|l||||||||||||||||||||||||l||||||||l||||||l||||||||l|||||||||||l||||l|l||||
`
`US00’}'7637l7Bl
`
`p
`
`O
`02; United States Patent
`Jaczynski
`
`(S4) CONTINUOUS PROTEIN AND LIPID
`RECOVERY FRUNI FOOD ANINIAL
`PROCESSING nYpR()[)U(_‘Ts
`
`(75)
`
`Inventor:
`
`Jacek Jaczynski. Morgantown. WV’
`(US)
`
`_
`_
`I
`I
`I
`_
`[73] Assigiiee: West Virginia University Research
`C011). 0fWCSt Virginia UI'Ii\’Cl‘Sit}r‘.
`Morgantown, VVV (US)
`
`( “‘ ) Notice:
`
`Subject to any disclainicr. the term ofiiijs
`pmcm is extended 0,. adjusted under 35
`USE‘ 154w) by 93 days_
`(21) Appl. No; 111369.331
`
`(10) Patent No.:
`(45) Date of Patent:
`
`US 7,763,717 B1
`Jul. 27, 2010
`
`6.887.508 132*
`T.(.|Ol.634 B2 “
`2t)03»'(J124239 AI
`20049006755] Al
`
`5.-"20C-5 Huang
`232096 Browne
`‘P2003 Keileher
`452004 Hultin et a].
`
`4269573
`4263329
`
`"F2005 Kenehe’
`2DC'5"'0233U60 "M
`OTHER PUBLICATIONS
`Study, l. M. ( I 999) Protein Denatuietion in Foam. .T.Col|o1dlnterf:1ce
`39;“ mi 215_ No_ 3_ pp_ 333-332,-+
`Mleko el al. (1997) Interactions of kappa. -ca.rr:1gee11a.n with whey
`proteins in gels formed at diflcicnt pH, Food Res. International. vol.
`30. No. 6. pp. 427-433.”
`1\:Ii1Woevskii er a1. (29040 A homoscnizt-=r - A new In‘--2 0f‘tiulVcI'iZe1'-
`Cl1€Il'I.‘PCi[DiCL|m Eng. vol. 40, l\to_s.|l 1|-12, pp. 1551-654.
`1
`Eiillirigocct &(Iil.9(?l29)7Eg }:'t$1?;:1;2l11 toi bitiltery pioducts. J. Am. Oil
`"‘ cited by examiner
`
`Mar‘ 71
`Reiated U.S_ Application Data
`(60) Provisional application No. 60!659,685. filed on Mar.
`8. 2005.
`
`£‘)?'l'-iifflifil-?'_l’f'E.':\‘.'al'."l.?'i7-38?’
`.AS.lla.l1dII_I[ii:-)C5EIi
`5.51.5’ CH1
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`0.?’ FErm—Mary-Jacq l-Iolroyd;
`o
`ston
`0 to
`Y
`
`(57)
`
`ABSTRACT
`
`(51)
`
`Im_ CL
`A231 M90
`(2005131)
`A (“K 38/00
`(200501)
`{52) u.s. Cl.
`s:+tir4i2; 5307355; 5307205;
`420,652
`,
`,
`,
`gage]: °¥::l:'is(::fig£"f?)];cS:;lrT:te' Nme
`pp
`P
`'
`Rgferences cm-d
`U.S. PATENT DOCUMENTS
`
`(58)
`(55)
`
`A proce-sis and system for recovering protein and lipid from
`food animal byproducts, and the products thereof, involves
`hoinogcnizing enin-ml byproducts with water to lorm a homo-
`3°“a‘*’- 5°‘“b'1m“9>"‘” “°“1°3"~“‘a‘°bY_*‘1'“5“”9>"1"I?1i] °f
`the h0IIt'l0g(::laI[E3 to
`a Edrst pl-1 adiusted D0l:l'tp0Sl:It0}lI:,
`separating t e irst p - a gust
`composition orniing a 1g 1
`ft"actioii_oontaiii_iiig lipids (oil). a inediiitn fraction‘containing
`protein in solution, and a heavy fraction containing fat~free
`impurities, separation by first centrifugatioti. adj usting the pH
`of the medium: fraction to about the iS(‘ieieCI1'iC point of the
`proteins thereby precipitating the inecliutn fraction forming a
`second pll adjusted composition, and separating the second
`pH adjusted composition forming a light t'i-action containing
`water and E1 heavy fmctioii contajiting precipitated pmtciiis.
