Enzymes lactoperoxidase

Control experiments should be included in order to determine whether the cells under investigation contain endogenous systems capable of catalyzing iodination or iodide oxidation. These controls should include a sample in which the enzyme lactoperoxidase is omitted and one to which peroxide is not added. 

Nitric oxide and nitrite react with other peroxidase enzymes such as horseradish peroxidase (HRP) (138a,139), lactoperoxidase (138a) and eosinophil peroxidase (140) similarly. The rate constants for reaction of NO with compounds I and II in HRP were found to be 7.0 x 105M 1s 1 and 1.3 x 106M 1s 1, respectively (139). Catalytic consumption of NO as measured by an NO sensitive electrode in the presence of HRP compounds I and II is shown in Fig. 5 where accelerated consumption of NO is achieved even in deoxygenated solutions (140). 

Since hemoproteins such as lactoperoxidase and catalase are inhibited more rapidly than the sulfhydryl oxidation occurs, it is unlikely that the rapid activation of guanylate cyclase occurs by sulfhydryl oxidation [132]. Prolonged incubation of the papain or dehydrogenase enzymes with substrate and nitroprusside yielded a turbidity which indicated denaturation of the enzyme to an insoluble form, possibly by the formation of disulfide bridges via the dimerization of thiyl radicals [132]. 

With a few exceptions (e.g. lysozyme and lactoperoxidase), the indigenous milk enzymes do not have a beneficial effect on the nutritional or organoleptic attributes of milk, and hence their destruction by heat is one of the objectives of many dairy processes. 

The occurrence of a peroxidase, lactoperoxidase (LPO), in milk was recognized as early as 1881. It is one of the most heat-stable enzymes in milk its destruction was used as an index of flash pasteurization (now very rarely used) and is now used as an index of super-HTST pasteurization. 

Since the principal constituents of milk are proteins, lipids and lactose, proteinases, lipases and / -galactosidase (lactase) are the principal exogenous enzymes used in dairy technology. Apart from these, there are, at present, only minor applications for glucose oxidase, catalase, superoxide dismutase and lysozyme. Lactoperoxidase, xanthine oxidase and sulphydryl oxidase might also be included, although at present the indigenous form of these enzymes is exploited. 

Bjorck, L. (1993) Indigenous enzymes in milk Lactoperoxidase, in Advanced Dairy Chemistry, Vol. 1 Proteins, (ed. P.F. Fox), Elsevier Applied Science, London, pp. 332-9. 

The activity of selected enzymes is used as indices of thermal treatments, e.g. alkaline phosphatase (HTST pasteurization), y-glutamyl transpeptidase (index of heating in the range 72-80°C) or lactoperoxidase (80-90°C). 

Peroxidases. Another group of enzymes, which is involved in the oxidation of xenobiotics, is the peroxidase. There are a number of these enzymes in mammalian tissues prostaglandin synthase found in many tissues, but especially seminal vesicles and also the kidney, the lung, the intestine spleen, and blood vessels lactoperoxidase found in mammary glands myeloperoxidase found in neutrophils, macrophages, liver Kupffer cells, and bone marrow cells. 

The human body contains lactoperoxidase, a product of exocrine secretion into milk, saliva, tears, etc., and peroxidases with specialized functions in saliva, the thyroid, eosinophils,219 and neutrophils.220 The functions are largely protective but the enzymes also participate in biosynthesis. Mammalian peroxidases have heme covalently linked to the proteins, as indicated in Fig. 16-12 220 222a... 

The direct interactions between metals and ONOO- can catalyze modifications. For example, the metals in Cu,Zn SOD and FeEDTA (EDTA = ethyl-enediaminetetraacetic acid) enhance nitration reactions (229). Heme-containing enzymes such as myeloperoxidase (6 x 106A/-1 s-1) and lactoperoxidase (3.3 x 105M-1s-1) also react with ONOO- (230) such that compound II [FeIV(P+)0] is formed. In contrast, horseradish peroxidase (3.2 x 106M-1 s-1) is converted to compound I (FevO) by ONOO-. Floris et al. (230) proposed an interesting mechanism by which compound I is initially produced and then rapidly oxidizes NO-f to N02. In the presence of NO, a number of nitrosation reactions would subsequently be facilitated by subsequent formation of N2O3 (Eq. 32). 

Griffiths (2) studied the thermal stability of the naturally occurring enzymes in milk at temperatures between 65 and 80°C, in order to choose an enzymatic index of adequate pasteurization of milk. For the acid phosphatase, the D-values of 7.38 min at 75°C and 7.87 min at 80°C corresponded to a z-value of 6.6°C for the inactivation of lactoperoxidase, D-values of 0.80 min at 75°C and 0.075 min at 80°C corresponded to a z-value of 5.4°C for amylase (saccharifying activity), D-values of 0.85 min at 75°C and 0.45 min at 80°C corresponded to a z-value of 16.2°C. The naturally occurring enzymes in milk showed lower D-values than those found for GFPuv in the three buffers. However, a z-value of 16.64°C characterized for GFPuv in phosphate buffer solutions at pH 7.0 was similar to that obtained for amylase in milk. 

Fig. 2.7 Detail of the reconstructed phylogenetic tree showing the subfamily of vertebrate peroxidases including the mammalian enzymes myeloperoxidase (MPO), eosinophil peroxidase (EPO), lactoperoxidase (LPO), and thyroid peroxidase (TPO). Reproduced from [10] with the permission of John Wiley and Sons (License Nr. 2326000554179)...

Sulphydryl oxidase, an indigenous milk enzyme, has been proposed for the oxidation of thiols in UHT milk to reduce cooked flavor and also thereby to serve as an antioxidant, in conjunction with lactoperoxidase (to destroy the resultant H2O2), by obviating pro-oxidants resulting from autoxidation of thiols (Swaisgood and Abraham, 1980). 

The mechanism(s) by which Xanthine oxidoreductase exerts its prooxidant effect(s) is not fully understood. Hydrogen peroxide, resulting from oxidation of a suitable substrate by Xanthine oxidoreductase, could oxidize milk lipids. However, normal milk contains little or no substrate for the enzyme. A possible mechanism involving interaction between native and denatured Xanthine oxidoreductase in MFGM and lactoperoxidase or copper in milk serum has been proposed (Hill, 1979 Allen and Wreiden, 1982b). 

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