Decolorization of effluent from the bagasse-based pulp mills by white-rot fungus,Schizophyllum commune

Summary The effluent from bagasse-based pulp and paper mills can be decolorized with the white-rot fungus Schizophyllum commune. The influence of pH, nutrients and aeration on the decolorizing efficiency of this fungus has been determined. It was found that it could not degrade lignin unless a more easily metabolized carbon source was made available simultaneously. The addition of carbon and nitrogen not only improved the decolorizing efficiency of the fungus, but also resulted in reduction of the biological oxygen demand (BOD) and chemical oxygen demand (COD) of effluent. A 2-day incubation period was sufficient for lignin breakdown by S. commune. The efficiency of treatment of effluent with this fungus was highest at pH 4–5 and was further improved by intermittent aeration.     

Nutritional study on <Emphasis Type="Italic">Embellisia astragali</Emphasis>, a fungal pathogen of milk vetch (<Emphasis Type="Italic">Astragalus adsurgens</Emphasis>)

Embellisia astragali is a strong, virulent pathogen that develops within milk vetch (Astragalus adsurgens). In order to determine nutrient requirements, the fungus was cultured on 9 carbon sources, 9 nitrogen sources, and 13 growth media in the dark at 25°C. Growth rates and sporulation capacity were measured after 4 and 12 weeks. All carbon sources supported growth, but only soluble starch, inulin, and dextrose supported sporulation. In general, better growth was obtained on disaccharides and polysaccharides than on monosaccharides. Compared with no growth on NH₄ ⁺-N and urea, the fungus grew little on all NO₃ ⁻-N, amino-N, and other organic-N such as peptone. There was no sporulation or very sparse conidia on almost all nitrogen sources with supplied dextrose or soluble starch as sole carbon source. The better growth and sporulation on most of the semidefined media than on defined media indicates that some components in plant or animal material may be vital to the fungus. Sporulation was positively correlated with growth rate in N source experiment at 12 weeks and in growth media experiment at 4 and 12 weeks. The fungus favors grow within agar with growth rate less than 1.18 mm day⁻¹.     

Carbon and nitrogen requirements of Fusarium oxysporum causing cotton wilt

Summary The present work was carried out to study the nutritional requirements of the cotton wilt-inducing fungus, i.e.Fusarium oxysporum on a synthetic liquid medium with regard to the carbon and nitrogen sources at varying concentrations in terms of the average mycelial dry weights.The optimum carbon requirements of the fungus ranged from 7000–8000 p.p.m. irrespective of the carbon source used in experiment. Carbon utilization was best on sucrose followed by maltose, starch, glucose, fructose and cellulose successively.The optimum nitrogen requirements of the fungus were 300 p.p.m. of nitrogen in the medium; nitrogen utilization was best on using nitrate-nitrogen followed by glycine, glutamic acid, ammonium nitrate, asparagine and ammonium sulphate.Maximum growth of the fungus took place on media containing a C/N ratio ranging between 22.8 and 25.7.Colour formation is correlated with varying either source or concentration of nitrogen and not carbon.     

Fruiting-body formation, cultivation properties, and host specificity of a fungicolous fungus, Asterophora lycoperdoides

We report the fruiting-body formation and cultivation properties of Asterophora lycoperdoides, a fungicolous fungus. Asterophora lycoperdoides formed fruiting bodies on potato dextrose agar medium in approximately 1 week, although this fungus shows high host specificity to Russula nigricans in nature. Optimal temperature of mycelial growth and fruiting-body formation was 25°C. Mannitol or soluble starch was preferably used as a carbon source, and amino nitrogen was preferably used as the nitrogen source. For a better understanding of the relationship between A. lycoperdoides and R. nigricans, we cultivated A. lycoperdoides on media supplemented with freeze-dried fruiting bodies of various fungi. The germination rate was approximately 2.5 times higher on the medium containing freeze-dried R. nigricans than that on the PDA medium. The mycelia extended most rapidly in the presence of R. nigricans. Furthermore, the stipe length of its fruiting body was the longest on the medium containing R. nigricans. These results indicated that A. lycoperdoides can grow faster by utilizing certain substances that are abundantly contained in R. nigricans, such as mannitol, or by utilizing R. nigricans itself. It is considered that the constituents of R. nigricans might contribute to the host specificity of A. lycoperdoides.     

