Effect of Trigonella foenum Extract and ZiO2 Nanoparticles on Some Pathogenic Fungi and Bacteria

Aim: The current venture, were made to evaluate the inhibitory effect of Trigonella foenum seed Extract and ZiO2 Nanoparticles on some selected species of Fungi and Bacteria. Materials and Methods: two bacterial species included Pseudomonas aeruginosa and Staphylococcus aureus and three fungal species which is Cryptococcus neoformans, Candidda albicans and Chaetomium were used to evaluate the antibacterial activity of Trigonella foenum Extract and ZiO2 Nanoparticles. Results: This study showed that the Zirconium Oxide (ZiO2) nanoparticles have antifungal and antibacterial activities on the isolates of Cryptococcus neoformans, Candida alicans and Staphylococcus aureus, respectively. While the antimicrobial activity of Zirconium Oxide nanoparticles on the Chaetomium and Pseudomonas aeruginosa was negative. All tested fungi and bacterial isolates were found to be sensitive to Trigonella foenum seed extract, the results of the compination of the ZiO2 Nanoparticle and the Trigonella foenum seed extract were poisitive for all tested fungi isolates and bacterial isolates. The XRD analysis was done for Zirconium Oxide nanoparticles and the result showed that the biocrystallization on the surface of the Zirconium Oxide manoparticles. The average partides size was about (29.8) nm. Conclusions: This investigation conclude that the use of Trigonella foenum seed Extract has the effect of killing all bacteria and fungi under study, result indicate the Trigonella foenun seed Extract best antibacterial efficacy than the ZiO2 together (AU)

Isolation,screening and identification of endophytic fungi and detection of its antifungal effects against Alternaria panax / 中国中药杂志

To obtain biocontrol fungus for Alternaria panax,the antifungal effects of one strain of endophytic fungi isolated from leaves of healthy ginseng were screened and evaluated by using dual-culture method,and the taxonomic assignment of the screened strain was identified based on the morphological characters and ITS sequence analysis. The results of dual-culture showed that one of the endophytes marked as FS-01 had good antifungal effects and the inhibitory rates of FS-01 strain to A. panax was( 60. 21±0. 12) %.The hyphae junction of the both strains,A. panax dissolved,broke and winded,while the hyphae of FS-01 strain remained normal. The inhibitory rates of non-sterilized FS-01 strain fermentation liqud was( 13. 94±0. 21) %. Strain FS-01 identified as Chaetomium globosum.

Isolation, Identification and Enzymatic Activity of Halotolerant and Halophilic Fungi from the Great Sebkha of Oran in Northwestern of Algeria

The Great Sebkha of Oran is a closed depression located in northwestern of Algeria. Despite the ranking of this sebkha among the wetlands of global importance by Ramsar Convention in 2002, no studies on the fungal community in this area have been carried out. In our study, samples were collected from two different regions. The first region is characterized by halophilic vegetation and cereal crops and the second by a total absence of vegetation. The isolated strains were identified morphologically then by molecular analysis. The biotechnological interest of the strains was evaluated by testing their ability to grow at different concentration of NaCl and to produce extracellular enzymes (i.e., lipase, amylase, protease, and cellulase) on solid medium. The results showed that the soil of sebkha is alkaline, with the exception of the soil of cereal crops that is neutral, and extremely saline. In this work, the species Gymnoascus halophilus, Trichoderma gamsii, the two phytopathogenic fungi, Fusarium brachygibbosum and Penicillium allii, and the teleomorphic form of P. longicatenatum observed for the first time in this species, were isolated for the first time in Algeria. The halotolerance test revealed that the majority of the isolated are halotolerant. Wallemia sp. and two strains of G. halophilus are the only obligate halophilic strains. All strains are capable to secrete at least one of the four tested enzymes. The most interesting species presenting the highest enzymatic index were Aspergillus sp. strain A4, Chaetomium sp. strain H1, P. vinaceum, G. halophilus, Wallemia sp. and Ustilago cynodontis.

