EP0746211A1 - Produits alimentaires fabriques a partir de poisson renfermant l'enzyme protease, leurs procedes de fabrication et procedes pour inactiver l'enzyme protease dans le poisson - Google Patents

Produits alimentaires fabriques a partir de poisson renfermant l'enzyme protease, leurs procedes de fabrication et procedes pour inactiver l'enzyme protease dans le poisson

Info

Publication number
EP0746211A1
EP0746211A1 EP95944170A EP95944170A EP0746211A1 EP 0746211 A1 EP0746211 A1 EP 0746211A1 EP 95944170 A EP95944170 A EP 95944170A EP 95944170 A EP95944170 A EP 95944170A EP 0746211 A1 EP0746211 A1 EP 0746211A1
Authority
EP
European Patent Office
Prior art keywords
fish
merluccius
mixture
muscle
hake
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP95944170A
Other languages
German (de)
English (en)
Inventor
Gour S. Choudhury
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Alaska Fairbanks
Original Assignee
University of Alaska Fairbanks
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US08/361,824 external-priority patent/US5549920A/en
Priority claimed from US08/361,813 external-priority patent/US5518741A/en
Application filed by University of Alaska Fairbanks filed Critical University of Alaska Fairbanks
Publication of EP0746211A1 publication Critical patent/EP0746211A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/22—Working-up of proteins for foodstuffs by texturising
    • A23J3/26—Working-up of proteins for foodstuffs by texturising using extrusion or expansion
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/70—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/70—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor
    • A23L13/77—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor by mechanical treatment, e.g. kneading, rubbing or tumbling
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
    • A23L17/65—Addition of, or treatment with, microorganisms or enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
    • A23L17/70—Comminuted, e.g. emulsified, fish products; Processed products therefrom such as pastes, reformed or compressed products

Definitions

  • the present invention relates to food products made from protease enzyme containing fish, processes to make such food products, and processes to inactivate protease enzyme in fish, such as arrowtooth flounder.
  • the present invention also relates to a method for proteolytic degradation of fish that uses proteolytic enzyme (s) present in fish muscle to bring about heat-induced myofibrillar degradation, and to produce food products from the autolyzed or hydrolyzed mince.
  • proteolytic enzyme s
  • Alaska's flatfish resource is one of the largest in the world.
  • Arrowtooth flounder which constitutes about 65% of the flatfish biomass (excluding halibut) is presently unmarketable because of the presence of protease enzyme that degrades myosin. Enzymatic degradation of yosin during normal cooking leads to excessive softening of muscle resulting in an unacceptable paste-like texture of the cooked product. Lack of a suitable technology to inhibit/destroy protease in fish muscle, such as arrowtooth muscle, is preventing utilization of a vast untapped arrowtooth flounder resource off Alaska.
  • arrowtooth flounder may lead to development of a new fishery with marketing potential of about 386,180 metric tons a year. According to some fisherman of Kodiak, abundance of arrowtooth flounder may be preventing the resurgence of shrimp fisheries in the Gulf of Alaska. Utilization of arrowtooth flounder may help revive shrimp and other crustacean species.
  • the principal advantage of the present invention is the provision of a way to make use of the entire or almost the entire edible portions of fish containing protease enzyme, thus minimizing processing waste.
  • An additional advantage of the present invention is the development of food products from fish containing protease enzyme.
  • a further advantage of the present invention is the development of a process which will effectively inactivate protease enzyme in fish containing such enzymes in such a manner that the inactivation still permits products resulting from the process to be used as a food product.
  • the enzyme can be used in any application where proteolytic degradation is necessary or desired.
  • An additional goal is to develop a process by a suitable combination of proteolysis, drying and extrusion which will provide a commercially viable means for utilizing protease enzyme containing fish, such as arrowtooth flounder and other marine resources, including muscle from any fish, muscle from other animals (such as cattle, pigs, chicken, turkeys, and the like) and muscles recovered from animal by-products.
  • protease enzyme containing fish such as arrowtooth flounder and other marine resources, including muscle from any fish, muscle from other animals (such as cattle, pigs, chicken, turkeys, and the like) and muscles recovered from animal by-products.
  • the process of the present invention includes mixing protease enzyme containing fish meat or muscle with a starchy and/or proteinaceous material to form a mixture; and introducing this mixture into a screw extruder having barrel sections. At least one of the barrel sections is a reaction zone wherein the protease enzyme in the mixture is inactivated. At least one additional barrel section is located before the reaction zone to maintain the temperature of the mixture below the enzyme activation temperature of the fish meat or muscle.
  • the above objectives are also accomplished, in part, by a method to utilize protease enzyme present in fish muscle that includes distributing the enzyme throughout the entire fish or other animal muscle. Afterwards, the muscle is autolyzed and dried simultaneously or in sequence.
  • the autolyzed dried muscle is reduced to a powdered form.
  • a starchy and/or proteinaceous material is mixed with the powder to form a mixture which is subjected to high temperature processing such as elevated temperature extrusion processing to form a product which can be shaped in any form and subjected to further processing such as drying, baking, and flavoring to form food products.
  • the enzyme can be extracted from the muscle of fish containing protease enzyme and mixed with muscle from fish or any other animal (such as cattle, pigs, chicken, turkeys, and the like), or muscle by-products thereof.
  • the mixture is subjected to proteolysis and drying simultaneously or in sequence.
  • the present invention further relates to the food products formed by the above methods.
  • Figure 1 is a flow-diagram of one embodiment of the process of the present invention.
  • Figures 2a, 2b, and 2c show schematic diagrams of various screw configurations within a twin screw extruder for carrying out at least a part of the process of the present invention.