Evaluation of the effect of process variables on the fatty acid profile of single cell oil produced by Mortierella using solid-state fermentation

This article reviews some of the aspects of single cell oil (SCO) production using solid-state fermentation (SSF) by fungi of the genus Mortierella. This article provides an overview of the advantages of SSF for SCO formation by the aforementioned fungus and demonstrates that the content of the polyunsaturated fatty acids (PUFA) depend on the type of fermentation media and culture conditions. Process variables that influence lipid accumulation by Mortierella spp. and the profile of the fatty acids are discussed, including incubation temperature, time, aeration, growth phase of the mycelium, particle size of the substrate, carbon to nitrogen ratio, initial moisture content and pH as well as supplementation of the substrate with nitrogen and oil. Finally, the article highlights future research trends for the scaled-up production of PUFAs in SSF.     

Optimization of process parameters for improved production of bioactive metabolite by a novel endophytic fungus <Emphasis Type="Italic">Fusarium</Emphasis> sp. DF2 isolated from <Emphasis Type="Italic">Taxus wallichiana</Emphasis> of North East India

An endophytic fungus having antifungal and antibacterial properties was isolated from Taxus wallichiana of Arunachal Pradesh, India. On the basis of morphological and molecular characteristics, the fungus was identified as Fusarium sp. and designated as DF2. The fungus was optimized for growth and maximum production of the antimicrobial agent. Media with 5% leaf extract (w/v) supplemented with 0.1% dextrose as carbon and yeast extract as nitrogen source favored the growth with temperature optimum 25 ± 2°C and pH 6. Incubation period of 10 days was observed optimum for growth and production of antimicrobial agent. Phenylalanine and dextrose enriched basal medium promoted the antimicrobial agent production, whereas methionine amended in combination with glucose promoted higher biomass accumulation. The TLC purified active compound with UV λ-max 270 nm in ethyl acetate has got the lowest minimum inhibitory concentration (MIC) against Bacillus subtilis, Staphylococcus aureus and Escherichia coli and highest against Pseudomonas aeruginosa.     

Oil Cakes as Media for Growing Catenaria anguillulae Sorokin, a Facultative Endoparasite of Nematodes

Summary Growth, morphology, visibility of sporangia and colony colour of 10 isolates of Catenaria anguillulae were compared on six media: linseed oil-cake agar, mustard oil-cake agar, neem oil-cake agar, beef extract agar, Emerson agar and YPSS agar with a view to selecting the best growth medium. In general, maximum radial growth of most of the isolates was recorded on linseed oil-cake agar medium, whereas neem oil-cake agar medium supported least growth of all the isolates of C. anguillulae. Linseed oil-cake agar medium also maintained the typical characters of the fungus and clear visibility of morphological details.     

Production of <Emphasis Type="Italic">Anti-Helicobacter pylori</Emphasis> metabolite by the lichen-Forming fungus <Emphasis Type="Italic">Nephromopsis pallescens</Emphasis>

The present study was conducted to evaluate the antibacterial activity of lichen-forming fungi (LFF) against Helicobacter pylori, and to optimize the culture conditions of LFF for maximum production of natural antibiotics against H. pylori. To accomplish this, a screening assay was first conducted among 19 species of LFF. The extract of Nephromopsis pallescens (KOLRI-040516) exhibited the strongest anti-ff. pylori activity. Bioautograghic TLC and HPLC analysis identified usnic acid as the main antibacterial substance produced by JV. pallescens. The growth of JV. pallescens and production of antibacterial substances produced by the fungus were then investigated under several culture conditions including the culture media, initial medium pHs, incubation temperatures, and the degree of aeration. The results indicated that culture in MY medium with an initial pH of 6.0, a temperature of 15°C and a low degree of aeration supported the largest usnic acid production of the fungus (16.4 ug usnic acid/g dry biomass). Especially, aeration was found to be an important factor that affect both growth and usnic acid production of N. pallescens.     

