Fish Protein Isolate Its Superior Gel Functionality

Transcription

1 Fish Protein Isolate Its Superior Gel Functionality June Rogers, Arkansas Jae W. Park Professor OSU Seafood Research & Education Center Astoria, OR 97103, USA

2 Content Introduction Processing Principles Processing Chemistry of Fish Protein Isolate (FPI) Extraction of FPI and its gelation Enzymes in FPI Lipids in FPI Color of FPI gels Superior Gelling Ability of FPI Conformation changes Role of sarcoplasmic proteins Disintegration of muscle tissue Commercial Applications of FPI Future of FPI Negative role of salt Conclusions

3 Introduction Surimi (Conventional): Refined and stabilized fish myofibrillar proteins. Refined means. Fish protein isolate (FPI) extracted by extreme ph: high acid/alkali ph the pi Adjust to the neutrality Stabilized with Cryoprotectants

4 Adapted from Park (2008)

5 DENATURATION MUST BE AVOIDED! DENATURATION IS INDUCED! 53 Adapted from Park (2008)

6 Publications on FPI

7 Publications on FPI

8 Extraction and Isolation of FPI

9 Homogenization Extraction Neutral Lipids Soluble Protein Centrifugation Sediment Bone, Membrane lipid, Connective tissue

10 Homogenization Extraction Neutral Lipids Soluble Protein Centrifugation Membrane lipids removed for Greater Stability

11 Homogenization Extraction Clean Effluent Precipitation Centrifugation FISH PROTEIN ISOLATE Freeze with Cryoprotectant

12 Significantly High Yield! 46 Ingadottir, 2004

13 Enzymes in FPI

14 ATPase Activity ATPase activity is not measurable for acid- or alkali-extracted FPI A large portion myosin light chains were lost (Choi and Park, 2002; Kristinsson and Hultin, 2003b). ATPase activity is not a prerequisite for the gelation of FPI. Properties (Choi and Park, 2002) Conventional surimi processing 1-washing 3-washing FPI by Acid Extraction Ca-ATPase activity (μ mole Pi/min/mg protein) Mg-ATPase activity (μ mole Pi/min/mg protein) 0.128± ±0.003 ND 0.218± ±0.022 ND

15 Proteolytic Protease Activity Cathepsins are very reactive enzymes causing gel softening in many types of fish species Cathepsin L-like enzymes were found at all phs (Kim et al., 2003). Cathepsin B-like enzymes were highly activated at acidic phs (Kim et al., 2003).. L-like B-like

16 Transglutaminase Activity TGase is unique to fish proteins and makes a covalent bond upon slow heating or setting. Kim and Park (2008) reported TGase activity was reduced with FPI extracted by acid or alkali (see the next slide).

17 Conventional Surimi Effect of Acid or Alkali Extraction on TGase (Set Gel) Alkali FPI Acid FPI Non-setting Setting (Kim and Park, 2008)

18 Superior Gelling Ability of FPI Why? 40

19 Superior Gelling Properties of FPI compared to Conventional Surimi Fracture Analysis Pacific Whiting Breaking force (g) Deformation (mm) Breaking force (g) Deformation (mm) 100 Con ph2 ph3 ph10.5 ph11 ph12 6 (Dr. Park s Lab, 2003)

20 Figure 2 Breaking force and deformation of rockfish muscle proteins prepared by various treatments. Adapted from Yongsawatdigul and Park (2004). M: mince; WM: washed mince (surimi), AC: acid-extracted; AK: alkaliextracted. Different letters (a-d or x-z) denote a statistical difference (P<0.05).

21 Conformational Changes?

22 Kristinsson and Hultin 2003b: The isolate manufacturing process induces a Conformational Change in the HMM region of myosin that allows for Better Charge Distribution on the surface of the protein, leading to a Molten Globule State.

23 Figure 4 - Thermal aggregation of native and refolded myosin as assessed by turbidity development at 350 nm. Protein concentration was ~0.45 mg/ml and samples were heated in a sealed cuvette at 1.5ºC/min. Adapted from Kristinsson and Hultin (2003a).

24 Figure 6 Viscoelastic behavior of cod myosin on heating and cooling. Protein concentration was 35 mg/ml. Samples were heating at 1.5ºC/min and cooled subsequently when the temperature reached 80ºC. Adapted from Kristinsson and Hultin (2003b).

25 Log G' (Pa) Storage Modulus WM M AK AC Cross-linking of myosin Egelandsdal et al. (1986) Denaturation of LMM Temperature (C) 33 Figure 5 Dynamic rheology thermograms of rockfish mince prepared by various treatments: WM, washed mince; M, mince; AK, alkali-extracted FPI AC, acid-extracted FPI. Adapted from Yongsawatdigul and Park (2004)

26 Yongsawatdigul and Park (2004) Alkali-extracted FPI exhibited a significant role of disulfide linkages through possibly enhanced interactions: 1)between myofibrillar and sarcoplasmic proteins 2)between MHC and actin

27 Figure 3 SDS-PAGE patterns of rockfish muscle proteins prepared by various treatments solubilized in buffer with and without β mercaptoethanol (βme). Adapted from Yongsawatdigul and Park (2004). S: standard molecular weight; M: mince; WM: washed mince (surimi), AC: acid-extracted; AK: alkali-extracted.

28 8 7 6 Paste Gel b 5 b b 4 b 3 b a 2 a a 1 0 M WM AC AK Changes of Sulfhydryl Group Samples by transformation of paste to gel Total SH(mol/10 g protein 30 M: mince; WM: washed mince (surimi), AC: acidextracted; AK: alkali-extracted. Yongsawatdigul and Park (2004).