`The water may then be recycled and used in the hoi1.1ogt:ii.i-
`zzition of furtlier byproducts.
`
`..
`
`.
`4263646
`
`5.171.592 A “‘
`
`5.384.]-49 A “
`6.005.073 A
`6.86.959 A
`6.288.2lIS Bl
`(i.45l.9'i’5 Bl *
`
`12-"1992
`l2"1994
`1: I995
`121999
`IUQ000
`.
`9.’200l
`92003 I-lultin eta].
`
`.--2103639 5.3?2.723 A "‘
`
`530.350
`
`32 Claims, 10 Drawing Sheets
`
`EXHIBIT
`
`5+
`
`AKBM 1087
`
`AKBM 1087
`
`1
`
`

`
`U.S. Patent
`
`Jul. 27, 2010
`
`Sheet 1 of 10
`
`US 7,763,717 B1
`
`HOMOGENATION
`
`FIRST PH ADJUSTMENT
`
`I6
`
`FIRST SEPARATION
`
`
`
`
`HEAVY FRACTION,
`FAT-FREE IMPURITIES
`
`
`
`MEDIUM FRACTION
`PROTEIN SOLUTION
`
`
`LIGHT FRACTION OIL
`
`24
`
`SECOND PH ADJUSTMENT
`
`
`
`SECOND SEPARATION
`
`HEAVY FRACTION:
`PRECIPITATE
`PROTEINS
`
`LIGHT FRACTION
`WATER
`
`
`
`
`32
`
`Fig. 1
`
`34
`
`2
`
`

`
`U.S. Patent
`
`Jul. 27, 2010
`
`Sheet 2 of 10
`
`XL 7,763,717 B1
`
`
`
`
`FEEDING SUBSTRATE
`
`PROCESSING
`
` FAT-FREE
`IMPURITIES
`
`HARVESTING PROTEIN PRODUCTS
`
`47
`
`'
`
`FAT—FREE
`IMPURITIES
`
`
`
`Fig. 2
`
`FEEDING
`SUBSTRATE
`
`
`
`PROCESSING
`
`
`
`40
`
`HARV ESFIN G
`
`PRODUCTS
`
`I2
`
`Fig. 3
`
`3
`
`

`
`U.S. Patent
`
`Jul. 27, 2010
`
`Sheet 3 of 10
`
`US 7,763,717 B1
`
`PROTEIN
`PRODUCTS
`
`SECOND CENTRIFUGE
`
`
`
`FIRST BIO-REACTOR
` FIRST CENTRIFUGE
`52
` OIL
`
`
`
`SECOND BIO-REACTOR
`
`50
`
`45
`
`47
`
`
`
`FAT-FREE
`I MPU RITIES
`
`Fig. 4
`
`4
`
`

`
`U.S. Patent
`
`Jul. 27, 2010
`
`Sheet 4 of 10
`
`US 7,763,717 B1
`
`HOMOGENJZATION
`BY-PRODUCT: WATER
`
`1:6 (WT:WT)
`
`
`
`
`
`
`
`
`SULUBILIZATION
`AT PH= 12.0
`
`1ST SEPARATION
`10,000 X G FOR 10 MIN AT 4C
`
`FISH LIPIDS
`
`22‘
`
`I8‘
`
`
`
`
`
`IMPURITIES (BONES,
`PROTEIN SOLUTION
`SKIN, SCALE, INSOLUBLE
`
`
`PROTEINS, ITC.)