An inexpensive medium for production of arachidonic acid by <Emphasis Type="Italic">Mortierella alpina</Emphasis>

The production of arachidonic acid was studied in the fungus Mortierella alpina using an inexpensive medium. Glucose derived from maize starch hydrolysate was the sole carbon source and defatted soybean meal and sodium nitrate were the nitrogen sources. Optimal arachidonic acid yield (1.47 g l^-1) was observed at a glucose concentration of 100 g l^-1. Various treatments of defatted soybean meal to extract soluble nitrogen nutrients were evaluated. Alkali extract was the most effective for arachidonic acid production. A mixture of soybean alkali-extract protein and sodium nitrate was an excellent nitrogen source for fungal growth, lipid accumulation, and arachidonic acid production. A maximum yield of 1.87 g arachidonic acid l^-1 was obtained with a soybean protein concentration of 4.6 g l^-1 and a sodium nitrate concentration of 2.3 g l^-1. Electronic Publication     

The influence of cultural medium composition on the proteolytic enzyme secretion of fungus <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

It was shown that change of medium growth composition of photopathogenic fungus Rhizoctonia solani Kühn, especially accessible sources of nutrition, leads to change of both quantity of produced proteinases and their action specificity. The mineral source of nitrogen suppressed the fungus proteinase secretion on cultivatiin medium containing potato thermostable proteins but an organic source of nitrogen accelerated mycelium growth and increased proteinase secretion. On the basis of an analysis of a fungus extracellular proteinase substrate-specificity, it is established that the presence of thermostable proteins of a potato in the cultural liquid induces the secretion of trypsin-like proteinases mainly, and the addition of yeast extract to this growth medium induces the secretion of subtilisin-like ones, thus suppressing the trypsin-like enzymes production. This fact can indicate that mycelium of fungus R. solani loses pathogenic properties and becomes saprophytes when the growth medium was enriched by an organic source of nitrogen.     

Experimental Studies on the Physiology of the Phycobiont and Mycobiont Ramalina ecklonii By

The lichenized fungus and alga of the fruticose lichen Ramalini ecklonii were isolated into pure cultures. The ascospores of the fungus failed to germinate in less than five weeks incubation in spite of the use of a variety of cultural conditions. The fungus showed a considerable increase in growth on malt extract agar. Both organisms showed a marked tolerance for high concentrations of glucose although growth was quantitatively reduced. The fungus was able to use a variety of carbon and nitrogen sources as well as an extract of algal cells. Cultivation in the absence of biotin and thiamine failed to yield significant amounts of growth. The alga yielded 27 mg of dry weight after three weeks in a synthetic medium under low light intensities. The alga could be grown in satisfactory amounts on CO₂ and inorganic salts with moderate light intensities. Experiments using ¹⁴CO₂ showed the fungus able to incorporate the extra-cellular and intra-cellular products of algal metabolism. The rate of incorporation of extra-cellular products was inhibited by high concentrations of biotin and thiamine. The alga assimilated ^l4CO₂ which was retained by the cells over a period of 14 days, at which time 78 per cent of the activity was insoluble in 80 per cent ethanol. An extract of the fungus labelled with ¹⁴C glucose was partially taken up by the alga and 50 per cent of the label was insoluble in 80 per cent after three days incubation in the light. No lichen acids were found in either the fungal cultures or the algal cultures although large amounts (e.g. 2 liters) of material were extracted and chromatographed. Usnic acid was produced by the intact lichen thallus.     

PHB accumulation in Nostoc muscorum under different carbon stress situations

Use of algae for intracellular poly-β-hydroxybutyrate (PHB) accumulation for bioplastic production offers an opportunity in economic efficiency by reduced costs. The cyanobacterium Nostoc muscorum is a PHB accumulator which presents a great potential as raw material supplier because of short generation cycles. Here, we examined a range of experimental conditions including different growth conditions of phosphate-starved cells with the addition of external carbon sources. The highest, absolute PHB accumulation was measured in a phosphate-starved medium with 1% (w/w) glucose and 1% (w/w) acetate. PHB accumulated inside algae cells. After 23 days of growth in phosphate-starved medium, 1 L of culture contained up to 145.1 mg PHB. The highest PHB accumulation based on the cell dry weight was in an experiment with aeration and CO₂ addition. The intracellular level of PHB was up to 21.5% cell dry weight after 8 days.     

