CHARACTERIZATION OF PROTEINASE RECOVERED FROM PACIFIC WHITING SURIMI WASH WATER

Pacific whiting surimi wash water (SWW) proteinase was recovered by ohmic heating, ultrafiltration, and freeze-drying. By these processes, 5.9-fold purification was achieved. The most efficient step was ohmic heating, which concentrated the proteinase by 4.8 fold. Specific activity of the recovered SWW proteinase on casein and Z-Phe-Arg-NMec was 28.2 and 0.17 U/mg protein, respectively. The SWW proteinase showed good hydrolytic activity towards casein, acid-denatured hemoglobin and myofibrils. Acidification increased specific activity on all substrates tested but reduced thermal stability. β-Mercaptoethanol, dithiothreitol and urea enhanced activity against Z-Phe-Arg-NMec. Proteinase activity on Z-Phe-Arg-NMec showed an optimum pH of 4.0. The recovered proteinase showed 18.5% residual activity after 7 week storage at 4C.     

Physicochemical Changes in Pacific Whiting Muscle Proteins during Iced Storage

No changes in actomyosin Ca²⁺-, Mg²⁺-, or Mg²⁺-Ca²⁺-ATPase activities were observed during iced storage of Pacific whiting fillets, but Mg²⁺-EGTA-ATPase increased with a loss of Ca²⁺-sensitivity. Surface hydrophobicity of actomyosin increased substantially within 2 days, but not total sulfhydryl (SH) content. During longer storage, the SH content decreased gradually, but surface hydrophobicity remained constant. Autolytic degradation products increased in fish muscle with storage time. Myosin heavy chain (MHC) was degraded by 45% within 8 days, but no noticeable difference was observed in actin. Results indicated that autolysis may be the main cause of physicochemical changes in Pacific whiting muscle proteins during iced storage.     

INHIBITION OF GEL WEAKENING OF THREADFIN BREAM SURIMI USING THAI LEGUME SEED PROTEINASE INHIBITORS

Partially purified proteinase inhibitors from cowpea (Vigna unguiculata (L) Wasp), pigeon pea (Cajanus cajan (L.) Millsp.) and bdmbara groundnuts (Voandzeia subterranea (L.) Thou) effectively inhibited sarcoplasmic modori‐inducing proteinase extracted from threadfin bream muscle in a concentration dependent manner. Incorporation of these proteinase inhibitors into threadfin bream surimi partially inhibited autolytic degradation and increased the gel force and deformation. Combination of setting and incorporating proteinase inhibitors from cowpea and bambara groundnut var. HY at the level of 30 Kcunits/g resulted in an increase in gel force and deformation by 60% and 26%, respectively. However, the lightness and whiteness of surimi gels decreased slightly when the proteinase inhibitor was added at a level of 30 kunits/g.     

ANTIOXIDATIVE ACTIVITY OF PROTEIN HYDROLYSATE FROM ROUND SCAD MUSCLE USING ALCALASE AND FLAVOURZYME

Antioxidative activity of hydrolysates from round scad (Decapterus maruadsi) muscle with degrees of hydrolysis (DH ) of 20, 40 and 60%, prepared using Alcalase (HA) or Flavourzyme (HF), was determined. At the same DH, HF exhibited a higher 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging activity and reducing power, but a lower Fe²⁺ chelating ability than HA. HF from isopropanol‐defatted muscle with 60% DH was extracted using different solvents, and hexane (E1), dichloromethane (E2), ethyl acetate (E3) and residual (R) fractions were obtained. Among all fractions, E2 and E3 exhibited the highest DPPH radical scavenging activity and reducing power. HF with 60% DH and E2 at 1,000 ppm exhibited antioxidant activity in linoleic oxidation and lecithin liposome systems, and the results were comparable to butylated hydroxytoluene (BHT) at 100 ppm. Therefore, type of proteinase, DH and defatting process prior to hydrolysis exerted an influence on the antioxidative activity of hydrolysates.     