`
`
`25'
`PRECIPITAHON
`AT PH-5.5
`
`
`PROTEINS TRANSFERRED
`TO JAR TESTER WITH SIX JARS
`
`58
`
`
`
`
` 'NIONIC FLOCCOILULA
`CATIONIC FLOCCULANT
`NONIONIC FLOCCULANT
`
`5
`
`

`
`U.S. Patent
`
`Jul. 27, 2010
`
`Sheet 5 of 10
`
`Us 7,763,717 B1
`
`IEIIIIIIIKl.a'i'a.Z.i.i'Hl2'n'i'K'
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`
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`
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`
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`
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`
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`
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`
`0505055?_fl.J_.J7_
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`
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`
`30
`
`40
`TIME (MIN)
`Fig. 7
`
`6
`
`
`
`
`
`

`
`U.S. Patent
`
`Jul. 27, 2010
`
`Sheet 6 of 10
`
`US 7,763,717 B1
`
`E CONTROL [H25 MG/L E so MG/L
`
`250 MG/L '-5:”
`
`:L.,GMmWH...GMBU5
`
`2 000
`
`
`
` -IE:2.3Emzuo._§.Eo
`
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`
`
`
`50
`
`60
`
`70
`
`0S
`
`40
`30
`TIME (MIN)
`Fig. 8
`
`2.5/5.0
`
`2.0X6.0
`
`125155
`
`‘l2.5l5.0
`
`1 2.05.5
`
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`
`1 3.0;’5.5
`
`SOLU BILIZATIO NIPRECI PITATION
`PROTEIN RECOVERY YIELDS
`
`Fig. 9
`
`7
`
`
`
`
`
`

`
`U.S. Patent
`
`Jul. 27, 2010
`
`Sheet 7 of 10
`
`Us 7,763,717 B1
`
`2 5 A
`
`/00
`
`1.5 '
`
`—-
`,.,
`E;
`2 2° 7
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`
`PROTEIN SOLUBILITY AND PRECIPITATION PROFILES
`
`Fig. 10
`
`9 <
`
`9
`
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`
`«b-O
`
`INJD ELECTRIC
`
`MODULES(KPA) 8‘:5’
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`1 O
`20
`30
`40
`50
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`
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`
`80
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`
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`
`Fig. 11
`
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`
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`—A— SARCOPLASMIC
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`
`

`
`
`
`U.S. Patent
`
`Jul. 27, 2010
`
`Sheet 8 of 10
`
`US 7,763,717 B1
`
`
`
`KRAMER
`
`PUNCH HARDNESS
`
`SHEAMGIGJ
`
`(G)
`
`(G)
`
`
`
`COHESIVENESS GUMINESS CHEWINESS
`
`1159.26
`
`1018.45
`
`1907.75
`
`1583.15
`
`
`
`
`
`
`
`8-69
`
`
` TROUT PROTEIN GELS
`
`
`
`E 7
`
`% ASH
`
`IKRILLTAILMEAT I 11.09
`WHOLE KRILL
`17.36
`2.0 I
`5.93
`$2.5
`4.32
`.0
`4.01
`12.0
`4.88
`
`I
`
`I
`
`12.5
`13.0
`
`5.71
`5.74
`
`I
`
`I
`
`I;
`2 3
`E E
`O E
`Di :1
`9- 5
`tn
`
`Fig. 13
`
`%ASH
`
` BONELESS SKINLESS TROUT FlLLE|' 5.54
`TROUT FRAMES (BY PRODUCTS]
`13.91
`RECOVERED FAT-FREE IMPURITIES
`41.10
`
`I
`
`PROTEINS
`
`SOLUBILTZED
`
`2.5I 2.14
`3.0
`1.61
`
`I
`
`12.0
`12.5
`13.0
`
`0.88
`1.37
`2.14
`
`Fig. 14
`
`9
`
`

`
`U.S. Patent
`
`Jul. 27, 2010
`
`Sheet 9 of 10
`
`US 7,763,717 B1
`
`ESSENTIAL AMINO ACIDS
`
`THR
`
`TROUT 1.8
`FRAMES
`
`VAL
`
`2.2
`
`MET
`
`1.4
`
`ILE
`
`1.8
`
`LEU
`
`3.1
`
`PHE
`
`1.6
`
`HIS
`
`1.2
`
`LY5
`
`3.5
`
`TRP
`
`TOTAL AVERAGE
`
`0.5
`
`17.2
`
`17.2
`
`2.0
`
`3.7
`
`4.6
`
`2.6
`
`3.9
`
`6.6
`
`3.4
`
`2.1
`
`7.4
`
`1.0
`
`35.3
`
`2.5
`2:
`3.4
`4.3
`2.2
`3.5
`5.0
`3.1
`1.9
`5.7
`0.9
`32.3
`EB 3.0
`3.7
`4.7
`2.5
`4.0
`5.5
`3.4
`2.1
`7.3
`0.9
`35.2
`E‘? 12.0 '3.s '5.o '25 '4.2 '5.9 ' 3.5' 23' 7.5' 1.1' 312'
`E981
`12.5
`3.9
`4.9
`2.5
`4.1
`5.9
`3.5
`2.2
`7.5
`1.1
`35.9
`
`34.3
`
`37.4
`
`'
`
`7.8
`
`1.2
`
`38.2
`
`13.0
`
`4.1
`
`5.1
`
`2.6
`
`4.3
`
`7.1
`
`3.7
`
`2.3
`
`WHOLE 2.2
`KRILL
`
`2.6
`
`1.5
`
`2.5
`
`4.0
`
`2.2
`
`1.1
`
`4.4
`
`0.7
`
`21.2
`
`21.2
`
`2.0
`
`4.8
`
`6.0
`
`2.9
`
`5.5
`
`9.0
`
`5.0
`
`4.9
`
`2.6
`
`2.5
`
`9.2
`
`9.2
`
`1.5
`
`1.6
`
`46.6
`
`46.3
`
`47.0
`
`SOLUBILIZEDATpH
`
`
`
`KRILLPROTEINS
`
`2.5
`
`3.0
`
`12.0
`
`12.5
`
`13.0
`
`4.5
`
`4.8
`
`4.6
`
`4.5
`
`4.4
`
`SOYBEAN 3.9
`
`FNB
`
`3.5
`
`5.8
`
`5.9
`
`5.8
`
`5.6
`
`5.5
`
`4.6
`
`4.8
`
`3.2
`
`3.3
`
`3.4
`
`3.2
`
`3.1
`
`1.1
`
`2.6
`
`5.7
`
`5.9
`
`5.7
`
`5.5
`
`5.5
`
`4.6
`
`4.
