Efficiency of developed solid state bioreactor ‘FERMSOSTAT’ on cellulolytic and xylanase enzymes production

FERMSOSTAT is a developed laboratory scale solid state fermenter. It is a horizontal stirrer drum bioreactor with about 70 L capacities. The fermenter is made of stainless steel which is anti-corrosive and non-toxic to the process organism. The fermenter is equipped with sets of control systems for temperature, agitation, aeration and also outlets for substrate sampling as well as inlets for inoculation and substrate additions. The uniqueness of this FERMSOSTAT system is its ability to carry out in situ substrate sterilization and extraction of enzymes at the end of SSF process. Moreover, the mixing system provided by FERMSOSTAT can be performed either full or half mixing as well as forward or reverse mixing. Furthermore, the mixing can be programmed to run at certain agitation rate and time interval during the fermentation process to prevent or reduce damage to the fungus mycelia. FERMSOSTAT is a developed SSF bioreactor and not an improvement of any existing one. The performances of FERMSOSTAT have been evaluated. Under optimum solid state fermentation conditions, about 63.4, 397 and 3.21 U/g of CMCase, xylanase and FPase activities were detected, which were higher compared to the tray system

ANTI-MRSA OF THE ETHYL ACETATE CRUDE EXTRACT FROM LASIODIPLODIA PSEUDOTHEOBROMAE IBRL OS-64, AN ENDOPHYTIC FUNGUS ISOLATED FROM LEAF OF OCIMUM SANCTUM LINN

Objective: To investigate the effects of ethyl acetate crude extract of an endophytic fungus, L. pseudotheobromae IBRL OS-64 which was isolated from leaf of Ocimum sanctum against the growth of methicillin-resistant Staphylococcus aureus (MRSA)., a common pathogenic bacteria to human.Methods: Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined. Time-kill assay was used to examine the effect of the extract on the MRSA growth profile. The effects of the extract on the ultrastructure of MRSA cells were analyzed by scanning electron microscopic (SEM) study.Results: The results obtained from this study showed the fungal ethyl acetate crude extract exhibited a strong bactericidal effect on MRSA where the ratio of MBC/MIC was 2 and less than 4. The MIC and MBC values were 125.0 and 250.0 µg/ml, respectively. The time-kill study revealed that the bacteriocidal activity of the extract was both concentration and time-dependent. After 12 h treatment, the interaction of extract with MRSA cells resulted in the formation of dents, cavity or small dimples on the cell surface, indicating disintegration of the cell wall and cell membrane that resulting in leakage of their cytoplasmic contents, and ultimately cell death.Conclusion: The ethyl acetate crude extract of L. pseudotheobromae IBRL OS-64 showed a significant anti-MRSA activity and principally affected the cell wall and the cell membrane of the growing MRSA cells. This is the first report on L. pseudotheobromae, an endophytic fungus isolated from medicinal herb, Ocimum sanctum Linn

Assessment of double screening programmes via solid substrate fermentation (SSF) in a flask system and identification of lovastatin potential producer

Local economical substrates namely rice bran and unprocessed brown rice was applied into fermentation condition to produce a potent secondary metabolite compound, lovastatin. A basis condition of fermentation viz. 70% (v/w) of moisture content (adjusted to pH 6.0), 1x107 spore/ml of inoculum size, mixture of 1:1 substrates and 7 days of incubation period, was applied into SSF system. During a preliminary test, all of 72 fungi disclosed positive dark spot onto the thin layer chromatography plate (TLC). In order to verify the existence of lovastatin, the secondary screening which involving high performance liquid chromatography (HPLC) was conducted. Out of 72, only 71 fungi were detected as lovastatin producers and the highest production was stated from SAR I isolate with 68.72±0.84 mg lovastatin/g dry substrate and 0.87±0.03 mg glucosamine/g dry substrate of fungal growth. SAR I isolate was identified via colony and microscopic morphologies. Through the observations, SAR I isolate was identical to Aspergillus nige

The Influence of Physical Parameters Towards Hyper Cholesterol Reducing Agent Production, Lovastatin, Under Solid Substrate Fermentation (SSF) Condition

Two potential substrates namely rice bran and unprocessed brown rice indicated positive result of lovastatin existence. Aspergillus Niger SAR I, our local isolated fungus, took a responsibility to cooperate with those substrates in SSF system. Further experiment including initial profile production, effect of physical parameters (temperature, inoculum size and substrate quantity) and final profile production, were carried out. For initial profile, a basic condition of SSF which consisted 70% (v/w) of moisture content (adjusted to pH 6.0), 5 g substrates mixture (ungrounded size), 1x107 spore/ml of inoculum size and incubation temperature at 30±2 0C, was conducted in a flask system and fermented for 7 days. Those conditions allowed 160.03±3.79 mg lovastatin/g dry substrate of lovastatin production during initial stage. After a study of effect of physical parameters, it showed that the optimum temperature was still at ambient temperature (30±2 0C) and substrate quantity of 5 g but different inoculum size (1x105 spore/ml). Each parameters specifically temperature, inoculum size and substrate quantity produced 253.98±5.92 mg lovastatin/g dry substrate, 297.64±0.56 mg lovastatin/g dry substrate and 298.72±44.12 mg lovastatin/g dry substrate, respectively. Throughout the final profile, the production was 305.08±14.15 mg lovastatin/g dry substrate which made the total increment hit to almost 91%. In this experiment, lovastatin was subjected into high performance liquid chromatography (HPLC) with acetonitrile and phosphoric acid (pH 3.0) as a mobile phase