Effect of iron and aeration on superoxide dismutase and catalase activity of PHB-producing Azotobacter chroococcum

The effect of aeration level and iron concentration on Azotobacter chroococcum 23 growth, PHB accumulation and antioxidative enzyme activities was investigated in shake flask experiments. Biomass yield and carbon source conversation coefficients increased in the presence of iron in the growth medium and under decreased aeration. The highest biomass production was observed for the culture grown in a medium with 36 μM of initial iron concentration and moderate aeration level. The highest PHB accumulation level (70–72% from cell dry weight) under our experimental conditions was observed at decreased aeration in the growth medium with 180 μM of initial iron concentration. Results obtained prove that both aeration level and iron supply have a marked influence on the activity of SOD and catalase. Bearing in mind the necessity of iron for the synthesis of both enzymes, only catalase showed a specific dependence on the intracellular iron accumulation level.     

Bacterial community dynamics in aerated cow manure slurry at different aeration intensities

Aims: This study aimed to characterize microbial community dynamics in aerated cow manure slurry at different aeration intensities. Methods and Results: Batch aerobic treatments were set up in 5‐l jar fermentor, each containing 3 l of manure slurry; the slurries were subjected to low, medium and high (50, 150 and 250 ml min^?1, respectively) aeration for 9 days. Microbial community composition was determined using terminal restriction fragment length polymorphism and a clone library targeting 16S rRNA genes. High and medium aeration accelerated organic carbon degradation in parallel with the degree of aeration intensity; however, 90% of the initial total organic carbon was retained during low‐aeration treatment. During the active stages of organic carbon decomposition, clones belonging to the class Bacilli accumulated. Moreover, Bacilli accumulation occurred earlier under high aeration than under medium aeration. Conclusions: Organic matter degradation was mainly governed by a common microbial assemblage consisting of many lineages belonging to the class Bacilli. The timing of community development differed depending on aeration intensity. Significance and Impact of the Study: This study reports on changes in several environmentally important parameters and the principal microbial assemblage during the pollution‐reducing phase of cattle manure aeration treatment.     

Organic nitrogen of tomato waste hydrolysate enhances glucose uptake and lipid accumulation in Cunninghamella echinulata

Aims: To investigate the effect of organic nitrogen on lipogenesis during growth of Cunninghamella echinulata on tomato waste hydrolysate (TWH) media. Methods and Results: Cunninghamella echinulata grown on a TWH medium rapidly took up glucose and produced large amounts of lipids. However, when some quantities of the organic nitrogen were removed from TWH (by acid followed by alkaline precipitation of proteins) the uptake of glucose was dramatically reduced and large quantities of fungal biomass having low lipid content were produced. Nevertheless, when glycerol was used as carbon source instead of glucose, the uptake rate as well as the biomass production and the lipid accumulation processes were unaffected by the TWH organic nitrogen removal. Finally, when the fungus was grown on a glucose supplemented TWH medium that contained no assimilable organic nitrogen (after further precipitation of proteins with methanol), the produced biomass contained non-negligible quantities of lipids, although glucose uptake remained low. Lipid analysis showed that the produced lipids comprised mainly of neutral lipids, which were preferentially consumed during lipid turnover. Lipid production on the original TWH medium having glucose as carbon source was 0·48 g of lipid per gram of dry biomass, corresponding to 8·7 g of lipid per litre of growth medium. The produced lipids contained 11·7%γ-linolenic acid (GLA), hence the GLA yield was more than 1 g l⁻¹. Conclusions: Organic nitrogen compounds found in TWH favour glucose (but not glycerol) uptake and lipid accumulation in C. echinulata. Significance and Impact of the Study: Agro-industrial wastes containing organic nitrogen, such as tomato waste, are produced in vast amounts causing severe environmental problems. These wastes could be used as fermentation feedstock to produce microbial lipids.     