`
`8.9
`
`9.2
`
`8.8
`
`8.6
`
`8.4
`
`7.8
`
`7.0
`
`5.2
`
`5.1
`
`5.0
`
`4.8
`
`5.0
`
`7.3
`
`2.6
`
`2.7
`
`2.5
`
`2.5
`
`2.6
`
`1.7
`
`9.6
`
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`U.S. Patent
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`Sheet 10 of 10
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`US 7,763,717 B1
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`snnrption would partially alleviate the environrnental stress
`on the current marine environment.
`
`1
`CONTINUOUS PROTEIN AND LIPID
`RECOVERY FROM FOOD ANIMAL
`PROCESSING BYPRODUCTS
`
`RELA'l‘|?.D APPl..lCATlONS
`
`This application claims the benefit of U.S. Provisional
`Application Scr. No. 60r‘659,685 entitled “Continuous Pro-
`tein and Lipid Recovery From Food Animal Processing
`Byproducts" filed on 8 Mar. 2005, the contents of which are
`incorporated herein by reference in its entirety.
`
`BACKGROUND OF THE INVENTION
`
`Filleting fish requires removal of byproducts such as
`bones. skin. fin. scales, viscera and head. Most processors
`fillet fish by mechanical means. Mechanical filleting of one
`hundred pounds of trout
`(oncorlzyrrchus rnykisr} yields
`approximately forty pounds of fillets and sixty pounds of
`byproducts. The byproducts contain approximately twenty
`pounds of meat, which is half the amount of the fillets, and
`five pounds of fish oils (lipids). The byproducts are primarily
`land-filled, or ground and discarded. ln descriptive tcmis, per
`two truckloads of trout fillets going to the market. one truck-
`load of trout meat and a quarter of a truckload of trout lipids.
`which are not recovered from the byproducts. are land-filled,
`or ground and discarded.
`Historically fish byproducts have not been fully utilized by
`t.he rendering industry due to the “fishy odor" caused by
`auto-—oxicla1ion of fish oil. The odor is transferred to the meat
`of animals fed excessive amount of fish in their diets, result-
`ing in lower meat quality, and thus. limited consumer accep-
`tance. Free radicals. normally generated during the auto-
`oxidation. further deteriorate other components of animal
`feeds such as proteins, vitamins and the like. Fish processors
`incur expenditures to remove processing byproducts from
`their facilities. These byproducts are also a significant envi-
`ronmental bio-burden.
`Mechanical filleting of other fish species yields even less
`fillets and more byproducts. Mechanical filleting of one hun-
`dred pounds of tilapia {0reochmrm‘s
`tritlotictrs) yields
`approximately thirty pounds of fillets and seventy pounds of
`byproducts, resulting in even higher amounts of fish meat and
`oil being disposed of on per fish basis. Species such as Atlan-
`tic menhaden [Brevoortfa ryrarmus) are regarded as low-
`value species due to high amounts and distribution of bone,
`and high concentration of lipids. Fish species that have char-
`acteristics similar to menhaden are underutilized, or not uti-
`lized. for human consumption due to the unavailability of a
`proper meat recovery technology that can efiiciently elimi-
`nate the bones and lipids from the fillets.
