In this review, recent findings related to various factors influencing quality properties of fish meat and its products during frozen storage are introduced. Many studies have indicated that protein denaturation is the factor determining the quality of frozen fish meat. Ice crystal size does not necessarily determine the quality of frozen fish meat because the tissue of meat reabsorbs water during the thawing process, unless it has been previously damaged by protein denaturation. However, the effects of ice crystals on the quality of thawed fish meat differ based on the fish species, post-mortem stages, protein denaturation, and processing conditions of the fish meat. In the case of frozen-thawed lightly salted fish meat, salting conditions greatly affect the water holding capacity of muscle and the ice crystal size. Also, in the case of frozen kamaboko, which is denatured protein gel, as the thawed water is not absorbed enough by the protein gel, ice crystal size could be a determining factor of quality. The appropriate freezing and storage conditions required for maintaining quality must be based upon the characteristics of each seafood product.
A mild and efficient procedure for conjugating rabbit Fab' and horseradish peroxidase using a maleimide compound was developed. The enzyme was treated with N-hydroxysuccinimide ester of N-(4-carboxycyclohexylmethyl) maleimide to introduce maleimide groups. Then, the maleimide-enzyme was allowed to react with thiol groups of Fab', and the conjugate formed was separated from unreacted components by gel filtration with Ultrogel AcA 44. In the peak fraction of the separated conjugate, 98% of peroxidase was associated with Fab' and 90% of antibodies was associated with peroxidase. The recoveries in the conjugate of peroxidase and Fab' incubated for conjugation were 65-74%. The conjugate formed appeared to be largely monomeric. Both the enzyme activity and antigen-binding activity of Fab' were fairly well preserved in the conjugate. The cross-link formed was stable at 4 degrees C at least 6 months. Use of the conjugates obtained by this method gave greater sensitivity in sandwich enzyme immunoassay for human ferritin and human thyroid-stimulating hormone than conjugates prepared by the periodate method. The conjugation using N-hydroxysuccinimide ester of m-maleimidobenzoic acid provided a similar monomeric preparation but was less efficient.
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