples are presented in Page 4 Water Holding Capacity and Cooking Yield There are a significant reduction of Water Holding Capascity in all of the treated sample. The reduction in Water Holding Capacity in all of the treated samples might be due to lower pH. This drop in pH may be responsible for the overall reduction in protein reactive groups available for water holding. The lowered Water Holding Capacity in the treated samples also probably due to the denaturation of myofibrillar proteins, which play a role in water retention. Muscle Microstructure The control samples of all the muscle fibers had well organized structure and were closely bound to each other. Wherease in the BE treated samples, there were broken muscle fibers in different bundles, they were less attached, and there was a loss of muscle fibers interaction. The result showed strong muscle fibers were broken and the cell membranes were severely degraded. The junctions of the myofibrillar package and the sarcolemma have disappeared from most cells.
Graph Concentration vs Toughness
Toughness (N.Sec)
Bromelin Extract Concentration (%, w/w)
CHEMICAL PROPERTIES
pH and Moisture Content Significant decrease in pH was found in all of the treated samples when compared to the control. The low pH of BE was probably caused by the lower pH of the treated samples. The powder of bromelain extract could absorb more water in the fresh meat samples, which resulted in lower moisture content when compared with the control (p< 0.05). The moisture content in the BE-treated samples markedly when decreased compared with the untreated samples. Increased BE concentration resulted in moisture content reduction, justified that enzyme treatment improved hydrophilic properties. TCA-Soluble Peptides Content The highest content of TCA-soluble peptides was found in the chicken, followed by beef and squid muscle treated with 20% BE (w/w) respectively. The movement of concentration of BE is parallel with the movement of the TCA-soluble peptides as shown as in the page 6. Hydrolytic Patterns The myosin heavy chain (MHC) and acetin (AC) are the major proteins in all muscle foods. Apart from MHC and AC, a new protein band with high intensity, paramyosin(PM), was observed in the protein petterns of the squid muscle. The degradation of muscle protein also plays a major role in determining the tenderness and Water Holding Capacity of meat during the postmortem storage. Myofibrillar proteins such as troponin-T, titin and nebulin are considered to be indicators of the overall extent of postmortem proteolysis and predictors of meat tenderness.Result showed it could be determined that BE potentially has high proteolytic activity, making the meat more tender.
GRAPH CONCENTRATION vs TCA SOLUBLE PEPTIDES
TCA soluble peptides
Bromelain Extract Concentration (%, w/w)
Discussion
In this research study, it stated that bromelain act as enzyme that functioning in tenderizing meat protein structure, these experiment are the evidences of bromelain act as tenderizing agent. There are two type of properties that tested which is physically and chemically properties on beef, chicken and squid meat. For physically properties, it determined several test, first one is textural properties where shear force were taken place in this test, the result show the shear force value decrease when level concentration BE increased. The meat firmness causes of action proteolytic on myofibrillar protein. Second test are, water holding capacity and cooking yield, the result have been observed that there is significant reduction of WHC in all treated sample. The level of WHC may effect of pH level of meat, because pH value may responsible for overall reduction. There are three main factors involved in the shrinkage or swelling of the myofibrils, the onset of rigor mortis, the decline in pH and protein fragmentation. For cooking yield, the treated sample shown significantly different compared to control sample and marinated BE sample show reduction in cooking yield. This result indicate that the thermal treatment could remove water form treated. Reduction of cooking yield may cause by degradation of sarcoplasmic and myofibrillar proteins. Lastly are muscle microstructure test, based on the result, control sample shown the muscle fiber had well organized structure and closely bound each other compared to treated sample. This may due degradation of endomysial collagen and sarcolemma surrounding the muscle fibers also it show evident the degredation of muscle fiber occur where the fiber brokem and cell membrane degraded. For chemically properties experiment, first experiment have been carried out are pH and moisture content, there is significant different which is decrease in pH compare to control sample. The pH value in meat product is highly important because it has a major influence on other physic-chemical and quality properties such as WHC, tenderness and juiciness. Changes in pH also may due to the postmortem metabolism and application added substance to meat. For the Aw, bromelain powder extract on treated sample can absorb more water in fresh meat, automatically causing low Aw compared to control sample. The increased of BE concentration, the more reduction of Aw on meat, It is also probably due to the hygroscopic nature of BE powder when attached to the wet surface of the sample. The low moisture content of the treated samples is related to product acceptability for the consumer. Second experiment, TCA-soluble peptide, the hypothesis is the increase the concentration of BE, the highest TCA – soluble peptide in protein structure. From the result, the highest TCA – soluble content indicates greater muscle protein hydrolysis generated by proteolytic enzyme in BE. Last experiment is hydrolytic pattern, the myosin heavy chain (MHC) and actin (AC) are the major proteins in all muscle foods. A new protein band with high intensity, paramyosin (PM), beside (MHC) and (AC), observed in the protein patterns of the squid muscle. When comparing the treatment to the control, degradation of the AC and the MHC band was markedly degraded into lower molecular weight products. These enzymes have very broad specificities, and therefore unexpected break down the major muscle proteins where can give result on over – tenderization. During the postmortem storage, the degradation of muscle protein occur in meat which plays a major role in determining the tenderness and WHC. Indicators of the overall extent of postmortem proteolysis and predictors of meat tenderness are based on myofibrillar proteins. So it could be determined that BE potentially has high proteolytic activity, making the meat more tender.
GROUP MEMBERS
1. AHMAD SYAKIR BIN AHAMAD AZAHARI (F19B0010)2. RAJA ZAFRAN AIMAN BIN RAJA SALLEH (F19A0176) 3. MOHAMAD AIMAN SYAKIR BIN MOHAMAD SAITY (F19B0064) 4. MUHAMMAD MUAZ BIN MEFTAHUDDIN (F19B0265) 5. MUHAMMAD SYAHIRZAMIE BIN SHAHRUM (F19B0268)
Albert Einstein