`Antarctic krill (Euplzausia ssrpcrha) are small, shrimp-like
`crustaceans in the seas with the largest biomass of any multi-
`cellular animal species on earth. Estimates stale that one
`hundred filly million metric tons of krill could be an annual
`sustainable harvest compared with one hundred million met-
`ric tons ofthe total global seafood human consumption. Small
`krill size and endogenous proteases are processing chal-
`lenges, however, which have resulted in the failure of com-
`mercial krill fisheries for human consumption. According to
`the Food and Agricultural Organization (FAO), Atlantic and
`Pacific fish stocks have been exceeding the maximum sus-
`tainable levels since 1980 and 1999, respectively. Current
`commercial catch results in over-fishing and should be low-
`ered to approximately eighty million metric tons. Utilization
`of fish meat and lipids recovered from fish filleting byprod-
`ucts, krill. and species such as menhaden for human con-
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`The growth of the aquaculture industry encourages the
`development of technologies that recover proteins and lipids
`from filleting byproducts, and increases the total return.
`Existing surimi technology could he a good alternative for
`recovery of functional proteins; however,
`the traditional
`surimi processing cannot recover proteins from the byprod-
`ucts and uses excessively large volurues of water. Surimi is
`de-boned and skinned fish; the fillets are minced, washed and
`finally strained to form a concentrated fish paste.
`Surimi is an ancient process to make a protein food pre-
`dominantly derived from fish. Water is used in the process for
`making surinii. and can be used in a ration from about two
`parts water to one part fish up to about five parts water per one
`pan fish; typically. three parts water is used per one part fish.
`Two to five washes are used. Twenty to thirty percent of the
`fish muscle proteins are soluhilized when the ground muscle
`is washed with water. These soluble proteins, known as sar-
`coplasmic proteins, are generally not recovered from the
`wash water of the surirni process. These solubilized proteins
`are a good source of protein for animal or human feedstock.
`Only minced proteins, typically fish muscle proteins, are used
`in the surinii . The resultant washed minced protein product, in
`solid form, is then processed further to make protein gels.
`Kamboko is a popular fish sausage, produced by the surimi
`process, in which the washed minced fish is heated until it
`gels. High quality surimi is generally only produced from
`lean white fish. About fifty to sixty percent of the total protein
`of the muscle tissue is lost with dark-fleshed fish sources.
`Newer methods have been derived in an elTort to extract
`edible protein from muscle sources. U.S. Pat. Nos. 6.005,0'r'3
`(‘ 073) and 6,288.216 (’2l6) issued to Hultin et al ., on Feb. 12,
`1997 and on Sep. 1 l, 2001 respectively, disclose a process for
`isolating a protein composition front a muscle source and
`protein composition by mixing a particulate form of the
`muscle with an acidic aqueous liquid having a pH below
`about pH 3.5 to produce a protein rich solution. A protein rich
`aqueous solution is separated from solids and lipids, includ-
`ing membrane lipids. The protein rich aqueous solution can
`be treated to effect protein precipitation, followed by protein
`recovery. Furthermore, the inventions, of the ‘O73 and ‘Z16
`patents, require frequent water replacement. The particulate
`form of muscle is pre-prepared from muscle that has already
`been separated from most bone and other byproducts.
`U.S. Pat. No. 6.451.975 (’975} also issued to I-Iultin et al.
`on Sep. 1?, 2002 discloses a protein composition and process
`for isolating a protein composition ii-om a muscle source by
`mixing a particulate form of the tissue with an acidic aqueous
`liquid having a pH below about pll 3.5 to produce a protein
`rich solution substantially free of myofibrils and sarcomere
`tissue structure. The protein rich aqueous solution can be
`treated to effect protein precipitation. followed by protein
`recovery. U .8. Pat. No. 6,136,959 (‘ 959) is sued to Hultin et al.
`on Oct. 24, 2000 describes an alkaline protein extraction
`process which isolates edible protein from animal muscle by
`solubilizing the protein in an alkaline aqueous solution. The
`resultant solution contains 1 5% or less animal muscle. Again
`the muscle is pre-prepared fiom muscle that has already been
`separated from most bone and other byproducts.
`U.S. Patent Application No. 2003;’ 124,239 applied for by
`Kelleher on Feb. 19, 2003 describes a water soluble peptide
`composition, also derived from animal muscle tissue pro-
`teins. An enzyme is utilized in the process to make the peptide
`composition, and the resultant peptide composition contains
`less than about one weight percent fats and oils based upon
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`US 7,763,717’ Bl
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`the weight ofthe peptide composition and less than about two
`weight percent ash based on the weight of the peptide cont-
`position.