  • Figure 3 is a schematic diagram of a typical temperature profile within an extruder in accordance with the present invention.
  • Figures 4 and 5 are graphs showing the correlation between temperature and enzyme inactivation in arrowtooth flounder during extrusion.
  • Figures 6 and 7a are drawings illustrating different parts of a typical food extruder.
  • Figure 7b is a picture of a twin screw arrangement.
  • FIGS 8 and 9 are expanded perspective views depicting preferred die designs.
  • Figure 10 is a flow-diagram of one embodiment of the process of the present invention for fish muscle containing protease enzyme.
  • Figure 11 is a flow-diagram of one embodiment of the process of the present invention for fish muscle with or without protease enzyme.
  • Figure 12 is a flow-diagram of one embodiment of the process of the present invention for muscle from animals such as cattle, pigs, chicken, turkeys, and the like.
  • Figure 13 is a flow-diagram of one embodiment of the process of the present invention for muscle recovered from fish and other animal by-products.
  • Figure 14 is a graph showing the correlation between water activity and enzyme activity/microbial activity.
  • Fish muscle also referred to herein as fish meat from protease enzyme containing fish that can be processed by the present invention include protease enzyme containing flounders, hake, and menhaden.
  • protease enzyme containing flounders, hake, and menhaden examples include, but are not limited to, arrowtooth flounder (Atheresthes Stomias) , kamchatka flounder (Atheresthes Evermanni ) , yellowfin sole (Limanda Aspera) , Indian halibut (Psettodes Erumei) , Greenland halibut (Reinhardtius Hippoglossoides) , silver hake (Merluccius Bilinearis) , Chilean hake (Merluccius Gayi) , Argentine hake (Merluccius Hubbsi) , North Pacific hake (Merluccius Productus) , Benguela hake (Merluccius Polli
  • the protease enzyme for example, heat stable cysteine protease, present in arrowtooth muscle, causes rapid degradation of myosin heavy chain at elevated temperature.
  • the arrowtooth protease with sulphydryl group(s) at the active sites exhibits maximum activity at 55°C.
  • the enzyme is responsible for rapid proteolytic breakdown of myosin heavy chain and the degradation is complete within 20 minutes (Wasson et al. , J. Aq. Food Prof. Technol., Vol. 1, No. 3/4, p. 147 and p. 169, 1992; Greene and Babbitt, J. Food Sci., Vol. 55, No. 2, p. 579, 1990).
  • one method to inactivate protease enzyme in protease enzyme containing fish meat, such as arrowtooth flounder, and to make consumer-ready food products can be achieved in the following manner as shown in Figure 1.
  • the fish meat or muscle to be used in the process of the present invention can be obtained by removing the inedible portions of the fish, which include the head, guts, and backbone.
  • One way of preparing the fish meat to be used in the present invention is as follows.
  • a protease enzyme containing fish such as arrowtooth flounder, can be prepared for mincing by heading, gutting, and/or filleting the fish as those terms are understood to those skilled in the art.
  • fish preparation is known in the art, and, for instance, any fish filleting machine, such as a BAADER 175, can accomplish the filleting.
  • BAADER 175 can accomplish the filleting.
  • the fish is minced to remove the fish muscle from the skin and bones of the fish.
  • any conventional mincer can be used in the process of the present invention, for example, a belt and drum mechanical mincer (BAADER 697) .
  • the fish meat is preferably minced.
  • the fish meat is thoroughly mixed with at least one starchy and/or proteinaceous material, preferably at refrigerated temperatures (e.g., from about 4°C to about 10°C) .
  • starchy material include, but are not limited to, wheat flour, soy flour, rice flour, corn starch, corn meal, and the like.
  • proteinaceous material includes, but are not limited to, soy isolate, casein, whey protein, whey powder, wheat gluten, rice gluten, egg white powder, and the like.
  • the starchy and/or proteinaceous material absorbs the moisture from the fish muscle, acts as a binder with the fish muscle, and also increases the viscosity of the fish muscle after being mixed with the starchy and/or proteinaceous material, and forms a viscous paste.
  • any mixer can be used in the process of the present invention as long as it accomplishes this function.
  • a mixer is a RIBBON type mixer.
  • the mixture can be run at any normal speed (e.g., between 20 to 50 rpm) for approximately 1/2 hour to accomplish this mixing step.
  • the fish muscle used in the process of the present invention has a moisture content of about 70% to about 80% by weight of the fish muscle.
  • the starchy and/or proteinaceous material is preferably added in sufficient amounts to reduce this moisture content, for example to about 40% by weight of the fish muscle. Therefore, the starchy and/or proteinaceous material is preferably added in an amount of from about 5% to about 60% by weight of the fish muscle.
  • a powdery mix can be formed by using, for example, a HOBART mixer.
  • a dry feeder such as a K-TRON feeder, can then be used to feed the powdery mix to the extruder inlet.
  • the fish muscle and starchy and/or proteinaceous material can be a viscous paste or powdery mass.
  • the amount of starchy and/or proteinaceous ingredient generally used depends on the water absorption capacity of the latter. For example, soy flour has higher water absorption capacity than wheat flour.
  • the resulting mixture is a viscous paste or powdery mass depending upon the particular starchy and/or proteinaceous ingredient used, the amount of starchy and/or proteinaceous material used and the method of mixing. It is certainly within the bounds of the present invention to use one or more of the starchy and/or proteinaceous ingredients in the mixture.
  • a powdery mass is obtained, for example, when soy isolate is thoroughly mixed with minced fish muscle in an amount of about 20% by weight of the fish muscle.