New production process of the antifungal chaetoglobosin A using cornstalks

Abstract Chaetoglobosin A is an antibacterial compound produced by Chaetomium globosum, with potential application as a biopesticide and cancer treatment drug. The aim of this study was to evaluate the feasibility of utilizing cornstalks to produce chaetoglobosin A by C. globosum W7 in solid-batch fermentation and to determine an optimal method for purification of the products. The output of chaetoglobosin A from the cornstalks was 0.34 mg/g, and its content in the crude extract was 4.80%. Purification conditions were optimized to increase the content of chaetoglobosin A in the crude extract, including the extract solvent, temperature, and pH value. The optimum process conditions were found to be acetone as the extractant, under room temperature, and at a pH value of 13. Under these conditions, a production process of the antifungal chaetoglobosin A was established, and the content reached 19.17%. Through further verification, cornstalks could replace crops for the production of chaetoglobosin A using this new production process. Moreover, the purified products showed great inhibition against Rhizoctonia solani, with chaetoglobosin A confirmed as the main effective constituent (IC50 = 3.88 µg/mL). Collectively, these results demonstrate the feasibility of using cornstalks to synthesize chaetoglobosin A and that the production process established in this study was effective.

Influence of Peanut Cultivars and Environmental Conditions on the Diversity and Community Composition of Pod Rot Soil Fungi in China

Peanut yield and quality are seriously affected by pod rot pathogens worldwide, especially in China in recent years. The goals of this study are to analyze the structure of fungal communities of peanut pod rot in soil in three peanut cultivars and the correlation of pod rot with environmental variables using 454 pyrosequencing. A total of 46,723 internal transcribed spacer high-quality sequences were obtained and grouped into 1,706 operational taxonomic units at the 97% similarity cut-off level. The coverage, rank abundance, and the Chao 1 and Shannon diversity indices of the operational taxonomic units were analyzed. Members of the phylum Ascomycota were dominant, such as Fusarium, Chaetomium, Alternaria, and Sordariomycetes, followed by Basidiomycota. The results of the heatmap and redundancy analysis revealed significant variation in the composition of the fungal community among the three cultivar samples. The environmental conditions in different peanut cultivars may also influence on the structure of the fungal community. The results of this study suggest that the causal agent of peanut pod rot may be more complex, and cultivars and environmental conditions are both important contributors to the community structure of peanut pod rot fungi.

Endophytic fungus strain 28 isolated from Houttuynia cordata possesses wide-spectrum antifungal activity

Abstract The aim of this paper is to identify and investigate an endophytic fungus (strain 28) that was isolated from Houttuynia cordata Thunb, a famous and widely-used Traditional Chinese Medicine. Based on morphological methods and a phylogenetic analysis of ITS sequences, this strain was identified as Chaetomium globosum. An antifungal activity bioassay demonstrated that the crude ethyl acetate (EtOAc) extracts of strain 28 had a wide antifungal spectrum and strong antimicrobial activity, particularly against Exserohilum turcicum (Pass.) Leonard et Suggs, Botrytis cinerea persoon and Botrytis cinerea Pers. ex Fr. Furthermore, the fermentation conditions, extraction method and the heat stability of antifungal substances from strain 28 were also studied. The results showed that optimal antifungal activity can be obtained with the following parameters: using potato dextrose broth (PDB) as the base culture medium, fermentation for 4–8 d (initial pH: 7.5), followed by extraction with EtOAc. The extract was stable at temperatures up to 80 °C. This is the first report on the isolation of endophytic C. globosum from H. cordata to identify potential alternative biocontrol agents that could provide new opportunities for practical applications involving H. cordata.

