Characterization and role of tyrosinases in the lichenLobaria pulmonaria(L.) Hoffm.

2016 ◽  
Vol 48 (4) ◽  
pp. 311-322 ◽  
Author(s):  
Lusanda P. MATEE ◽  
Richard P. BECKETT ◽  
Knut A. SOLHAUG ◽  
Farida V. MINIBAYEVA
Keyword(s):  
Enzyme Activity ◽  
Tyrosinase Activity ◽  
Melanin Synthesis ◽  
Direct Sunlight ◽  
Free Living ◽  
Lichenized Ascomycetes ◽  
Lobaria Pulmonaria ◽  
Dopa Melanin

AbstractTyrosinases are a widespread family of multicopper oxidase enzymes. In our earlier work, we identified the presence of tyrosinases in lichenized Ascomycetes based on their substratum specificity, sensitivity to inhibitors and molecular mass. Here, we present a more detailed characterization of a tyrosinase from the lichenLobaria pulmonaria. We also compare tyrosinase activity with the activities of laccases and peroxidases, the other redox enzymes present in this species. The importance of tyrosinases in lichen biology was studied by testing their role in melanin synthesis. Laboratory experiments clearly showed that tyrosinases fromL. pulmonariaresemble those from other lichens and in free-living fungi. While the tyrosinases can metabolize the melanin precursor L-DOPA, L-DOPA can also be metabolized by peroxidases and laccases. A field experiment showed that exposing shade-adaptedL. pulmonariato normal solar radiation induces L-DOPA melanin synthesis. Synthesis occurred when lichens were exposed to either direct sunlight, or placed under a wavelength-neutral filter that slightly reduced overall light. In lichens receiving unfiltered sunlight, melanin synthesis was accompanied by increased laccase activity; by contrast, no changes in enzyme activity occurred in lichens placed under the wavelength-neutral filter. Melanization was reduced by placing lichens under filters that removed UV-B, and prevented by filters that removed both UV-A and UV-B. Removing UV-B had no effect on enzyme activity, whereas removing both UV-A and UV-B increased tyrosinase activity. Results from this study indicate that under some conditions laccases may be involved in melanin synthesis, but they provide no evidence for a role for tyrosinases in melanization. Although high tyrosinase activities are widespread in lichens, many questions on the role of this enzyme in lichen biology remain to be answered.

Important role of fungal intracellular laccase for melanin synthesis: purification and characterization of an intracellular laccase from Lentinula edodes fruit bodies

Microbiology ◽  
2003 ◽  
Vol 149 (9) ◽  
pp. 2455-2462 ◽  
Author(s):  
Masaru Nagai ◽  
Maki Kawata ◽  
Hisayuki Watanabe ◽  
Machiko Ogawa ◽  
Kumiko Saito ◽  
...  
Keyword(s):  
Lentinula Edodes ◽  
Veratryl Alcohol ◽  
Size Exclusion ◽  
Melanin Synthesis ◽  
Sds Page ◽  
Diammonium Salt ◽  
Exclusion Chromatography ◽  
Homogeneous Preparation

A laccase (EC 1.10.3.2) was isolated from the fully browned gills of Lentinula edodes fruit bodies. The enzyme was purified to a homogeneous preparation using hydrophobic, cation-exchange and size-exclusion chromatography. SDS-PAGE analysis showed the purified laccase, Lcc 2, to be a monomeric protein of 58·0 kDa. The enzyme had an isoelectric point of around pH 6·9. The optimum pH for enzyme activity was around 3·0 against 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt (ABTS), and it was most active at 40 °C and stable up to 50 °C. The enzyme contained 8·6 % carbohydrate and some copper atoms. The enzyme oxidized ABTS, p-phenylenediamine, pyrogallol, guaiacol, 2,6-dimethoxyphenol, catechol and ferulic acid, but not veratryl alcohol and tyrosine. β-(3,4-Dihydroxyphenyl)alanine (l-DOPA), which was not oxidized by a laccase previously reported from the culture filtrate of L. edodes, was also oxidized by Lcc 2, and the oxidative product of l-dopa was identified as l-DOPA quinone by HPLC analysis. Lcc 2 was able to oxidize phenolic compounds extracted from fresh gills to brown-coloured products, suggesting a role for laccase in melanin synthesis in this strain.

Occurrence of high tyrosinase activity in the non-Peltigeralean lichenDermatocarponminiatum(L.) W. Mann

2012 ◽  
Vol 44 (6) ◽  
pp. 827-832 ◽  
Author(s):  
Richard P. BECKETT ◽  
Farida V. MINIBAYEVA ◽  
Christiane LIERS
Keyword(s):  
Cell Walls ◽  
Redox Cycling ◽  
Tyrosinase Activity ◽  
Melanin Synthesis ◽  
Multicopper Oxidases ◽  
Apparent Absence ◽  
Ph Optimum ◽  
Cellular Location