  • Moisture in the fish mince may also be removed by processes such as squeezing before the mince is processed in the extruder. Another method of removing moisture from the fish mince can occur in the extruder itself by drawing a vacuum from a barrel section towards the end of the extruder immediately after an appropriate restriction created by a screw design. If needed, removal of moisture can be continued after the processed material exits from the extruder by further moisture removal means well known to those skilled in the art, e.g., by drying.
  • An extruder is a device which continuously processes a food material at high temperature and short time (HTST) .
  • Raw/processed material is fed into an elongated barrel by a feeder (51) , wherein the material is conveyed by one or more screws in the barrel assembly (52) and subjected to mixing, heat, and shear.
  • any means can be used to introduce the mixture into the extruder such as the use of a sanitary pump, for instance, a MOYNO pump.
  • the screw extruder permits a unique, high-temperature, short-time (HTST) processing of the mixture in order to inactivate the enzyme in the mixture.
  • HTST high-temperature, short-time
  • Protease in arrowtooth muscle exhibits minimum proteolytic activity below 40°C and has maximum activity between 55°C and 60°C.
  • the enzyme activity drops rapidly above 60°C. Less softening of arrowtooth muscle occurs with rapid heating suggesting inactivation of protease by denaturing the enzyme.
  • the sharp drop in enzyme activity above 60°C permits the use of HTST processing to inactivate this enzyme.
  • HTST processing of food offers better retention of nutrients and quality characteristics (color, flavor, texture) compared to conventional thermal processing such as canning.
  • D value decimal reduction time
  • z value represents the temperature range in °F for a 10:1 change in decimal reduction time.
  • HTST processing is based on the fact that heat-labile enzymes and microorganisms have much smaller D and z values than do nutrients and quality factors.
  • the rates of destruction that have small z values are highly temperature dependent; whereas rates with large z values are less influenced by temperature.
  • a given increase in temperature causes a larger increase in the rate of destruction of enzymes and microorganisms than in the rate of destruction of nutrients and quality factors. This enables HTST processing of foods with higher retention of nutrients and quality characteristics.
  • Increased water activity enhances the denaturation of enzyme protein.
  • enzymes In the presence of excess moisture, enzymes generally are much more temperature sensitive than microorganisms and are easily inactivated.
  • HTST treatment in a food extruder, preferably a twin screw extruder.
  • the process of the present invention includes direct feeding of protease containing minced fish muscle or the protease containing minced fish muscle preferably mixed with a starchy and/or proteinaceous material, and introducing this mince or mixture into a single screw or a multiple-screw extruder.
  • the extruder permits a unique high temperature short time (HTST) processing of the minced muscle or mixture in order to inactivate the enzyme in the mixture.
  • HTST high temperature short time
  • the mixture of fish muscle and starchy and/or proteinaceous material can be heated in a food extruder to very high barrel temperatures (e.g., 200°C-300°C) by a combination of mechanical and thermal energy with a very short residence time (about 1 to about 2 minutes) .
  • very high barrel temperatures e.g. 200°C-300°C
  • mechanical and thermal energy with a very short residence time (about 1 to about 2 minutes) .
  • the extruder is made up of a barrel, a screw (e.g., single, twin, or multiscrew) , a feeder, a die, a cutter, a drive-gear reducer and thrust bearing (54) , drive motor, and a heating and cooling arrangement with temperature control.
  • a screw e.g., single, twin, or multiscrew
  • a feeder e.g., a screw, a feeder, a die, a cutter, a drive-gear reducer and thrust bearing (54) , drive motor, and a heating and cooling arrangement with temperature control.
  • a food extruder has a drive, gear reducer, and thrust bearing (54) , a feed hopper (55) , a cooling water jacket (56) , thermocouples (57) , a barrel steam jacket (58) , a pressure transducer (59) , a die (60) , a discharge thermocouple (61) , a breaker plate (62) , a barrel with a hardened liner (63) , a metering section (64) , a compression section (65) , a feed section (66) , and a screw with increasing root diameter (67) .
  • a picture is shown of a twin screw arrangement that can be used in a food extruder.
  • the barrel in the extruder to be used in the present invention has several barrel sections and preferably has four, and more preferably five or six barrel sections as shown in Figures 2a, 2b, and 2c. At least one of these barrel sections is the main reaction zone where the HTST processing occurs.
  • the barrel sections prior to the main reaction zone permit a gradual increase in temperature of the fish muscle/starchy and/or proteinaceous material mixture prior to the mixture entering into the barrel section(s) which is the main reaction zone.
  • the temperature of the fish meat should not reach the maximum or optimal activation temperature of the protease enzyme in the fish meat (e.g., for arrowtooth flounder fish meat, a temperature of 40°-55°C) .
  • the first barrel section is where the mixture of fish meat and starchy and/or proteinaceous material is introduced into the extruder by means of a feeder (located above the first barrel section).
  • the temperature of the mixture in the first barrel section and the temperature of the fish meat in the optional barrel section(s), e.g., optional second barrel section, prior to the main reaction zone be below the activation temperature of the protease enzyme in the fish meat to be processed.
  • the temperature in the optional second barrel section will be higher than the first section and the gradual increase in temperature allows the mixture to be near, but not at, the optimal enzyme activation temperature for the fish meat in the mixture. By doing this, the HTST processing in the reaction zone is more effective since less time is spent reaching and exceeding the temperature for optimal enzyme activation.
  • the temperature of the mixture is below the enzyme activation temperature of protease enzyme containing fish.
  • the temperature of the mixture is below about 40°C, preferably about 5°C to about 10°C; and the mixture has a residence time in the first barrel section, of about 1 second to about 15 seconds, and more preferably about 2 seconds to about 10 seconds.
  • the temperature of the mixture is still below the enzyme activation temperature of the protease enzyme containing fish.