Chemical constituents from endophyte Chaetomium globosum in Imperata cylindrical / 中国中药杂志

Isolation and purification of chemical constituents from solid culture of endophyte Chaetomium globosum in Imperata cylindrical was performed through silica gel column chromatography, gel filtration over Sephadex LH-20 and preparative HPLC. Nine compounds were obtained and their structures were determined as chaetoglobosin F(1), chaetoglobosin Fex(2), chaetoglobosin E(3) cytoglobosin A(4), penochalasin C(S), isochaetoglobosin D (6), N-benzoylphenylalaninyl-N-benzoyphenylalaninate(7), uracil(8) and 5-methyluracil(9), respectively, based on HR-MS and NMR data and comparison with literatures. Compound 7 was isolated from Chaeeomium sp. for the first time. In vitro cytotoxicity of compounds was evaluated using MTT mothed and 1,3,4 and 5 showed inhibition activity to the human cervical carcinoma cell HeLa with IC50 values of 99.43, 23.77, 97.92, 86.25 micromol x L(-1), while positive cotolocisnin Ad apno1ch alse IC50 24.33 micromol x L(-1).

Biological Control of Phytophthora palmivora Causing Root Rot of Pomelo Using Chaetomium spp

Phytophthora diseases have become a major impediment in the citrus production in Thailand. In this study, an isolate of Phytophthora denominated as PHY02 was proven to be causal pathogen of root rot of Pomelo (Citrus maxima) in Thailand. The isolate PHY02 was morphologically characterized and identified as Phytophthora palmivora based on molecular analysis of an internal transcribed spacer rDNA sequence. This work also presents in vitro evaluations of the capacities of Chaetomium spp. to control the P. palmivora PHY02. As antagonists, Chaetomium globosum CG05, Chaetomium cupreum CC3003, Chaetomium lucknowense CL01 inhibited 50~61% mycelial growth, degraded mycelia and reduced 92~99% sporangial production of P. palmivora PHY02 in bi-culture test after 30 days. Fungal metabolites from Chaetomium spp. were tested against PHY02. Results showed that, methanol extract of C. globosum CG05 expressed strongest inhibitory effects on mycelial growth and sporangium formation of P. palmivora PHY02 with effective dose ED50 values of 26.5 microg/mL and 2.3 microg/mL, respectively. It is interesting that C. lucknowense is reported for the first time as an effective antagonist against a species of Phytophthora.

Efficacy of Chaetomium Species as Biological Control Agents against Phytophthora nicotianae Root Rot in Citrus

Thailand is one of the largest citrus producers in Southeast Asia. Pathogenic infection by Phytophthora, however, has become one of major impediments to production. This study identified a pathogenic oomycete isolated from rotted roots of pomelo (Citrus maxima) in Thailand as Phytophthora nicotianae by the internal transcribed spacer ribosomal DNA sequence analysis. Then, we examined the in vitro and in vivo effects of Chaetomium globosum, Chaetomium lucknowense, Chaetomium cupreum and their crude extracts as biological control agents in controlling this P. nicotianae strain. Represent as antagonists in biculture test, the tested Chaetomium species inhibited mycelial growth by 50~56% and parasitized the hyphae, resulting in degradation of P. nicotianae mycelia after 30 days. The crude extracts of these Chaetomium species exhibited antifungal activities against mycelial growth of P. nicotianae, with effective doses of 2.6~101.4 microg/mL. Under greenhouse conditions, application of spores and methanol extracts of these Chaetomium species to pomelo seedlings inoculated with P. nicotianae reduced root rot by 66~71% and increased plant weight by 72~85% compared to that in the control. The method of application of antagonistic spores to control the disease was simple and economical, and it may thus be applicable for large-scale, highly effective biological control of this pathogen.

Isolamento de Chaetomium spp. em lesões subcutâneas de um cão: relato de caso / Isolation of Chaetomium spp. on subcutaneous lesions of a dog: case report

Caracterizou-se clinicamente a infecção pelo Chaetomium spp. em um cão, e descreveu-se seu isolamento e identificação. Ao exame dermatológico foram observadas pápulas nas orelhas, no tronco lateral e nos membros pélvicos. Ao rompimento de uma dessas pápulas, fluiu um líquido serosanguinolento com consequente úlcera no local. Foi colhido material para isolamento micológico, por meio de raspado das pápulas da orelha e da cauda. O diagnóstico foi micose subcutânea por Chaetomium spp.