29 Role of Sarcoplasmic Proteins

30 Surimi vs. FPI Surimi = Refined Fish Myofibrillar Proteins FPI = Homogenized Myofibrillar Proteins + Sarcoplamic Proteins

31 Higher myofibrillar protein content = Lower sarcoplasmic protein content = Better Gel Strength However, Several reports exist showing that sarcoplasmic proteins contribute positively to gel formation of myofibrillar proteins: Morioka and Shimizu, 1990 Morioka et al., 1992) Morioka and Shimizu, 1993 Nomura et al., 1995 Ko and Hwang, 1995 Yongsawatdigul and Park, 2004 Kim et al., 2005 Park and Park, 2007

32 Effects of SP addition to Pollock surimi on texture properties Breaking force (g) Deformation (mm) Breaking force (g) Deformation (mm) 0 Control Sucrose (2%) SP (2%) 0 Kim et al. (2005)

33 Higher myofibrillar protein content = Lower sarcoplasmic protein content = Better Gel Strength THEN, if not the above, how do we explain better gelation of surimi than for mince, and better gelation of isolates than for surimi?

34 Disintegration of Muscle Tissue 24

35 Better gelation of surimi than for mince, and Better gelation of isolates than for surimi #1 Sato and Tsuchiya (1992) observed that stronger, more deformable surimi gels correlated with a more homogeneous dispersion of proteins, when viewed by TEM

36 Better gelation of surimi than for mince, and Better gelation of isolates than for surimi #2 Wright and Lanier (2008) observed stronger, more deformable gels made from alkaliextracted FPI viewed by TEM, which is more homogeneously dispersed proteins. Maximum disruption/dispersion occurred at ph 11

37 Lipid in FPI

38 Fate of Lipids in Surimi and FPI Wet surimi bland in odor Dried surimi fishy odor indicate Conventional Surimi Process does not remove membrane lipids. Hultin et al. (2005) suggested: Divalent cations (Ca ++ or Mg ++ ) along with organic acid (i.e., citric acid) can easily separate membrane lipids through homogenization and high speed centrifugation.

39 FPI Desired Stability Reduced Oxidative Rancidity

40 Table 1--- Protein recoveries and lipid reductions using the conventional surimi process, acid-aided process and alkali-aided process to recover proteins from Atlantic croaker Conventional Acid-aided Alkali-aided Protein recovery 57.7%a 78.7%c 65.0%b Lipid reduction 16.7%a 38.1%b 68.4%c Means in each row having different superscript letters are significantly different (P < 0.05). Kristinsson and Liang (2006): Vol. 71, Nr. 5, 2006 JOURNAL OF FOOD SCIENCE

41 More lipid reduction Chen and Jaczynski (2007)

42 Colors of FPI Gels

43 Catfish Kristinsson et al (2005) Rockfish Yongsawatdigul and Park (2005)

44 Commercial applications of FPI 14

45 The Succulence System New technology for leveraging your protein resource instead of phosphates, add meat... Increase raw and cooked yields dramatically with cleaner label Restore succulence to frozen meats Reduce or eliminate phosphates Pre-cook yields equal to or better than phosphates Login Cook yields superior to phosphates Recover meat from trimmings as a soluble protein marinade Establish a NEW control of quality/consistency in intact meat cuts

46 Nutrilean The Fat Blocker

47 OIL UPTAKE WHEN FRIED PRAWN SQUID CHICKEN FRIED KAMABOKO BALL 資料 : 調理のためのベーシックデータ ( 女子栄養大学出版部 ) Souce:Basic data for cooking(joshi-eiyou-daigaku syuppanbu) Hachisu (Surimi Industry Forum - April 20, 2009)

48 10 Future of FPI

49 RESEARCH GROUPS have agreed that Alkaline extraction followed by pi Isolation delivers Superior Gelling Functionality.

50 Can FPI replace Conventional Surimi? 8

51 Conventional Surimi Effect of salt addition on gel texture Alkali FPI Acid FPI Non-setting Setting Kim and Park, 2008

52 Should FPI be stored frozen with cryoprotectants? At what ph? 200 Breaking force Deformation 15 Breaking force (g) Deformation (mm) 0 CS CS-F 5C 5C-F 5NC-F 7C 7C-F 7NC-F 0 (Thawornchinsombut and Park 2004)

53 Are chemicals (NaOH and HCl) used simply as a processing aid? 2 N NaOH ph 6.5 ph 11 ph N HCl ph N NaOH FPI 5 (Fish Protein Isolate)

54 NaOH + HCl = NaCl + H 2 O ph 6.0~6.5 ph 11.0 ph 5.5 ph CFR (a)(3)(ii)(b) - Substances that are added to a food during processing, are converted into constituents normally present in the food, and do not significantly increase the amount of the constituents naturally found in the food. Therefore, NaOH and HCl are used as a processing aid and no labeling is required.

55 FPI can be More Attractive for Nutritional Enhancement! Good news to special fish (i.e., catfish, tilapia), poultry, and red meats Lower Omega-6/Omega-3 ratio effectively by injecting omega-3 fish oil - FPI emulsion

56 Summary Proteins were chemically unfolded (denatured?) when extracted in acid or alkali. Upon the ph adjustment to 7, myosin partially refolded back. More disruption and homogenous dispersion improved gelation. Alkaline extraction gives better texture, reduced lipid content. No significant effect of frozen storage ph, but cryoprotectants must be added. Use FPI independently with conventional surimi due to their different behavior against salt FPI can be more attractive for nutritional enhancement.

57 Acknowledgement Former students and visiting Scientists: Dr. J. Yongsawatdigul Young Kim Dr. Yeung J Choi Dr. Jacek Jaczynski Dr. Supawan Thawornchinsombut Dr. Joodong Park

58 Thank You.. Astoria, OR