  • the temperature of the mixture is below about 40°C, preferably about 10°C to about 20°C; and the mixture has a residence time in the second barrel section of about 1 second to about 15 seconds, and more preferably about 2 seconds to about 10 seconds.
  • third barrel section i.e., third barrel section
  • a compressing section as shown in Figures 2a, 2b, and 2c.
  • the temperature of the mixture in this optional section is still below the enzyme activation temperature, which for arrowtooth flounder is at least about 40°C, preferably about 35°C to about 40°C; and the mixture has a residence time in this optional third barrel section of about 1 second to about 20 seconds, and more preferably about 2 seconds to about 15 seconds.
  • FIG. 2a A schematic of the barrel sections of a twin-screw extruder along with illustrative location of the mixing elements such as kneading and reverse screw elements is shown in Figures 2a, 2b, and 2c.
  • Screw configuration 1 (mild) of Figure 2a was built from the following screw elements: 50/50/2, 33.3/50/3, 25/50/3, and 16.6/50/2. These specifications are read as follows: pitch/length of screw element/number of screw elements. The total length of this combination of screw elements is 500 mm.
  • Protease enzyme in arrowtooth flounder for example, is completely inactivated in a Clextral BC-21 twin-screw extruder using screw configuration 1 (mild) , the barrel temperature profile shown in Figure 3, and a screw speed of 100 rpm.
  • the main reaction zone consisted of barrel sections 4 and 5 wherein the actual barrel temperatures were 90°C and 134°C. Enzyme inactivation under conditions described above is illustrative only. An inactivation profile can be changed by manipulating variables such as screw configuration, screw speed, and flow rates.
  • a wide choice of factors, such as screw configuration and screw speed for inactivating protease in fish are possible. These factors also affect the structure of the fish muscle due to their effects on energy inputs to the material. Any combination of the variables are acceptable as long as the protease is inactivated using the HTST processing.- In view of the present invention, manipulation of extrusion variables such as screw configuration, screw speed, and flow rate can be easily achieved to inactivate protease in a protease containing fish by one skilled in the art.
  • a typical product temperature in the die at exit of the extruder for inactivating protease in arrowtooth flounder fish meat using a Clextral BC-21 is about 100°C.
  • the "high temperature” in HTST is a temperature in which complete inactivation of the protease enzyme is achieved in the mixture. In the case of arrowtooth flounder, this is at least about 100°C, and preferably about 100°C to about 110°C.
  • the "short time" in HTST is about 5 seconds to about 30 seconds, and more preferably about 5 seconds to about 20 seconds.
  • HTST capability of food extruders destroys protease in protease containing fish, for example arrowtooth muscle, within a few seconds without sacrificing nutritional value and quality characteristics of muscle proteins.
  • the HTST treatment preferably occurs in the third barrel segment of the extruder (when there is at least a total of four barrel segments) and preferably occurs in the fourth barrel segment (when there is at least a total of five or six barrel segments) .
  • the main reaction zone there is an optional texturization zone.
  • at least one barrel section along with the die comprises this texturization zone.
  • the now inactivated fish mixture is subjected to barrel temperatures of at least about 200°C, preferably about 200°C - * > about 300°C, and more preferably about 200°C to about 250°C.
  • the texturization zone the fish mixture is converted to a fibrous texture that is easily formable into any desired shape. Accordingly, the texturization zone is used when it is intended to shape the fish mixture for food product use.
  • the barrel sections subsequent to the reaction zone can be used as a cooling section(s) (the fourth or fifth barrel sections) .
  • the enzyme inactivation using the high-temperature, short-time processing in the reaction zone can be achieved by manipulating screw configuration, temperature profile, die design, and residence time in the reaction zone within the extruder.
  • the screw configuration can manipulate mixing, residence time, and energy inputs to the feed material.
  • the screw profile typically consisted of elements of larger pitch of 50 mm in a conveying section (first and second segments of barrel) . This pitch is preferably gradually reduced to 33.3 mm, 25 mm, and 16.6 mm.
  • the total length of the screw in the screw extruder (made up of 25 mm elements and 50 mm elements) is 500 mm. Any length of screw is acceptable as long as the protease in the mixture of fish muscle and starchy and/or proteinaceous ingredients are inactivated.
  • a reverse pitch screw element and/or a kneading element can preferably be incorporated.
  • three preferred screw configurations in terms of mixing and shear are mild, intermediate, and severe as set forth in Figures 2a, 2b, and 2c, respectively.
  • FIGs 2a, 2b, 2c, and 3 a barrel having five sections is shown.
  • the screw configurations in Figures 2a, 2b, and 2c all have conveying sections.
  • the reaction zone has no mixing elements.
  • the reaction zone has a kneading element.
  • the reaction zone has a reverse pitch screw element and a kneading element.
  • twin-screw extruder is a Clextral BC-21.
  • a speed of about 50 rpm to about 400 rpm, and preferably about 100 rpm to about 300 rpm can be used with a residence time in the main reaction zone of about 5 to about 30 seconds.
  • the temperature range within the reaction zone must be sufficient to completely inactivate the protease enzyme in the fish being processed. For arrowtooth flounder, this temperature is approximately 100°C to about 110°C.
  • Figure 3 shows the typical temperature profile of the segments or barrel sections in the extruder.
  • the mixture of minced fish meat with starchy and/or proteinaceous material is slowly warmed up in the conveying sections but is not heated beyond the activation temperature (e.g., 40°C for arrowtooth flounder) until it is introduced into the reaction zone. Otherwise, the enzymes would be activated within the fish.
  • the temperature of the fish meat mixture is such that the protease enzyme is completely inactivated (e.g., at least about 100°C to about 110°C in order to inactivate the enzyme in the arrowtooth flounder) .