This work aimed to clinically characterize the infection by Chaetomium spp. in a dog, as well as describe its isolation and identification. Upon dermatological exam, papules on ears, lateral trunk and pelvic members were noticed. After the disruption of these papules there was serosanguineous secretion flowed by consequent ulcer in the region. Material for mycological isolation was picked, and a scraping of papules from ear and tail was done. The diagnosis was subcutaneous mycosis caused by Chaetomium spp.

possible therapeutic effect of 'chaetomium globosum' fungal extract on experimentally induced rheumatoid arthritis

Rheumatoid arthritis [RA] is an autoimmune disease that leads to chronic inflammation in the joints with subsequent cartilage and bone destruction. RA induces a massive burden on health services worldwide. This study was designed to evaluate the possible therapeutic effect of 'Chaetomium globosum' extract in the treatment of RA in a rat model. Forty male Wistar albino rats aged 8-10 weeks were divided into four groups of 10 rats each. The control group [group I] was injected subcutaneously with 0.1 ml of saline. RA was induced in the other three groups [groups II, III, and IV] by single subcutaneous injection of 0.1 ml of complete Freund's adjuvant in the footpad of the right hind paw. Group II was induced with arthritis and left untreated. Rats in groups III and IV were treated by administering either C. globosum 10 micro g/kg or methotrexate [MTX] 0.3 mg/kg subcutaneously twice weekly for 2 weeks from day 12 after induction of arthritis. Animals of all groups were sacrificed on day 28 from the start of the experiment. The ankle joints were processed and stained with H and E, Masson's trichrome, and immunohistochemical stain for inducible nitric oxide synthase. Specimens were also processed for transmission electron microscopic study. Untreated arthritic rats revealed paw swelling, synovial hyperplasia, inflammatory infiltration, collagen accumulation, cartilage degradation, bone destruction, and significant increase in inducible nitric oxide synthase-positive chondrocytes. Transmission electron microscopic examination confirmed these results. The present study demonstrated for the first time that C. globosum significantly reduced all the clinical and histopathological changes of arthritis similar to MTX. C. globosum extract had therapeutic effects similar to the well-established drug MTX and could be devoid of its serious side effects.

Isolation, idetification and anti-HIV-1 integrase activity of culturable endophytic fungi from Tibetan medicinal plant Phlomis younghusbandii Mukerjee / 药学学报

A total of 52 endophytic fungi were isolated from roots and stems of Tibetan medicinal plant Phlomis younghusbandii Mukerjee. These fungal isolates were molecularly identified based on ITS sequnces and 28S sequences distributed to 12 genera, including Phoma, Chaetosphaeronema, Fusarium and Leptosphaeria, etc. Among them, the dominant genus was Phoma. Extracts of all strains were evaluated for anti-HIV-1 integrase activity by using soluable integrase expressed in E. coli BL21 (DE3). The results showed that seven samples from five fungal endophytes PHY-24, PHY-38, PHY-40, PHY-51, PHY-53, which belonged to genus Chaetosphaeronema, inhibited strand transfer reaction catalyzed by HIV-1 integrase with IC50 values, of 6.60, 5.20, 2.86, 7.86, 4.47, 4.56 and 3.23 microg x mL(-1) respectively. In conclusion, the endophytic fungi of Phlomis younghusbandii Mukerjee are valuable for further screening anti-HIV-1 integrase agents.