AbstractIn our earlier work, we demonstrated the presence of the multicopper oxidases tyrosinase and laccase in the cell walls of lichens from thePeltigerales, while these enzymes appeared to be absent in lichens from other orders. Likely roles for tyrosinase in lichens include melanin synthesis, the generation of quinones needed for laccase-mediated redox cycling, and the removal of harmful reactive molecules formed by this cycling. Non-Peltigeralean lichens will not need tyrosinase to detoxify laccase-generated radicals. However, many non-Peltigeralean lichens are often heavily melanized. Apparent absence of tyrosinase activity in these species prompted us to suggest that, in these lichens, melanins are probably synthesized by the polyketide pathway, which does not involve tyrosinase. Here, we surveyed intracellular tyrosinase activity in thallus homogenates from a range of lichens. Results showed that Peltigeralean species generally have much higher activities than species from other orders. However, the non-Peltigeralean lichenDermatocarpon miniatumdisplays significant tyrosinase activity. InD. miniatum, tyrosinase differs from the corresponding enzyme from Peltigeralean lichens with respect to cellular location, substratum specificity, stability and pH optimum. Furthermore, unlike Peltigeralean lichens, inD. miniatumtyrosinase activity increased strongly following the rehydration of dry thalli. These differences are possibly a consequence of the role of tyrosinase in melanin synthesis rather than laccase-mediated redox cycling.

scholarly journals Characterization of a high-affinity membrane-associated ornithine decarboxylase from the free-living nematode Caenorhabditis elegans

1990 ◽  
Vol 270 (3) ◽  
pp. 599-604 ◽  
Author(s):  
J M Schaeffer ◽  
M R Donatelli
Keyword(s):  
Enzyme Activity ◽  
Caenorhabditis Elegans ◽  
Ornithine Decarboxylase ◽  
Decarboxylase Activity ◽  
Nematode Caenorhabditis Elegans ◽  
Free Living ◽  
Affinity Membrane ◽  
Free Living Nematode ◽  
Km Value

Ornithine decarboxylase has been identified and characterized in the free-living nematode Caenorhabditis elegans. Unlike previously described ornithine decarboxylases, the enzyme activity is membrane-associated and remains in the membrane fraction after treatment with high salt, detergents or phosphatidylinositol-specific phospholipase C. Ornithine has an apparent Km value of 2.7 microM for ornithine decarboxylase. The enzyme is competitively inhibited by arginine and lysine with Ki values of 4.0 and 24.4 microM respectively. None of the other naturally occurring amino acids inhibited more than 10% of the enzyme activity at concentrations up to 1 mM. Agmatine, putrescine, spermidine and spermine inhibit ornithine decarboxylase in a non-competitive manner with Ki values of 10, 53.5, 59 and 855 microM respectively. A similar ornithine decarboxylase activity was also identified in membrane preparations from the parasitic nematode Haemonchus contortus.

The role of 6-phosphogluconate dehydrogenase in Rhizobium

1977 ◽  
Vol 23 (9) ◽  
pp. 1293-1298 ◽  
Author(s):  
Kalemani Mulongoy ◽  
Gerald H. Elkan
Keyword(s):  
Enzyme Activity ◽  
Nicotinamide Adenine Dinucleotide ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Pentose Phosphate ◽  
Nicotinamide Adenine ◽  
Phosphogluconate Dehydrogenase ◽  
Ketonic Group ◽  
Fast Growing Rhizobia

A nicotinamide adenine dinucleotide (NAD) linked 6-phosphogluconate (6-PG) dehydrogenase has been detected in Rhizobium. The enzyme activity is similar in both slow- and fast-growing rhizobia. The nicotinamide adenine dinucleotide phosphate (NADP) dependent 6-PG dehydrogenase was detected only in the fast growers and was more than twice as active as the NAD-linked enzyme. Partial characterization of the products of 6-PG oxidation in Rhizobium suggests that the NADP-linked enzyme is the decarboxylating enzyme of the pentose phosphate (PP) pathway (EC 1.1.1.44) whereas a phosphorylated six-carbon compound, containing ketonic group(s), is the product of the oxidation catalyzed by the NAD-linked enzyme.

Digital x-ray mapping of nanometer-size supported bimetallic catalysts

1988 ◽  
Vol 46 ◽  
pp. 530-531
Author(s):  
L. T. Germinario
Keyword(s):  
Background Radiation ◽  
Metal Cluster ◽  
Single Element ◽  
Beam Diameter ◽  
X Rays ◽  
X Ray ◽  
Major Interest ◽  
Induced Changes

Understanding the role of metal cluster composition in determining catalytic selectivity and activity is of major interest in heterogeneous catalysis. The electron microscope is well established as a powerful tool for ultrastructural and compositional characterization of support and catalyst. Because the spatial resolution of x-ray microanalysis is defined by the smallest beam diameter into which the required number of electrons can be focused, the dedicated STEM with FEG is the instrument of choice. The main sources of errors in energy dispersive x-ray analysis (EDS) are: (1) beam-induced changes in specimen composition, (2) specimen drift, (3) instrumental factors which produce background radiation, and (4) basic statistical limitations which result in the detection of a finite number of x-ray photons. Digital beam techniques have been described for supported single-element metal clusters with spatial resolutions of about 10 nm. However, the detection of spurious characteristic x-rays away from catalyst particles produced images requiring several image processing steps.

Sign in / Sign up

Export Citation Format

Share Document