`U.S. Patent Application No. 2004t'067.55l, PCT applied
`for by [-Iultin et al. on Sep. 5, 2001, describes a protein
`extraction process for isolating edible protein lrorn animal
`muscle by solubilizing the protein in an alkaline aqueous
`solution. Undesirable components such as bones. neutral lip-
`ids, membrane lipids, fatty pieces, skin, cartilage, and other
`insoluble material are removed and discarded.
`
`U.S. Patent Application No. 2005;‘233_.060 applied for by
`Kelleheron Sep. 5. 2003 discloses a functional animal muscle
`protein concentrate composition and process for making the
`protein concentrate composition. The concentrated aqueous
`acidic protein solution derived from animal muscle tissue is
`added to the meat or fish prior to cooking. Similarly. U 3. Pat.
`No. 6,855,364 issued to Kelleher et al. on 1-"eh. 15, 2005
`describes a process tor retaining moisture in cooked animal
`muscle which involves adding a dry protein mixture or an
`aqueous acidic protein solution derived from animal muscle
`tissue to meat, including fish. prior to cooking.
`All of these processes take advantage of low protein solu-
`bility at their isoelectric point. It is well known in the art to use
`low protein solubility at their isoelectric point to isolate pro-
`teins. Furthermore. these processes produce peptides. which
`are products of a hydrolytic breakdown of proteins.
`
`SUMMARY OF THE INVENTION
`
`The present invention relates to a novel process for recov-
`ering lipids and protein from food animal processing byprod-
`ucts, and is especially useful in the recovery of functional
`muscle proteins, lipids and processing byproducts from fish.
`A batch operation and a conti.nuous operation for protein and
`lipid recovery that allows an efficicnt recovery of functional
`muscle proteins and lipids from food animal processing
`byproducts are included in the present inventiolt.
`The batch operation,
`tuilike the continuous operation
`modes is defined as a cyclic operation that requires repetitive
`cycles of loading substrate such as fish or fish-processing
`byproducts. processing involving isoelectric soIubili7ation
`and precipitation of fish muscle proteins, and unloading the
`products which may consist of recovered fish muscle proteins
`and lipids. In contrast to batch operation. continuous opera-
`tion mode allows continuous feeding of substrate, continuous
`processing, and continuous harvest of the products. The
`present continuous operation mode is a useful operation type
`for Lhe protein and lipid recovery from fish or fish processing
`byproducts.
`Using trout as an example, the protein recovery yield of the
`present invention is approximately 90% on dry weigh basis.
`The recovery is based upon isoelectric solubilization and
`precipitation of trout muscle proteins. The recovered trout
`muscle proteins retain their functionality, gelation, which is
`critical
`in development of restructured value-added food
`products. The laboratory-developed gels mimic restructured
`value-added foods and allow scientific determination of tex-
`ture and color properties, which are two important quality
`attributes for these foods.
`
`An isoelectric solubilizationfprecipitation of fish muscle
`proteins is applied to isolate functional proteins. This tech-
`nique oflers several advantages including high yield, separa-
`tion of impurities (bones. skin and scales) and a continuous
`mode of operation. enabling water recycling without treat-
`ment. This procedure may bring significant benefits to both
`the fish industry and environmental protection.
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`The proteins are solubilized at either acidic (about 2.0 to
`about 3.5) orbasic (about 10.5 to about 13.0) pH. followed by
`removal of insoluble materials with subsequent protein pre-
`cipitation at their isoelectric point (pH 5.5). followed by their
`separation from water. The muscle proteins recovered from
`trout retain their functionality—ge]ation._ which is critical in
`development of restructured va1ue—adcled food products. The
`omega-3 fatty acids included in the lipids recovered from
`trout do not exhibit degradation due to the pl-I treatment
`during protein and lipid recovery.
`Five steps are used to recover muscle protein and lipids,
`according to the present invention: 1. homogenization that
`simplifies sample handling and increases surface area of pro-
`teins and lipids, and therefore facilitates interaction between
`_ proteins and lipids with water: 2. first pH shift {front about pH
`2.0 to about pH 3.5. or from about pH 10.5 to about pI-] 13.0)
`that results in protein solubilization due to increased electro-
`static interaction between proteins and water, and facilitates
`separation oflipids from waterdue to increased polarity ofthc
`solution; 3. separation by first centrifugation. at about
`l0,000><g for about ]0n1inutcs at about 4° C. for batch pro-
`duction in the laboratory and at about 3,000 to about 3,5(}0><g
`for about 2 .0 to about 2.5 minutes at about 2° C . to about 9° C.