Onychomycosis Caused by Chaetomium globosum

Onychomycosis is usually caused by dermatophytes, but some nondermatophytic molds and yeasts are also associated with invasion of nails. The genus Chaetomium is a dematiaceous nondermatophytic mold found in soil and plant debris as a saprophytic fungus. We report the first Korean case of onychomycosis caused by Chaetomium globosum in a 35-year-old male. The patient showed brownish-yellow discoloration and subungual hyperkeratosis on the right toenails (1st and 5th) and left toenails (1st and 4th). Direct microscopic examination of scraping on the potassium hydroxide preparation revealed septate hyphae and repeated cultures on Sabouraud's dextrose agar (SDA) without cycloheximide slants showed the same fast-growing colonies, which were initially velvety white then turned to dark gray to brown. However, there was no growth of colony on SDA with cycloheximide slants. Brown-colored septated hyphae, perithecia and ascospores were shown in the slide culture. The DNA sequence of internal transcribed spacer region of the clinical sample was a 100% match to that of C. globosum strain ATCC 6205 (GenBank accession number EF524036.1). We confirmed C. globosum by KOH mount, colony, and light microscopic morphology and DNA sequence analysis. The patient was treated with 250 mg oral terbinafine daily and topical amorolfine 5% nail lacquer for 3 months.

Búsqueda y selección de una proteasa fúngica con potencial aplicación en la restauración de documentos históricos en el Archivo de Bogotá / Screening for a fungal protease with potential in the biorestoration of historical valuable documents in Bogota Archive

Se ha buscado y seleccionado sistemáticamente una proteasa que pudiese ser usada en la eliminación "limpia" de encolantes sobre soportes documentales con valor de patrimonio histórico de forma eficiente y económica, a partir de la colección de hongos filamentosos del Archivo de Bogotá. De 74 morfotipos viables evaluados sobre placas selectivas, 32 morfotipos presentaron formación de halos de hidrólisis evidentes sobre placas diferenciales. De ellos, se evaluó el perfil isoenzimático de 8 morfotipos provenientes de muestreos documentales directos y de 2 morfotipos proteolíticos promisorios provenientes de un trabajo previo. Los 10 morfotipos seleccionados fueron representativos de los géneros Penicillium, Stachybotrys, Chaetomium, y Eladia. Luego de inducir la producción de proteasas extracelulares en medios líquidos diferenciales bajo tres fases de fermentación, se realizaron isoelectroénfoques analíticos tendientes a la observación de isoformas en el gradiente de pH establecido (3.0-10.0). Solo los morfotipos 8D (Chaetomium sp.) y 21D (Eladia saccula) presentaron una isoforma alcalina extrema, de puntos isoeléctricos 8.5 y 8.8, respectivamente, susceptible de selección con miras a su purificación y caracterización parcial de forma económica y eficiente. Los demás morfotipos, representativos de los géneros Penicillium sp., y Stachybotrys sp., presentaron unicamente isoformas proteolíticas en el rango acido de pH con puntos isoeléctricos que oscilan entre 4.0 y 5.0.

Studies on a protease as an efficient, environmental friendly and relatively economical remover of residual proteins for historical valuable documents were performed and was selected for this work, from the filamentous fungi collection of the Bogota Archive. 32 morphotypes of 74 evaluated show hydrolytic activities over differential solid media. From them, 8 morphotypes obtained directly from documental samples and representative of the genera Penicillium and Stachybotrys were selected and their isoenzyme profile were tested. Also 2 previous morphotypes with promisorius proteolytic activities and representative of the genera Chaetomium and Eladia were analysed. Extracelullar proteases production was induced in differential liquid media on three fermentation steps and analitycal isoelectrofocusing were performed over pH 3.0-10.0 ranges. Only the morphotypes 8D (Chaetomium sp.), and 21D (Eladia sp.), showed an alkaline isoform with pIs 8.5 and 8.8, respectly, suceptible of selection for its purification and characterization through efficient and economical way. The others morphotypes only showed acid isoforms with pIs in the range of 4.0 and 5.0.