PHYSICOCHEMICAL AND BIOCHEMICAL CHANGES IN WHOLE LIZARDFISH (SAURIDA MICROPECTORALIS) MUSCLES AND FILLETS DURING FROZEN STORAGE

The physicochemical and biochemical changes in whole lizardfish (Saurida micropectoralis) muscles and its fillets kept in air and under vacuum during frozen storage at ?20C for 24 weeks were investigated. The formaldehyde (FA) and dimethylamine contents increased with a concomitant decrease in trimethylamine‐ N‐oxide (TMAO) content as the storage time increased (P < 0.05). The Ca²⁺–adenosine 5′‐triphosphatase activity continuously decreased with a coincidental decrease in salt‐soluble fraction. The disulfide bonds were increasingly formed throughout the storage (P < 0.05). The surface hydrophobicity increased and reached the maximum at week 12 with a subsequent decrease up to the end of storage. In general, the higher changes were observed in samples kept under vacuum than those kept in air. With the same atmosphere used, the whole fish showed slightly higher changes than the fillets. A marked increase in TMAO demethylase (TMAOase) activities was observed up to 12 weeks, followed by the continuous decrease up to 24 weeks of storage. The produced FA might play an important role in inducing protein denaturation and/or aggregation in lizardfish. The TMAOase activity as well as the FA formation could be reduced to some extent with the removal of internal organs and storage in the presence of oxygen. However, a detrimental effect of oxygen, especially on the promotion of lipid oxidation, would be an obstacle.     

EXTRACTION OF CAROTENOPROTEIN FROM BLACK TIGER SHRIMP SHELLS WITH THE AID OF BLUEFISH TRYPSIN

The effect of bluefish (Pomatomus saltatrix) trypsin on the recovery and characteristics of carotenoprotein from black tiger shrimp (Penaeus monodon) shells was investigated. Trypsin concentration and reaction time both affected the hydrolysis and the recovery of carotenoproteins ( P <  0.05). The recovery of carotenoproteins from shrimp shells was maximized by the hydrolysis of shrimp shells using 1.2 trypsin units/g shrimp shells for 1 h at 25C. Freeze-dried carotenoprotein recovered contained 70.20% protein, 19.76% lipid, 6.57% ash, 1.50% chitin, and 87.91 µg total astaxanthin/g sample, indicating a substantial reduction in the levels of antinutrients associated with shrimp waste, while enriching the product in carotenoid pigments and valuable essential nutrients (proteins and lipids). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the recovered carotenoprotein revealed that protein with molecular weight of 45 kDa was the major constituent. When hydrolytic activities of bluefish and bovine trypsins toward carotenoproteins in black tiger shrimp shells were compared, the recovery efficacy of protein and pigment by bluefish trypsin was similar to that achieved by trypsin from bovine pancreas. Therefore, bluefish trypsin could be used as an alternative cheap proteinase for extraction of carotenoproteins from black tiger shrimp shells.

PRACTICAL APPLICATIONS

Carotenoproteins from black tiger shells, the byproduct of shrimp processing, can be recovered with the aid of fish trypsin. This product can be used for both food and feed applications. Additionally, the fish trypsin can be used instead of bovine trypsin. As a whole, the utilization of fish and shellfish processing wastes can be maximized.     

EFFECT OF CHICKEN PLASMA PROTEIN AND SOME PROTEIN ADDITIVES ON PROTEOLYSIS AND GEL-FORMING ABILITY OF SARDINE (SARDINELLA GIBBOSA) SURIMI

The effect of chicken plasma protein (CPP) and various protein additives on autolysis and gel‐forming ability of sardine (Sardinella gibbosa) surimi was investigated. CPP and other protein additives showed inhibitory activity toward autolysis of sardine surimi incubated at 70C in a concentration‐dependent manner. Porcine plasma protein (PPP) and egg white (EW) were more effective in proteolysis prevention than CPP and other protein additives. Breaking force and deformation of both modori and kamaboko gels increased when CPP and other protein additives were added at levels up to 2% (P < 0.05). Nevertheless, PPP and EW showed a greater gel‐strengthening effect than CPP and other protein additives (P < 0.05). Addition of CPP and other plasma proteins resulted in decreased whiteness, especially with increasing amount (P < 0.05). However, no change in whiteness was observed with gels containing EW and soy protein isolate (SPI) (P > 0.05). Proteolysis of sardine surimi can be retarded by the addition of CPP and protein additives, leading to increased gel‐forming ability.