Effects of three esca-associated fungi on Vitis vinifera L.: III. Enzymes produced by the pathogens and their role in fungus-to-plant or in fungus-to-fungus interactions

When the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme) were grown in liquid stationary cultures, it was seen that they were able to live in media containing resveratrol (RES) or tannic acid (TA) as the sole carbon source and that the fungi were able to convert both compounds. Particular attention is paid here to detecting RES and TA conversion. Pch, Tmi and Fme were partially inhibited by RES or TA. Pch, Tmi and Fme produced extracellular tannase, laccase and peroxidase enzymes in liquid or agarized cultures, whether glucose was present or not. When colonies of Pch, Tmi and Fme were confronted, they showed spatially and temporally heterogeneous patterns of laccase and peroxidase activity. The results indicate the non-synergistic, competitive association of Pch and Tmi and the inhibition of Fme growth. Muconic acid, a well-known intermediate in a large number of lignin and phenol oxidative processes, can partly or completely inhibit the lignolytic agent Fme, but is tolerated by Pch and Tmi. An explanation for wood pigmentation patterns by Pch, Tmi and Fme is given.     

Treatment of wheat straw using tannase and white-rot fungus to improve feed utilization by ruminants

Background

Current research to enrich cattle feed has primarily focused on treatment using white rot fungi, while there are scarce reports using the enzyme tannase, which is discussed only in reviews or in the form of a hypothesis. In this context, the aim of the present study was to evaluate the effect of tannase on wheat straw (WS) and also the effect of lyophilized tannase at concentrations of 0.1%, 0.2%, and 0.3% (w/w) on WS followed by fermentation with Ganoderma sp. for 10 d and compared in relation to biochemical parameters, crude protein (CP) content, and nutritional value by calculating the C/N ratio in order to improve the nutritional value of cattle feed.

Results

Penicillium charlesii, a tannase-producing microorganism, produced 61.4 IU/mL of tannase in 54 h when 2% (w/v) tannic acid (TA) was initially used as a substrate in medium containing (% w/v) sucrose (1.0), NaNO₃ (1.0), and MgSO₄ (0.08 pH, 5.0) in a 300-L fermentor (working volume 220 L), and concomitantly fed with 1.0% (w/v) TA after 24 h. The yield of partially purified and lyophilized tannase was 5.8 IU/mg. The tannin-free myco-straw at 0.1% (w/w) tannase showed 37.8% (w/w) lignin degradation with only a 20.4% (w/w) decrease in cellulose content and the in vitro feed digestibility was 32.2%. An increase in CP content (up to 1.28-fold) along with a lower C/N ratio of 25.0%, as compared to myco-straw, was obtained.

Conclusions

The use of tannin-free myco-straw has potential to improve the nutritional content of cattle feed. This biological treatment process was safe, eco-friendly, easy to perform, and was less expensive as compared to other treatment methods.     

Polyphenol oxidase, tannase and proteolytic activity in relation to tannin concentration in the soil organic horizon under silver birch and Norway spruce

Our aim was to compare enzyme activities (tannase, polyphenol oxidase and protease) with concentrations of tannins and their ability to precipitate proteins in the litter layer and the humus layer under silver birch (Betula pendula Roth.) and Norway spruce (Picea abies L.). We also estimated the influence of these enzymes on protein-tannin complexes and the influence of tannins on proteolytic activity. The study site was a tree species experiment in Eno, middle-eastern Finland, having three replicated plots dominated by 42-year-old silver birch and Norway spruce. Our hypotheses were (1) tree species and soil layer have an influence on tannin concentrations and enzyme activities, (2) that tannin and protein concentrations in soil organic horizon are positively correlated with enzyme activities and (3) that the enzymes studied have the ability to degrade tannin-protein complexes and that tannins can inhibit proteolytic activity. Concentrations of total tannins and hydrolysable tannins, and tannase and proteolytic activities were higher in the humus layer than in the litter layer. In general the highest values of concentrations of total tannins and hydrolysable tannins and enzyme activities were obtained for the birch humus layer, but the concentrations of condensed tannins and proteins were highest in the litter layer and under spruce. A strong correlation between substrate concentration and enzyme activity was found between hydrolysable tannins and tannase activity. Polyphenol oxidase showed similar activities in both layers. To study the influence of enzymes on protein-tannin complex we synthesized such complexes using bovine serum albumin and either condensed tannins from silver birch and Norway spruce needles or a hydrolysable tannin, tannic acid. Studies with commercial enzymes and enzymes extracted from the soil showed some decrease in tannin concentration of the tannin-protein complex over time, but surprisingly, only a negligible decrease in protein concentration. Complexes of protein with condensed tannins were more recalcitrant than tannic acid-protein complexes. Tannins, depending on the concentration and chemical structure, tended to inhibit proteolytic activity. Our results indicate that protein-tannin complexes are relatively recalcitrant since the enzymes studied here do not effectively release protein from the complexes. Also proteolytic activity and the concentration of extractable proteins seem to be low in soil. However, tannin-degrading enzymes showed high activities.     