`in a continuous centrifuge (decanter) for continuous produc-
`tion, that results in bottom fraction of the fat-free impurities
`such as, bones. skin, fins. and insolubles. middle fraction of
`muscle proteins solubilized in water. and top fraction of fish
`lipids rich in omega-3 fatty acids as confirmed by experimen-
`tation: 4. the middle fraction is recovered and subjected to the
`second pl] shift at the proteins’ isoelectric point that results in
`isoelectric precipitation of muscle proteins due to decreased
`electrostatic interaction between proteins and water and
`increased hydrophobic interaction between proteins; and 5.
`separation by second centrifugation, at about 10.000xg for
`about 10 minutes at about 4° C. for batch production in the
`laboratory and at about 3.00010 about 3,S00xg for about 2.0
`to about 2.5 minutes at about 2° C . to about 9° C.
`in a
`continuous centrifuge (decanter) for continuous production,
`that results in separation of precipitated ftuictioual muscle
`proteins from water. The water separated in this step is pro-
`tein-free and clear and therefore can be recycled in the con-
`tinuous process. The continuous system for protein and lipid
`recovery is based on byproduct homogenizer (step one), two
`bio-reactors (steps two and four) and two continuous centri-
`fitges (steps five and six).
`A protein and lipid recovery in a continuous tuode accord-
`ing to the present invention uses the same ha sic five steps: first
`step, Homogcnization; second step. First pH Shift: third step,
`First Centrifugation; fourth step. Second pH Shift; and fifth
`step, Second Centrifugation. The homogenization of the fish
`processing byproducts with water is accomplished by using a
`continuous meat homogenizer such as for example Stephan
`Microcut MC}-l-10. The ltttittogenized slurry is continuously
`pumped using a peristaltic ptunp to the first bioreactor such as
`for example New Brunswick Scientific BioFlo 110 for the
`first pH adjustment (about pH 2.0 to about pl] 3 .5 or about pH
`10.5 to about pH 13.0). The soluble proteins and lipids are
`separated from the insolubles using a continuous separator
`such as for example-Alfa Laval MRNX 438 DD decanter. The
`separated soluble proteins and lipids are pumped to the sec-
`ond bioreactor for the second pH adjustment (pll 5.5) to
`precipitate the muscle proteins at their isoelectric point. The
`precipitated proteins are separated from the water and lipids
`by the second decanter. The separated proteins can be mixed
`with cryoproteclants and antioxidants if required and frozen
`for storage or used immediately to develop value-added food
`products. The water is re-used in the first step (i.e._. homog-
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`enization). This system can work continuously at a flow rate
`of 120 Lfhr. The How rate cart be modified by scaling-up the
`equipment.
`The protein and lipid recovery technology, according to the
`present invention, has several apparent and multi-"fold aspects
`including environmental, food production and economical.
`The present invention is more environmentally responsible.
`Less waste results from the recycling of water in the continu-
`ous mode. Less waste, where fish is the source of the protein,
`results from the fact that the present invention extracts more
`protein from the rendered fish than theprior art which extracts
`the protein solely from large muscles. Less environmental
`stress associated with the disposal of the processing byprod-
`ucts and over-fishing of depleted marine stocks results where
`the present
`invention is used with fish. Furthermore,
`the
`prices of fish products are lower, and a bigger variety of
`nutritious aquatic food products are possible when the present
`invention is used with fish as the protein source.
`The muscle proteins and lipids can be efficiently recovered
`from otherwise useless food animal processing byproducts.
`The byproducts often pose an enviromnental hazard when
`disposed of conventionally. "the recovered proteins, lipids.
`and fat-free impurities can be made into human food prod-
`ucts, pet food, animal feed. and plant fertilizer.
`Animal species not utilized for human consumption at
`commercial scale yet abundantly available such as Antarctic
`krill (Eaphrmsia super.-Ba) and Atlantic menhaden (Ber'voor-
`no ryrarrrrrrs} due to the lack of efficient protein and lipid
`recovery technologies can potentially be used for develop-
`ment of human food products.
`The present invention may be used in fish, poultry, and red
`meat processors, human food product development, nutra-
`ceutical industry. pharmaceutical industry. cosmetics indus-
`try, dietary supplements industry, animal feed industry, pet
`food industry and plant fertilizer industry.
`An aspect ofthe present invention is the use of food indus-
`try proccs sing byproducts as starting material and not animal
`muscle. In addition to processing byproducts, the present
`invention may use animal muscle, krill, fish, poultry. shrimp,
`and pork as starting materials.
