Bromoperoxidase isolation

Hara 1, T Sakurai (1998) Isolation and characterization of vanadium bromoperoxidase from a marine macroalga iicHowia stolonifera. J Inorg Chem 72 23-28. 

Vanadium bromoperoxidase (V-BrPO) has been isolated from many species of marine brown algae, including A. nodosum [1,26,27], Laminaria saccharina [28], Fucus distichus [29], and Macrocystis pyrifera [29] the red algae Ceramium rubrum [30] and C. pilulifera [31] and a terrestrial lichen, Xanthoria parie-tina [32],... 

Two vanadium bromoperoxidases that differ in carbohydrate content [26,33] have been isolated from A. nodosum. The most abundant bromoperoxidase, V-BrPO-I, was found in the thallus, and the other bromoperoxidase, V-BrPO-II, was reported to be present on the thallus surface [26], A previous report also concluded that V-BrPO is present in two different locations of A. nodosum, one in the cell walls of the transitional region between the cortex and medulla of the thallus and the other in the cell wall of the thallus surface [34], More recent experiments demonstrate that vanadium-dependent bromoperoxidase activity is present in both the cortical and surface protoplasts of M. pyrifera [35], L. saccharina, and L. digitata [36], The biosynthesis of V-BrPO in the protoplasts of L. saccharina has been shown using [35S]-methionine [36], The vanadium bromoperoxidases are all acidic proteins [26] with very similar amino acid compositions [37], V-BrPO (A. nodosum) has been crystallized, although refined structural data have not been reported yet [38], A different isolation procedure, based primarily on a two-phase extraction system, has been described [39,40], This procedure works well for certain types of algae (e.g., Laminaria) but not for the isolation of V-BrPO from A. nodosum, the principal source of V-BrPO for the mechanistic studies. 

Bromoperoxidase activity is expressed as micromoles of mcd brominated per minute per milligram of enzyme (U/mg). The early work on V-BrPO employed the oxidation of iodide by dihydrogen peroxide [1], forming triiodide (Ir), which was followed spectrophotometrically at 353 nm ( = 26,400 M 1cm 1). However, this reaction is less desirable for quantitation of haloperoxidase activity because of competing side reactions, such as the nonenzymatic oxidation of iodide by dihydrogen peroxide and reduction of triiodide by dihydrogen peroxide (discussed later). The specific activity of mcd bromination for V-BrPO isolated from A. nodosum is 170 U/mg (at pH 6.5, 2 mM H202, 0.1 M Br , 50 nM mcd,... 

There are several examples of haloperoxidases containing metals other than iron, and many of these are non-heme enzymes. For example, a series of bromoperoxidases have been isolated recently from seaweed17,18 and an actinomycete19 that require vanadium for halogenating activity. 

Bromoperoxidase has been isolated from Pseudomonas aureofaciens ATCC 15926, also displays peroxidase and catalase activities, and contains ferriprotoporphyrin IX (van Pee and Lingens 1985). Four different bromoperoxidases have been isolated from Streptomyces griseus (Zeiner et al. 1988). Only one of them, however, contains ferriprotoporphyrin IX and displays peroxidase and catalase activities. This illustrates that there are two different groups of enzymes, one of which lacks heme prosthetic groups. 

Vanadium ions take part in some an important processes occurring in biological systems [62]. One vanadium-containing enzyme, bromoperoxidase, has been isolated from several species of marine brown algae. This enzyme catalyzes the oxidation of the bromide anion by hydrogen peroxide, resulting in the bromi-nation of organic compounds ... 

The most intensively studied haloperoxidases are the chloroperoxidase from the mold Caldariomyces fumago [1322] and bromoperoxidases fi om algae [1764] and bacteria such as Pseudomonas aureofaciens [1765], Ps. pyrrocinia [1766], and Streptomyces sp. [1767]. The only iodoperoxidase of preparative use is isolated from horseradish root [1768]. A haloperoxidase isolated from milk has been reported to be useful for the formation of halohydrins [1769]. 

The new 6-bromo-icosa-3Z,5E,8Z,13E,15E-pentaene-11,19-diynoic add, isolated from an Indonesian Haliclom sp. suggests that a bromoperoxidase acts on the same C-6 position, even for nondemospongic adds (Aratake et al., 2009). 

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