  • the average residence time in the reaction zone will be approximately about 5 to about 30 seconds which is sufficient to inactivate completely the enzyme.
  • the reaction zone can be located anywhere in the extruder as long as at least one conveying section is located before the reaction zone. Preferably, the reaction zone is located 100-200 mm from the end of the extruder. Screw speeds and flow rate can easily be varied by one skilled in the art to achieve the desired residence time in the reaction zone.
  • enzyme inactivation for arrowtooth flounder does not begin until a temperature of 40°C is obtained when the screw speed is 100 rpm; and 50°C when the screw speed is at a much faster rate, i.e., 300 rpm.
  • Substantially complete inactivation then occurs as can be seen in Figures 4 and 5 depending upon temperature and screw speed, but generally substantially complete inactivation of the fish meat mixture will occur when the temperature of the fish meat mixture is in the range of 70-110°C.
  • the other ingredients are added and mixed and the mixture is processed in subsequent segments to produce value-added consumer-ready food products.
  • enzyme inactivation and production of consumer-ready food products are combined in a single energy efficient, rapid, and continuous process.
  • the mixture exits from the die, which forms and shapes the mixture.
  • the die can be any design.
  • Figure 8 illustrates a texturizing die without temperature control while Figure 9 shows a variation of the design with facilities for temperature control. Flow of material is from C to A.
  • the die designs are compatible to different extruder types such as single screw and twin screw extruder.
  • the die hole inlet in the die of Figure 8 is circular and matches with the outlet hole from the die head of the extruder.
  • Side-View C-D shows the circular die hole inlet.
  • this die has a die opening (69) , an outer shell (70) , and a circular insert (71) .
  • the circular hole transits to an elliptical shape as shown in section G-H. Further transition of the shape takes place along the length of the insert to obtain the desired product shape at the die end.
  • a transition from an elliptical shape to a desired shape at the die end is a unique feature of the die design.
  • the final shape is shown as a rectangular die opening for the purposes of illustration.
  • the texturization die with temperature control can be attached directly to the extruder barrel. With respect to the texturizing die of Figure 9, this die also has a die opening (69) , an outer shell (72) , a circular shell (73) , a ring insert (74) , and a port for pressure or temperature sensor (75) . Figure 9 also shows where the fluid enters the die for purposes of cooling or heating (76) , and further shows where the fluid exits (79, 80) . Lastly, various welded joints (78) are shown.
  • the Side-View C-D illustrates a texturization die for a twin screw and Side-View A-B shows a rectangular die opening; however, in accordance with the present invention, it can be of any conventional shape or design.
  • Temperature control ( Figure 9, Sectional View E-F) is by heat transfer from or to a circulating fluid through the hollow die. This temperature control can effectively lower the temperature of the material after exiting the reaction zone and optionally the texturization zone. For instance, in the case of arrowtooth meat, in the texturization zone the barrel temperature is from about 200° to 300°C; while in the die, the material can be cooled to about 5°C to about 80°C.
  • the circular shell (see Figure 9) can also be hollow wherein heating coils can be inserted for supplying thermal energy, if necessary, to the material flowing through the texturizing die.
  • Length of both the texturizing dies shown in Figures 8 and 9 can be extended by similar attachments (sections) , and can further have features for varying the size and shape of the final product.
  • One way to have flexibility in this regard is to have replaceable inserts with different designs.
  • Both the hot and cold die can be used to texturize a fish product.
  • the mixture of fish muscle and starchy and/or proteinaceous material can be heated in the Clextral BC-21 extruder to very high temperatures (e.g., 300°C barrel temperature) by a combination of mechanical and thermal energy.
  • a multiple texture food product can be formed by attaching a co-extrusion die to the die end of the extruder. This permits, for instance, the fish food product to be covered by a different ingredient such as potatoes, flour, and the like.
  • the shaped mixture After exiting the die in the desired shape, the shaped mixture is cut into desired lengths by a conventional cutter and can then be subsequently smoked, refrigerated, frozen, and/or packaged.
  • additives such as spices, seasonings, colorants, nutrients, and anti-oxidants to enhance flavor, color, nutrition, and shelf life can be added during the process of the present invention.
  • additives include, but are not limited to coriander, ginger, onion powder, pepper, salt, and vinegar.
  • Flavoring, coloring, and other ingredients can be added to the feed material before entering the extruder, in the extruder, or after the material exits from the extruder.
  • the process of the present invention can be modified by raising the temperatures in all or part of the extruder barrels above 200°C and feeding a mixture of fish mince and starchy and/or proteinaceous ingredient, and forming and shaping the extrudate, preferably through a die having a design illustrated in Figures 8 and 9.
  • the process uses any combination of proteolysis, drying, and extrusion to fabricate value-added food products from arrowtooth flounder and other fish having similar proteolytic enzymes, such as those named earlier.
  • the protease enzyme containing fish to be used in the process of the present invention can be obtained by removing the inedible portions of the fish, which include the head, viscera, and backbone.
  • One way of preparing the fish to be used in the present invention is as follows.
  • a protease enzyme containing fish such an arrowtooth flounder, can be prepared for mincing by heading, gutting, and/or filleting the fish as described earlier. Once this initial preparation is completed, the fish is preferably minced to remove the fish muscle from the skin and bones of the fish.
  • the fish muscle obtained is stirred in a mixer to facilitate uniform distribution of the muscle protease throughout the fish.
  • the speed setting of the mixer and the length of time in the mixer are dependent on the type of mixer, and the amount of fish being mixed.
  • the mixer can be any conventional mixer as long as substantially uniform distribution of the enzyme throughout the meat is accomplished. For instance, in a RIBBON mixer, a preferred speed is from about 20 rpm to about 5 rpm, for about 10 minutes to about 40 minutes.