`Another aspect of the present invention is that, although
`byproducts are used as starting material, the resultant proteins
`are ash-free. In other words, the resultant proteins lack bone,
`skin, and the like. These proteins are in a fomi of a viscoelas-
`tic paste, and exhibit bothviscous and elastic properties when
`deformed. The proteins may be processed further into gels.
`Yet another aspect of the present invention is to recover
`additional products, namely, oil (lipids) and the fat-free impu-
`rities. Similarly to the proteins recovered by the present
`invention. the lipids do not undergo degradation. The fat-free
`impurities are a fat-fi-ee product which is rich in growth-
`promoting minerals as well as non—muscle proteins. The fat-
`free impurities could optionally be isolated for use in animal
`and pet feeds.
`Another aspect ofthe present invention is that the proteins
`are separated from water. and the water is recycled in the
`continuous mode of the present invention. The processing
`water is a significant issue in the food processing industries.
`Pollution and wastewater treatment are unfortunate features
`of the food processing industries. Minimizing the amount of
`water released into the environment or in need of treatment
`reduces both cost and environmental impact of the industrial
`process.
`A further aspect of the present invention is the continuous
`mode aspect, which facilitates a cyclic flow through with
`substrate fed in and products harvested in a continuous man-
`ner unlike a traditional linear batch operation. The resultant
`
`system offers protein recovery at -90% or greater. Further-
`more, the continuous system also offers faster proc cssing, and
`therefore, less protein and lipid degradation occurs than when
`using conventional technology. The present invention allows
`processing times of thirty (30) minutes or less. A short pro-
`cessing time, due to increased processing efliciency, limits
`degradation of the products. The quality, of the resultant
`recovered proteins and lipids, is increased thereby.
`An aspect of the present invention is the separation follow-
`ing protein solubilization produces three distinct phases. The
`products include protein solution,
`lipids, and the fat—free
`impurities. Peptides are not a significant product of the
`present invention.
`Anotlter aspect of the invention is that we flocculate pro-
`teins following their precipitation. which allows the use of
`continuous decanters with lower g force instead of high speed
`centrifuges. Lower g force prevents excessive foaming that
`makes subsequent protein separation prohibitive. Further-
`more. to facilitate lipid separation from the protein solution
`following protein solubilization, emulsion breakers may be
`used to allow lower g forces, and prevent excessive foaming.
`The bio-reactors have specially designed mixing baffles
`and vessel shape to prevent pH gradient and prevent excessive
`foaming. The bio-reactor vessels and the mixing baffles are
`manufactured by Sartorius BB1 Systems of Bethlehem, Pa.
`The pH at which the proteins are solubilized in the present
`invention makes the proteins
`soluble and significantly
`reduces solution viscosity. The reduced viscosity facilitates
`subsequent continuous decanting.
`These and other aspects of the present invention will
`become readily apparent upon further review ofthe following
`drawings and specification.
`
`BRIEF Dl3SCRlP"I‘lOl‘l OF Tl Ill‘. DRAWINGS
`
`The novel features of the described embodiments are spe-
`cifically set forth in the appended claims; however, embodi-
`ments relating to the structure and process of making the
`present invention, may best be understood with reference to
`the following description and accompanying drawings.
`FIG. 1 is a flow chart showing a process ofprotein and lipid
`recovery from animal byproduct material according to the
`present invention.
`FIG. 2 is a How chart showing an example of a batch
`operation according to the present invention
`FIG. 3 is a flow chart showing an example of a continuous
`operation according to the present invention.
`FIG. 4 is a diagram depicting an example ofa system set up
`to carry out a continuous operation according to the present
`invention.
`
`FIG. Sis a flow chart showing as embodiment of the pro-
`cess of protein and lipid recovery from animal byproduct
`material, according to the present invention.
`FIG. 6 is a graph plotting time versus optical density at 595
`run after flocculation of fish muscle proteins subjected to high
`molecular weight anionic flocculent and isolated by isoelec-
`tric solubilizationfprecipitation
`FIG. 7 is a graph plotting time versus optical density at 595
`run after flocculation offish muscle proteins subjected to high
`molecular weight anionic flooculent and isolated by isoelec-
`tric soiubilizationfprecipitation
`FIG. 8 is a graph plotting time versus optical density at 595
`nm afier proteins were subjected to low molecular weight
`anionic fiocculent.
`
`FIG. 9 is a graph demonstrating protein recovery yields.
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`FIG. 10 is a graph demonstrating protein solubility indi-
`cating that proteins solubilizc and precipitate in water as a
`function of pH.
`