Extractives of <Emphasis Type="Italic">Quercus crispula</Emphasis> sapwood infected by the pathogenic fungi <Emphasis Type="Italic">Raffaelea quercivora</Emphasis> I: comparison of sapwood extractives from noninfected and infected samples

The extracts of Quercus crispula infected by the ambrosia fungus, Raffaelea quercivora, were investigated. Phenol and tannin analyses indicated that normal sapwood (NS) contained a considerable amount of hydrolysable tannins, while infected colored sapwood (IS) contained less hydrolysable tannins and more phenols than NS. In treating pentagalloyl glucose (PGG), which is a model compound of hydrolysable tannins, with a culture medium of R. quercivora, PGG was rapidly hydrolyzed to produce gallic acid. The resulting gallic acid decreased in concentration over the subsequent cultivation period eventually disappeared. Measuring tannase and laccase activities of the culture medium of R. quercivora, tannase activity increased gradually from the beginning, while laccase activity increased rapidly at 5 days of incubation and disappeared at 8 days. An oxidative product from gallic acid treated with laccase was isolated by preparative high performance liquid chromatography, and was identified as purprogallincarboxylic acid (PGCA) by nuclear magnetic resonance spectroscopy and electron-impact mass spectrometry. PGCA was present in a 70% aqueous acetone extract of IS, and showed slight growth inhibition against R. quercivora. Part of this study was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, Japan, 2007     

单宁酶的研究进展

详细介绍了单宁酶的来源、生产、提取、纯化及其物理化学和生物学性质,并概述了它的固定化技术。     

Treatment of wheat straw using tannase and white-rot fungus to improve feed utilization by ruminants

Background： Current research to enrich cattle feed has primarily focused on treatment using white rot fungi, while there are scarce reports using the enzyme tannase, which is discussed only in reviews or in the form of a hypothesis. In this context, the aim of the present study was to evaluate the effect of tannase on wheat straw （WS） and also the effect of lyophilized tannase at concentrations of 0.1%, 0.2%, and 0.3% （w/w） on WS followed by fermentation with Ganodermo sp. for 10 d and compared in relation to biochemical parameters, crude protein （CP） content, and nutritional value by calculating the C/N ratio in order to improve the nutritional value of cattle feed. Results： Penicillium charlesii, a tannase-producing microorganism, produced 61.4 IU/mL of tannase in 54 h when 2% （w/v） tannic acid （TA） was initially used as a substrate in medium containing （% w/v） sucrose （1.0）, NaNO3 （1.0）, and MgSO4 （0.08 pH, 5.0） in a 300-L fermentor （working volume 220 L）, and concomitantly fed with 1.0% （w/v） TA after 24 h. The yield of partially purified and lyophilized tannase was 5.8 IU/mg. The tannin-free myco-straw at 0.1% （w/w） tannase showed 37.8% （w/w） lignin degradation with only a 20.4% （w/w） decrease in cellulose content and the in vitro feed digestibility was 32.2%. An increase in CP content （up to 1.28-fold） along with a lower C/N ratio of 25.0%, as compared to myco-straw, was obtained. Conclusions： The use of tannin-free myco-straw has potential to improve the nutritional content of cattle feed. This biological treatment process was safe, eco-friendly, easy to perform, and was less expensive as compared to other treatment methods.