The effect of acid and alkali unfolding and subsequent refolding on the pro-oxidative activity of trout hemoglobin.
The pro-oxidative activity of trout hemoglobin was significantly increased at low pH (2.5-3.5) in a washed fish muscle (WFM) system. It was found that the more unfolded the hemoglobin was the more exposed its heme group was, which increased its pro-oxidative activity. The amount of oxidation products produced (TBARS) were, however, lower at low pH vs neutral pH. At pH 10.5-11, the pro-oxidative activity of hemoglobin was greatly suppressed. The conformation of hemoglobin was significantly more stable at high pH as compared to pH 7 as judged by its visible absorption spectrum. Hemoglobin readjusted from low pH to pH 7 had a higher pro-oxidative activity (i.e., more rapid oxidation) in WFM than native hemoglobin at pH 7, even though TBARS values were lower than in the untreated sample at pH 7. The results suggest that the WFM becomes slightly more susceptible to oxidation after low pH treatment but also produces less TBARS. The increased pro-oxidative activity after pH readjustment correlated well with an incomplete recovery in the native structure on pH readjustment. A longer unfolding time and a lower pH led to a less refolded hemoglobin with increased pro-oxidative activity. Hemoglobin was less pro-oxidative at low pH in the presence of 500 mM NaCl. The presence of salt did, however, increase the pro-oxidative properties of hemoglobin after readjustment to pH 7. The treatment of washed fish muscle at alkaline pH followed by adjustment to pH 7 led to a slight delay in hemoglobin-mediated lipid oxidation in WFM as compared to native hemoglobin at pH 7. The results suggest that WFM becomes less susceptible toward oxidation after pH readjustment from alkaline pH. These results clearly show that for muscle protein extraction/isolation processes requiring highly alkaline or acidic conditions, alkaline conditions are preferred if the lipid oxidation originating from hemoglobin is to be minimized.
Laboratory of Aquatic Food Biomolecular Research, Aquatic Food Products Program, Department of Food Science and Human Nutrition, University of Florida, Gainesville, Florida 32611, USA. hgkristinsson@mail.ifas.ufl.edu
Hordur G
HG
Kristinsson
Herbert O
HO
Hultin
D000818:Animals; D006454:Hemoglobins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D009132:Muscles / Q000737:chemistry; D016877:Oxidants / Q000032:analysis*; D010084:Oxidation-Reduction; D017510:Protein Folding; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D014337:Trout / Q000097:blood*
D006454:Hemoglobins; D016877:Oxidants; D017392:Thiobarbituric Acid Reactive Substances