Laccases and their natural mediators: Biotechnological tools for sustainable eco-friendly processes

Laccases are oxidoreductases which oxidize a variety of aromatic compounds using oxygen as the electron acceptor and producing water as by-product. The interest for these old enzymes (first described in 19th century) has progressively increased due to their outstanding biotechnological applicability. The presence of redox mediators is required for a number of biotechnological applications, providing the oxidation of complex substrates not oxidized by the enzyme alone. The efficiency of laccase–mediator systems to degrade recalcitrant compounds has been demonstrated, but still the high cost and possible toxicity of artificial mediators hamper their application at the industrial scale. Here, we present a general outlook of how alternative mediators can change this tendency. We focus on phenolic compounds related to lignin polymer that promotes the in vitro transformation of recalcitrant non-phenolic structures by laccase and are seemingly the natural mediators of laccases. The use of eco-friendly mediators easily available from lignocellulose, could contribute to the industrial implementation of laccases and the development of the 21th century biorefineries.     

Physiological performance of silver-fir (Abies alba Mill.) populations under contrasting climates near the south-western distribution limit of the species

Silver-fir (Abies alba Mill.) populations located at the south side of the main Pyrenean axis and Pre-Pyrenees constitute the south-western distribution limit of the species and, therefore, may be responding more noticeably to climate change than those in the centre of the range. The increasing aridity detected in the Pyrenees during the 20th century should affect more negatively the physiological performance of the southernmost silver-fir stand growing under more xeric conditions in comparison with stands growing within the main distribution area under mesic conditions. To evaluate the climatic influence on the performance of silver fir near its distribution limit, we studied several physiological and growth variables in shoots and needles from two silver fir populations located in nearby but climatically contrasting sites: Paco Ezpela (site E) and Gamueta (site G). Site E showed a stronger Mediterranean influence than site G, i.e. the former site was characterized by higher temperatures and leaf-to-air water vapour pressure difference and lower precipitation in summer than the latter site. Silver firs from site E showed lower values of primary and secondary growth, needle length, stomatal conductance, net photosynthesis and photosystem II (PSII) efficiency than individuals from site G. The reduction in net photosynthesis could be ascribed to a low CO₂ availability and to a lower PSII efficiency. We conclude that the physiological differences found between both sites were caused by the more xeric conditions of site E as compared with the more mesic environment in site G. The predicted increase of severe droughts in the southern Pyrenees might cause a decrease in photosynthesis and growth in those silver-fir stands located near the ecological limit of the species.     

Solution versus solid-phase cyclization strategies for large sidechain lactam-bridged peptides: A comparative study

A 22-residue peptide with a sidechain lactam bridge involving 18 residues (60-atom cycle) has been synthesized. Three different protection schemes using Fmoc/^tBu/cyclohexyl, Fmoc/^tBu/allyl or Boc/Bzl/ fluorenylmethyl protecting group combinations have been explored for the solid phase of the linear precursors, which have been subsequently cyclized in solution or in the solid phase. Cyclization yields in solution have been consistently better than on solid phase; however, the solid-phase strategy requires fewer purification steps and therefore global yields are comparable.     

Engineering and Applications of fungal laccases for organic synthesis

Laccases are multi-copper containing oxidases (EC 1.10.3.2), widely distributed in fungi, higher plants and bacteria. Laccase catalyses the oxidation of phenols, polyphenols and anilines by one-electron abstraction, with the concomitant reduction of oxygen to water in a four-electron transfer process. In the presence of small redox mediators, laccase offers a broader repertory of oxidations including non-phenolic substrates. Hence, fungal laccases are considered as ideal green catalysts of great biotechnological impact due to their few requirements (they only require air, and they produce water as the only by-product) and their broad substrate specificity, including direct bioelectrocatalysis.     

Fast replenishment of initial carbon stores after defoliation by the pine processionary moth and its relationship to the re-growth ability of trees

Defoliation by herbivores may alter the source:sink balance of trees leading to transient decreases in carbon (C) stores. When C stores are replenished concurrently with re-growth both processes may compete, store formation proceeding at the expenses of growth. However, the interactions between both processes are not fully understood. We investigated the effects of defoliation by the pine processionary moth (PPM, Thaumetopoea pityocampa Dennis and Schiff.) on the non-structural carbohydrate (NSC) and nitrogen (N) stores and the growth of Pinus nigra Arnold trees. Short-term effects were evaluated immediately after a PPM outbreak and at the end of the first growing season in trees suffering a range of defoliation damage. Long-term effects were explored by a 17-year-long PPM defoliation experiment, with 11?years of repeated defoliation treatments followed by 6?years of recovery. Defoliation by PPM was followed by transient NSC decreases, but trees were able to exceed initial NSC pools and compensate growth in just one growing season. Such recovery was linked to increased foliage N. Repeated severe defoliations decreased growth and survival of trees in the long-term, but trees increased starch allocation to stems. Defoliation led to an accumulation of C storage compounds in P. nigra trees irrespective of their ability to re-grow. In trees included in the short-term experiment, the accumulation of stores proceeded concurrently with re-growth. However, the repeated severe defoliations included in our long-term experiment impaired the growth of trees, surplus C being accumulated as stores. These results indicate that, growth declines in pines defoliated by PPM are not due to C (source) limitation but may respond to the reduced sink strength of growing meristems due to defoliation, and thus, a decrease in C allocation to growth.     

Studies on antigenic variability of C strains of foot-and-mouth disease virus by means of synthetic peptides and monoclonal antibodies.

Peptides representing the sequence of the immunodominant loop of foot-and-mouth disease virus strain C-S8 (YTASARGDLAHLTTTHARHLP, residues 136-156 of VP1) and of several variant viruses have been prepared by solid phase methods. In addition, five peptides with single-residue replacements at Leu147 (Ile, Nle, Val, Ala, Gly) have been synthesized. Tosyl and dinitrophenyl protections for histidine have been compared, the latter being found to give better synthetic products. The peptides have been tested in an immunodot assay against a panel of monoclonal antibodies directed towards the VP1 loop. Immunochemical results are discussed on the basis of the mobility of the region reproduced by the peptides and the nature of the side chain of residue 147.     

A Circadian Rhythm in the Activity of Superoxide Dismutase in the Photosynthetic Alga Gonyaulax polyedra

The activity of superoxide dismutase in cell-free extracts of Gonyaulax made at different times of day and night was found to be three to four times higher during the day. This rhythm continued in cells kept in constant light, indicating that the regulation can be attributed to the cellular circadian clock.