  • the mixed fish muscle is then subjected to proteolysis and drying.
  • the proteolytic degradation of muscle proteins and drying of the fish muscle can be carried out simultaneously or in sequence.
  • the heat-induced myofibrillar degradation is completed during the initial phase of drying.
  • gradual loss of enzyme activity will occur.
  • the removal of water during drying decreases water activity of the fish muscle.
  • the decrease in water activity is accompanied by gradual loss of enzyme activity and suppression of microbial activity (Schwimmer, Food Techol., Vol. 34, No. 5, p. 64, 1980; Troller, Food Technol., Vol. 34, No. 5, p. 76, 1980) .
  • the muscle can be first hydrolyzed and then dried.
  • the hydrolysis can be carried out in a reactor vessel connected to a food extruder or in the reactor vessel alone.
  • the fish mince is brought to the optimum enzyme activation temperature in a food extruder and dropped in the reactor vessel, which is maintained at the optimum temperature for the enzyme action.
  • the mince is then stirred in the reactor vessel until the desired degree of proteolysis is achieved.
  • the mince can be added to a reactor vessel, brought to the optimum enzyme activation temperature and stirred at that temperature until the desired degree of proteolysis is achieved.
  • the hydrolyzed muscle proteins can either be dried or fractionated before drying. The dried muscle or fractions thereof can then be used for the production of food products ( Figures 10-13) .
  • any conventional dryer can be used as long as the dryer is capable of reducing the fish muscle moisture content to about 10%.
  • drying is carried out for from about 10 minutes to about 4 hours at a temperature of from about 55°C to about 60°C and then for about 7 hours at a temperature range of from about 70°C to about 95°C.
  • the temperature of the arrowtooth flounder muscle be maintained at 55°-65°C for the initial drying phase that can range from about 1 minute to about 4 hours, and preferably about 10 minutes to about 4 hours depending on the degree of proteolysis desired.
  • the temperature during drying (second phase) will depend on the type of dryer used.
  • a preferable temperature range for the second phase can be from about 70°C to about 110°C.
  • the drying time will depend on the type of dryer used. For example, 60 pounds of fish muscle can be dried (second phase drying) in a pilot scale tray dryer in about 7-8 hours. For fractions containing soluble proteins, a spray dryer can alternatively be used.
  • the dried autolyzed fish meat is then reduced to powder, mixed with starchy and/or proteinaceous ingredients, and subjected to extrusion processing.
  • Reducing the dried fish muscle into powder can be accomplished through conventional means known to those skilled in the art, including the use of Hammer Mill or Urschel Comitrol size reduction equipment. It is preferred that the dried fish meat be reduced to a size of about 10 to about 400 mesh, more preferably about 10 to about 100 mesh, and most preferably about 40 mesh.
  • starchy (or starch) ingredients or materials include, but are not limited to, rice flour, wheat flour, corn starch, corn meal, soy flour, and the like.
  • proteinaceous materials or ingredients include, but are not limited to, soy isolate, casein, whey protein, whey powder, wheat gluten, rice gluten, egg white powder, and the like. Although there is no intention to limit the amount of starchy and/or proteinaceous material to be added to the dried and powdered fish muscle, it is preferred that the resulting mixture have the ratio of about 5% to about 60% by weight fish muscle to about 40% to about 95% by weight starchy and/or proteinaceous material; more preferably about 5-30% fish muscle to about 70-95% starchy and/or proteinaceous material; most preferably about 5-20% fish muscle to about 80-95% starchy and/or proteinaceous material.
  • the dried and powdered fish should be sufficiently mixed with the starchy and/or proteinaceous material and water so that a uniform mixture is obtained.
  • the uniform mixture is then introduced into an extruder, preferably a twin screw extruder.
  • the high temperature extrusion is at a barrel temperature of from about 130°C to about 190°C, preferably about 150°C to about 165°C. It is also preferable that a reverse screw element and or kneading element be part of the screw configuration.
  • the mixture exits the extruder and enters a die opening/configuration which will form and shape the mixture.
  • High temperature extrusion processing will destroy any residual enzyme activity and eliminate or reduce the microbial population.
  • extrudate shaped by a suitable die will then be cut into pieces of desired length and subjected to post-extrusion processing such as drying, baking, flavoring, and packaging to produce value-added nutritive savory food products.
  • the protein mix obtained after proteolysis and the fractions obtained after proteolysis and fractionation can also be mixed with starchy and/or proteinaceous ingredients and extruded at a barrel temperature of about 200°C to about 300°C to form a texturized food product.
  • the use of proteolytic enzyme activity present in fish muscle can be used to hydrolize muscle from any other fish or animal, including by-products thereof.
  • enzyme containing fish muscle can be blended with water and tissuemized (i.e., the tissue is crushed into very small particles) . Then the mixture can be centrifuged to form a crude enzyme extract. This extract can then be mixed with any fish or other animal muscle.
  • the muscle can be partially broken down (i.e., tenderized) or substantially or completely broken down (i.e. about 10% to about 100% proteolytic degradation of the fish or other animal muscle) .
  • the substantial or complete degradation process can be enhanced if the fish or other animal muscle to be hydrolized is minced prior to being subjected to proteolysis.
  • the enzyme extract can be used to tenderize fish or other animal muscle.
  • the enzyme extract can be used to cause entire high molecular protein polymer breakdown (proteolytic degradation) in fish or other animal muscle.
  • the fish or animal muscle can be dried and/or mixed with a starchy or proteinaceous material and extruded as described above to make food products.
  • Protease in arrowtooth mince was inactivated under the following conditions.
  • a Clextral BC-21 twin-screw extruder was used having a screw configuration consisting of only conveying elements (no mixing elements) similar to that shown in Figure 2a.