Comparative study for the kinetics of extracellular xylanases from Trichoderma harzianum and chaetomium thermophilum

Xylanases assume special importance in the paper and pulp industry as they replace toxic chemicals such as chlorine and chlorine dioxide for developing eco-friendly processes. This study evaluated xylanases produced by two fungi, the mesophilic fungus Trichoderma harzianum and a thermophilic fungus Chaetomium thermophilum. Among the polymeric substrates studied for xylanase production by both the fungi, birch wood xylan was found to be the best inducer of xylanases. Xylanases induction was subject to glucose repression. Partially purified xylanases preparation from T. harzianum and C. thermophilum exhibited optimal activities at pH 5 and 6 and at 60ºC and 70ºC, respectively. The apparent Km and Vmax values for the partially purified xylanase from T. harzianum using oat spelt xylan as a substrate were 4.8 mg mL-1 and 0.526 µmol min-1 mg-1, respectively. Whereas values of the partially purified xylanase from C. thermophilum were 2.96 mg mL-1 and 0.25 µmol min-1 mg-1, respectively. These findings in this study have great implications for the future applications of xylanases.

Onychomycosis Due to Nondermatophytic Molds

BACKGROUND: Although there have been many studies about onychomycosis due to nondermatophytic molds (NDM), few studies about etiologic agents including NDM in onychomycosis have been reported in Korea. Objective: This study investigated onychomycosis due to NDM in the Gyeongju area of Korea. OBJECTIVE: This study investigated onychomycosis due to NDM in the Gyeongju area of Korea. METHODS: In the 10-year period from 1999~2009, we reviewed 59 patients with onychomycosis due to NDM. The etiologic agents were identified by cultures on Sabouraud's Dextrose agar with and without cycloheximide. In some cases, internal transcribed spacer sequence analysis was done. NDM isolated considered pathogens when the presence of fungal elements was identified by direct microscopy observation and in follow-up cultures yielding the same fungi. RESULTS: Onychomycosis due to NDM comprised 2.3% of all onychomycosis. Of the 59 patients with onychomycosis due to NDM, 84.7% were toenail onychomycosis and 15.3% were fingernail onychomycosis. The incidence rate was highest in the fifth decade (27.1%). The ratio of male to female patients was 1:1.6. The frequency of associated diseases, in descending order, was hypertension, diabetes mellitus, and cerebral hematoma. Distal and lateral subungual onychomycosis (86.4%) was the most common clinical type of onychomycosis. Aspergillus spp. was the most frequently isolated etiologic agent of onychomycosis due to NDM (83.0%). Other causative agents were Scopulariopsis brevicaulis (10.2%), Acremonium spp. (3.4%), Fusarium solani (1.7%), and Chaetomium globosum (1.7%). CONCLUSION: Because of the increase in onychomycosis due to NDM, we suggest the need of a careful mycological examination in patients with onychomycosis.

Effect of PEG stress on plantlets of Chrysanthemum morifolium induced by endophytic botrytis sp. (C1) and Chaetomium globosum (C4) / 中国中药杂志

The effect of the endophytic fungi Botrytis sp. (C1) or Chaetomium globosum (C4) on the drought resistance of Chrysanthemum morifolium was studied. Ch. morifolium plantlets were inoculated with C1, C4 and cultured in the pots for 60 days, then the plantlets were stressed by 0%, 10%, 20%, 30%, 40% PEG6000 respectively in order to simulate different drought conditions. Biomass, the activities of SOD, POD, PAL, the contents of MDA and soluble protein of each group were determined. The results showed that endophytic fungi groups grew better than the control (without inoculation endophytic fungi). With the increasing of the concentration of PEG6000, the biomass of Ch. morifolium of each groups decreased, while the biomass of fungi groups was significantly higher than that of control, moreover C4 group higher than C1 group. With the concentration of PEG increasing, the content of MDA of each group increased too, while POD activity and soluble protein content of all treatments increased at first and then decreased. SOD activity and PAL activity of the control were increased with the increase of PEG concentration, but SOD activity of the two fungi groups were stable. After been stressed by different concentrations of PEG, MDA content of two fungi groups were always lower than the control, while SOD activity, POD activity, PAL activity and soluble protein content were higher. In conclusion, endophytic fungi can increase the drought resistance of Ch. morifolium.