Semi-defined Medium for in vitro Cultivation of the Fastidious Insect Pathogenic Fungus Cordyceps unilateralis

Cordyceps unilateralis is a fastidious fungal pathogen affecting ants. Up to now, only the complex and expensive Grace’s insect cell culture medium has been used for in vitro cultivation (as blastospores and mycelium) of this fungus. To obtain an inexpensive and less complicated medium, the effects of carbon and nitrogen sources, salt solution and carbon-to-nitrogen (C:N) ratio on the growth of this fungus were examined. Glucose was the most important factor for blastospore formation, and yeast extract could be used as a nitrogen source for blastospore formation and mycelial growth. A suitable C:N ratio (glucose: yeast extract) was 33.3:1. As a result, a new semi-defined medium was achieved, composed of 26.68 g L⁻¹ glucose, 3.3 g L⁻¹ yeast extract and salt solution. This medium supported blastospore formation and mycelial growth of all tested C. unilateralis isolates.     

Evaluation of a new fungus <Emphasis Type="Italic">Ceriporia lacerate</Emphasis> strain P2—its ability to decolorize Alizarin Red and Methyl Orange

A new fungus Ceriporia lacerate P2 which belongs to family Polyporaceae was evaluated for its ability to decolorize two different dyes Alizarin Red and Methyl Orange. Different parameters such as incubation time, pH, carbon source, nitrogen source and carbon/nitrogen regimes were used to find out the optimum medium for Ceriporia lacerate P2 on its ability of decolorization. The results show that the fungus had different ability to decolorize the two tested dyes. For Alizarin Red, the most suitable medium was at pH 3 and the best carbon and nitrogen source were sucrose and ammonium nitrate. While for Methyl Orange, the optimum medium was at pH 7–9 and the best carbon and nitrogen source were sucrose and urea.     

Effect of aeration and agitation on the biosynthetic activity of diffusely growingAspergillus niger

A considerable lowering of aeration demands occurs during diffuse growth of citric acid-producingAspergillus niger in a submerged cultivation. However, the diffuse culture poses stricter demands on the type of aeration and agitation. The impeller frequency affects considerably the morphology of the producer fungus and the accumulation of citric acid. The effect of impeller frequency on the distribution of air in the medium and on the amount of air fed into the diffuse culture is less important.     

<Emphasis Type="Italic">Rhodococcus</Emphasis> sp. Q5, a novel agarolytic bacterium isolated from printing and dyeing wastewater

An agar-degrading bacterium, Rhodococcus sp. Q5, was isolated from printing and dyeing wastewater using a mineral salts agar plate containing agar as the sole carbon source. The bacterium grew from pH 4.0 to 9.0, from 15 to 35°C, and in NaCl concentrations of 0–5 %; optimal values were pH 6.0, 30°C, and 1 % NaCl. Maximal agarase production was observed at pH 6.0 and 30°C. The bacterium did not require NaCl for growth or agarase production. The agarase secreted by Q5 was inducible by agar and was repressed by all simple sugars tested except lactose. Strain Q5 could hydrolyze starch but not cellulose or carboxymethyl cellulose. Agarase activity could also be detected in the medium when lactose or starch was the sole source of carbon and energy. Strain Q5 could grow in nitrogen-free mineral media; an organic nitrogen source was more effective than inorganic carbon sources for growth and agarase production. Addition of more organic nitrogen (peptone) to the medium corresponded with reduced agarase activity.     

Improvement of Bacillus thuringiensis Bacteriocin Production Through Culture Conditions Optimization

Abstract

BUPM4 is a Bacillus thuringiensis subsp. kurstaki strain, isolated from Tunisian soil, producing an original bacteriocin named Bacthuricin F4. The optimization of the latter production conditions was carried out under several physicochemical conditions. It was found that the highest bacteriocin activity was reached at low aeration while bacteriocin synthesis yields were strongly reduced at higher ones. A balance between growth and bacteriocin synthesis, both highly dependent on aeration, was taken into account for the overproduction of bacteriocin. Both glucose and glycerol were shown to be necessary for Bacthuricin F4 maximal synthesis. In addition, the optimal carbon/nitrogen ratio for bacteriocin production is 9. In such optimal conditions, more than 4-fold greater bacteriocin production was obtained than when using TSB medium.