  • Screw configuration from feed to die end was: 50/50/2, 33.3/50/3, 25/50/3, and 16.6/50/2. These specifications are read as follows: pitch/length of screw element/number of screw elements. The total length of this combination of screw elements was 500 mm.
  • the barrel temperature profile in the five barrel sections from feed to die was 0, 0, 47, lio, 140°C, respectively.
  • Arrowtooth mince was mixed with 1% (by total weight of mixture) soy isolate and was fed to the extruder using a MOYNO pump.
  • the screw speed, throughput, and product temperature at the die were 300 rpm, 18 kg/hr, and 99°C, respectively.
  • the die was circular with a diameter of 10 mm. Protease enzyme in the arrowtooth mince was completely inactivated.
  • a dual textured or multiple-textured product can be formed using minced fish.
  • a dual textured product was produced by first inactivating protease enzyme in arrowtooth using the extruder and procedures in Example 1. Then, the extruded arrowtooth meat was fed into the inner core of a co-extrusion die using a MOYNO pump.
  • the outside coating was rice flour which was fed as a powder from a hopper to the extruder.
  • the screw configuration from feed to die end was*. 50/50/2, 33.3/50/3, 25/50/3, and 16.6/50/2.
  • the specifications are read as follows: pitch/length of screw element/number of screw elements.
  • the total length of this combination of screw elements was 500 mm.
  • the barrel temperature profile in the five barrel sections from feed to die was 20, 40, 60, 80, 100°C, respectively.
  • the screw speed, throughput, and product temperature at the die were 100 rpm, 12 kg/hr, and 70°C, respectively.
  • a well formed dual texture product was formed. This product could then be further coated using a dipping or spraying process to form a multiple-textured product, if desired.
  • a textured product can be formed using a hot die or a cold die by manipulating independent process variables such as screw configuration, die design and temperature profile.
  • a mix of arrowtooth flounder and 25% (by total weight of mixture) wheat flour was processed in a Clextral BC-21 twin-screw extruder using a mild screw configura ion (no mixing elements) similar to Figure 2a.
  • Screw configuration from feed to die end was: 50/50/2, 33.3/50/3, 25/50/3, 25/25/1, 16.6/50/3, and 16.6/25/1. These specifications are read as follows: pitch/length of screw element/ number of screw elements. The total length of the barrel was 600 mm with 6 barrel sections of 100 mm each.
  • Flow rate was 18 kg/hr and screw speed 200 rpm.
  • Set barrel temperatures were 200°C in the first barrel section and 265°C in the other 5 barrel sections.
  • the actual barrel temperature was lower than the set barrel temperature, and the material temperature in the extruder was lower than the actual barrel temperature.
  • the actual barrel temperatures were 43°C and 265°C, respectively, and the material temperature inside the sixth barrel section was 198°C.
  • a slit die (15 mm x 2 mm) was used, and the material temperature was 144°C in the die.
  • a well textured fibrous structure was formed in a continuous strip under the above conditions.
  • a textured product can also be made from minced fish which do not have protease problems.
  • a mix of 75% minced salmon and 25% (by total weight of mixture) wheat flour was extruded in a Clextral BC-21.
  • Screw configuration from feed to die end was: 50/50/2, 33.3/50/3, 25/50/2, KB/5/5, LH16.6/25/1, 25/25/1, 16.6/25/1, KB/90/5/5, LH16.6/25/1, and 16.6/50/2.
  • the total length of the barrel was 600 mm with 6 barrel sections of 100 mm each.
  • Set barrel temperature are 220°C throughout.
  • the actual barrel temperatures from feed to die end were 39, 104, 172, 216, 220, 220°C, respectively.
  • a rectangular die section 35 mm x 5 mm produced a well textured product at 200°C barrel temperature. Flow rate of the mix was 23 kg/hr. A well structured fibrous product was formed.
  • a textured product can be made from deep water fish species such as giant grenadier which has very high moisture content (approximately 90%) .
  • a mix of 60% giant grenadier mince and 40% (by total weight of mixture) wheat flour was extruded in a Clextral BC-21.
  • Screw configuration from feed to die end was: 50/50/2, 33.3/50/3, 25/50/3, 25/25/1, 16.6/50/3, and 16.6/25/1.
  • pitch/length of screw element/ number of screw elements was 600 mm with 6 barrel sections of 100 mm each.
  • the set barrel temperature was 220°C throughout.
  • the actual barrel temperatures from feed to die end in six sections were 39, 100, 166, 202, 218, and 220°C, respectively.
  • the product temperature in the die was 139°C.
  • Flow rate of the mix was 18 kg/hr and screw speed was 100 rpm.
  • a rectangular slit die (15 mm x 2 mm) produced a well textured product at barrel temperature of 220°C.
  • Minced arrowtooth flounder was dried in an Enviro-Pack dryer/smoker at a wet bulb temperature of 55°C and a relative humidity of 70%.
  • the autolyzed dried mince was then ground in an Urschel Comitrol and sieved to obtain a 40 mesh powder.
  • a mixture of hydrolyzed arrowtooth powder (15% fish solids) and rice flour (85% rice solids) was prepared at a moisture content of 10%.
  • the mixture was extruded in a twin-screw extruder (Clextral BC 21) at 160°C through a 5 mm diameter circular die.
  • the set barrel temperatures from feed to die end were 0, 0, 0, 100, 160, 160°C, respectively.
  • the actual barrel temperatures from feed to die end were 14, 14, 15, 97, 157, 160°C, respectively.
  • Feed rate and screw speed were at 12 kg/hr and 400 rpm respectively.