Molecular characterization of Chaetomium species using URP-PCR

Chaetomium spp. are common colonizers of soil and cellulose-containing substrates. Seventeen isolates of Chaetomium spp., which included 15 isolates of C. globosum and one each of C. reflexum and C. perlucidum, were genetically characterized with universal rice primers (URP - primers derived from DNA repeat sequences in the rice genome) using polymerase chain reaction (URP-PCR). Out of the 12 URP's used in the study, nine primers were effective in producing polymorphic fingerprint patterns from DNA of Chaetomium spp. Analysis of the entire fingerprint profile using the unweighted pair-group method with arithmetic averages (UPGMA) clearly differentiated C. globosum isolates from C. perlucidum and C. reflexum. One of the primers, URP-2R, produced a uniform DNA band of 1.9 kb in all the isolates of C. globosum but not in C. perlucidum and C. reflexum, which can be used as molecular marker to differentiate C. globosum from other species. Our results indicate that URP's are sensitive and give reproducible results for assaying the genetic variability in Chaetomium spp.

Cloning and expressing of cellulase gene (cbh2) from thermophilic fungi Chaetomium thermophilum CT2 / 生物工程学报

Chaetomium thermophilum CT2 can produce extracellular cellulase with industrial value. We designed two degenerate primers to amplify catalytic domain sequence of cellobiohydrolase II ( CBH II). Full length of cDNA was obtained by rapid amplification of cDNA ends technologies. DNA sequencing revealed that cbh2 has an open reading frame of 1428bp, which encodes a putative polypeptide of 476 amino acids. The deduced amino acid sequence shows that the predicted molecular mass is 53 kD and the cbh2 consists of a fungal-type carbohydrate binding domain (CBD) separated from a catalytic domain by a linker region rich in proline/serine/threonine. PCR product consisting of the entire CBH II coding region without its signal sequences was cloned into the yeast secretive plasmid pPIC9K, which was then transformed into Pichia pastoris GS115. Highly efficient production of the cellobiohydrolase II was achieved in P. pastoris under the control of the AOX1 promoter, and the expressing level was 1.2 mg/mL by small-scale culturing. The recombinant cellobiohydrolase II was purified by using ammonium sulfate fraction, DEAE-Sepharose Fast flow chromatography. A molecular mass of the purified enzyme is 67 kD determined by SDS-PAGE and this is similar to the native cellobiohydrolase II purified from C. thermophilum CT2. The recombinant enzyme exhibited optimum catalytic activity at pH 4.0 and 50 degrees C respectively. It was thermostable at 50 degrees C and retained 50% of its original activity after 30 min at 70 d degrees C . The high level of fully active recombinant cellobiohydrolase II got from P. pastoris makes this expression system attractive for fermentor and industrial applications.

Distribution of Fungi in the Sandy Soil of Egyptian Beaches

The mycobiota of the sandy soil of Egyptian beaches was investigated in thirty six sand samples collected from nine different localities in Egypt. The filamentous fungi were identified and assigned to thirty one genera and fifty one species. Greater populations as well as a wider spectrum range of fungal genera and species were obtained in sandy soil of Alexandria beach while Balteem beach was the poorest one. The total count of the genus or species was not depended upon cases of isolation. Most of the genera detected belonged to the Deuteromycotina with fewer proportions belonging to the Ascomycotina and Zygomycotina. The genera of highest incidence and their respective numbers of species were: Penicillium (35.72%, 6 species) and Aspergillus (30.28%, 16 species). The species which showed the highest incidence in all cases was P. chrysogenum, followed by P. citrinum, A. flavus, Chaetomium murorum and Trichoderma viride. A few number of other genera and species were also detected.