Abstract
Laccases are blue multi-copper oxidases with an extensive number of actual and potential industrial applications. It is known that laccases from different fungal strains may vary in properties; however, the reason of this remains unclear. In the current study we have isolated and characterized seven laccases from different strains of Steccherinum ochraceum obtained from regions of central Russia. Although all seven laccases had the same primary sequences, there was a little variation in their molecular weights and thermostabilities. Moreover, statistically significant differences in laccases’ catalytic parameters of oxidation of phenolic substrates and ABTS were observed. After the deglycosylation of four selected laccases by Endo H and PNGase F, their affinities to pyrocatechol and ABTS became the same, suggesting a substantial role of N-linked glycosylation in moderation of enzymatic properties of laccases.
Highlights
Laccases (p-diphenol:dioxygen oxidoreductase, EC 1.10.3.2) are blue multi-copper oxidases that catalyze oxidation of a wide range of phenolic and non-phenolic aromatic compounds, coupled with a concomitant reduction of molecular oxygen to water [1,2]
In order to obtain sufficient for further comparative study amounts of laccases, seven strains of S. ochraceum were cultivated by a submerged method on a liquid glucose-peptone medium supplied with CuSO4 as an inducer
The cultural broth was collected at the 20–25th day of cultivation, when the laccase activity reached the maximum
Summary
Laccases (p-diphenol:dioxygen oxidoreductase, EC 1.10.3.2) are blue multi-copper oxidases that catalyze oxidation of a wide range of phenolic and non-phenolic aromatic compounds, coupled with a concomitant reduction of molecular oxygen to water [1,2]. In the last decade several reports demonstrated that different strains of the same species can produce laccases with different properties and complementary biochemical features [20,21]. All these reports did not directly address the cause(s) of observed variations. Steccherinum is a genus of basidiomycete fungi belonging to the Steccherinaceae family (order Polyporales) Some members of this family—Antrodiella faginea, Junghuhnia nitida, Steccherinum murashkinskyi, Steccherinum ochraceum and Steccherinum bourdotii–were previously reported as fungi with high laccase-producing capability [22,23,24]. In this study seven novel laccases from seven strains of Steccherinum ochraceum were purified and biochemically characterized; the nucleotide sequences of genes encoding these laccases were determined; and the influence of glycosylation on their catalytic properties was assessed
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