  • the extruded product was very good in appearance (slightly brownish tinge) and had excellent expansion characteristics (expansion ration of 16.63).
  • the soluble protein concentration over a period of one hour increased linearly by 13 ⁇ g/ ⁇ l, 12.5 ⁇ g/ ⁇ l and 25 ⁇ g/ ⁇ l for Pollack mince, frame and skin respectively. These values were obtained after correcting for native soluble proteins, mechanical liberation of peptides by stirring and autoproteolysis.
  • the frames were clean and free of adhering muscle tissue.
  • the apparent breakdown of skin and intervertebral cartilaginous tissues indicate the affinity of the enzyme for these substrates. This affinity indicated that the enzyme has ability to tenderize and reduce toughness of meat.
  • arrowtooth protease The ability of arrowtooth protease to hydrolyze muscle of domestic meat animals was tested by incubating beef and pork with protease at optimal condition for enzyme action.
  • Arrowtooth mince and water were blended in a 2:3 ratio, tissuemized in a Tekmer Tissuemizer (Type SDI-1810) for 6 minutes, and centrifuged at 10,000 g for 10 minutes in a refrigerated (4-5°C) centrifuge (Sorvall Dupont, Model RC5C) .
  • the crude enzyme extract (supernatant) was mixed with cubes of beef and pork in conical flasks and maintained at 55°C in a water bath. Controls were subjected to same reaction conditions with water substituted for the crude enzyme extrac .
  • the process of the present invention will utilize a vast supply of valuable proteins and produce a volume of consumable products.
  • the process of the present invention will also produce nutritive savory products that will compete in the snack food market and will provide access to diverse national and international markets.

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Abstract

Procédé destiné à inactiver l'enzyme protéase dans un poisson, tel que la plie à grande bouche, renfermant dans une partie de ses organes cette enzyme. Au cours du traitement, le poisson est coupé en morceaux puis mélangé à une substance amylacée et/ou protéinique pour obtenir un mélange qui est ensuite soumis à un traitement court, à haute température, dans une extrudeuse comportant une zone de réaction. La présente invention se rapporte également à un procédé destiné à la dégradation protéolytique des muscles du poisson renfermant l'enzyme protéase, procédé consistant à répartir uniformément l'enzyme dans tout le muscle du poisson, puis à sécher celui-ci. La présente invention se rapporte encore à un procédé de fabrication d'un produit alimentaire qui consiste à réduire le muscle hydrolysé du poisson pour le transformer en poudre, puis à mélanger la poudre obtenue avec une substance amylacée et/ou protéinique afin d'obtenir un mélange qui est ensuite soumis à un processus d'extrusion à haute température pour obtenir un produit alimentaire désiré. La présente invention se rapporte aussi à des procédés visant à attendrir le muscle de l'animal ou à provoquer une dégradation protéolytique substantielle ou totale de celui-ci en appliquant sur le muscle l'enzyme protéase prélevée dans le poisson. L'invention se rapporte aux produits alimentaires obtenus par ces procédés.
EP95944170A 1994-12-22 1995-12-21 Produits alimentaires fabriques a partir de poisson renfermant l'enzyme protease, leurs procedes de fabrication et procedes pour inactiver l'enzyme protease dans le poisson Withdrawn EP0746211A1 (fr)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US08/361,824 US5549920A (en) 1994-12-22 1994-12-22 Extrusion inactivation of protease enzyme in fish and fish food products
US08/361,813 US5518741A (en) 1994-12-22 1994-12-22 Product and process for the utilization of enzyme and muscle from fish containing proteolytic enzymes
US361813 1994-12-22
US361824 1994-12-22
PCT/US1995/016641 WO1996019120A1 (fr) 1994-12-22 1995-12-21 Produits alimentaires fabriques a partir de poisson renfermant l'enzyme protease, leurs procedes de fabrication et procedes pour inactiver l'enzyme protease dans le poisson

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DE10054516A1 (de) * 2000-11-03 2002-05-16 Henkel Kgaa Extrudiertes Proteinhydrolysat, Verfahren zu dessen Herstellung und dessen Verwendung
WO2003022076A1 (fr) * 2001-09-10 2003-03-20 Nippon Suisan Kaisha, Ltd. Produits a base de chair de poisson de fond obtenus a partir de chair de poisson contenant une protease
US8293297B2 (en) * 2007-04-05 2012-10-23 Solae, Llc Colored structured protein products
RU2013150130A (ru) 2011-04-12 2015-05-20 Геа Фуд Сольюшнс Бакел Б.В. Способ производства мясоподобного продукта
US9820504B2 (en) 2013-03-08 2017-11-21 Axiom Foods, Inc. Rice protein supplement and methods of use thereof
CN108271917A (zh) 2013-03-08 2018-07-13 艾斯姆食品公司 大米蛋白补充剂
EP3223625B1 (fr) * 2014-11-24 2022-04-27 Société des Produits Nestlé S.A. Aliments à mâcher pour animaux comestibles à masse volumique faible et leurs procédés de fabrication
WO2018209131A1 (fr) 2017-05-12 2018-11-15 Axiom Foods, Inc. Produits de riz et systèmes et procédés de fabrication de ceux-ci
CN109773847B (zh) * 2019-03-08 2023-10-24 荣成泰祥食品股份有限公司 一种鱼豆腐切块装置及鱼豆腐的生产方法

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JPS60188045A (ja) * 1984-03-08 1985-09-25 リサーチ・コーポレイシヨン 保存安定性魚肉ベース製品
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JPH0775529B2 (ja) * 1986-02-17 1995-08-16 株式会社あじかん 魚介肉すり身の組織化物の製造法
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AR000519A1 (es) 1997-07-10
CA2183057A1 (fr) 1996-06-27

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