Involvement of Physical Parameters in Medium Improvement for Tannase Production by Aspergillus niger FETL FT3 in Submerged Fermentation

Aspergillus niger FETL FT3, a local extracellular tannase producer strain that was isolated from one of dumping sites of tannin-rich barks of Rhizophora apiculata in Perak, Malaysia. This fungus was cultivated in 250 mL Erlenmeyer flask under submerged fermentation system. Various physical parameters were studied in order to maximize the tannase production. Maximal yield of tannase production, that is, 2.81 U per mL was obtained on the fourth day of cultivation when the submerged fermentation was carried out using liquid Czapek-Dox medium containing (percent; weight per volume) 0.25% NaNO3, 0.1% KH2PO4, 0.05% MgSO4 ·7H2O, 0.05% KCl, and 1.0% tannic acid. The physical parameters used initial medium pH of 6.0, incubation temperature of 30°C, agitation speed of 200 rpm and inoculums size of 6 × 106 spores/ ml. This research has showed that physical parameters were influenced the tannase production by the fungus with 156.4 percent increment

PU.1 opposes IL-7-dependent proliferation of developing b cells with involvement of the direct target gene bruton tyrosine kinase

Deletion of genes encoding the E26 transformation-specific transcription factors PU.1 and Spi-B in B cells (CD19-CreΔPB mice) leads to impaired B cell development, followed by B cell acute lymphoblastic leukemia at 100% incidence and with a median survival of 21 wk. However, little is known about the target genes that explain leukemogenesis in these mice. In this study we found that immature B cells were altered in frequency in the bone marrow of preleukemic CD19-CreΔPB mice. Enriched pro-B cells from CD19-CreDPB mice induced disease upon transplantation, suggesting that these were leukemia-initiating cells. Bone marrow cells from preleukemic CD19-CreΔPB mice had increased responsiveness to IL-7 and could proliferate indefinitely in response to this cytokine. Bruton tyrosine kinase (BTK), a negative regulator of IL-7 signaling, was reduced in preleukemic and leukemic CD19-CreΔPB cells compared with controls. Induction of PU.1 expression in cultured CD19-CreΔPB pro-B cell lines induced Btk expression, followed by reduced STAT5 phosphorylation and early apoptosis. PU.1 and Spi-B regulated Btk directly as shown by chromatin immunoprecipitation analysis. Ectopic expression of BTK was sufficient to induce apoptosis in cultured pro-B cells. In summary, these results suggest that PU.1 and Spi-B activate Btk to oppose IL-7 responsiveness in developing B cells

PU.1 opposes IL-7-dependent proliferation of developing b cells with involvement of the direct target gene bruton tyrosine kinase

Deletion of genes encoding the E26 transformation-specific transcription factors PU.1 and Spi-B in B cells (CD19-CreΔPB mice) leads to impaired B cell development, followed by B cell acute lymphoblastic leukemia at 100% incidence and with a median survival of 21 wk. However, little is known about the target genes that explain leukemogenesis in these mice. In this study we found that immature B cells were altered in frequency in the bone marrow of preleukemic CD19-CreΔPB mice. Enriched pro-B cells from CD19-CreDPB mice induced disease upon transplantation, suggesting that these were leukemia-initiating cells. Bone marrow cells from preleukemic CD19-CreΔPB mice had increased responsiveness to IL-7 and could proliferate indefinitely in response to this cytokine. Bruton tyrosine kinase (BTK), a negative regulator of IL-7 signaling, was reduced in preleukemic and leukemic CD19-CreΔPB cells compared with controls. Induction of PU.1 expression in cultured CD19-CreΔPB pro-B cell lines induced Btk expression, followed by reduced STAT5 phosphorylation and early apoptosis. PU.1 and Spi-B regulated Btk directly as shown by chromatin immunoprecipitation analysis. Ectopic expression of BTK was sufficient to induce apoptosis in cultured pro-B cells. In summary, these results suggest that PU.1 and Spi-B activate Btk to oppose IL-7 responsiveness in developing B cells

Effects of Cultural Conditions in Enhancing the Production of Anti-MRSA Activity of Lasiodiplodia pseudotheobromae IBRL OS-64, an Endophytic Fungus Isolated from Leaf of Ocimum sanctum L. in Submerged Fermentation System

The effects of cultural conditions on the production of anti-MRSA activity of the endophytic fungal isolate, Lasiodiplodia pseudotheobromae IBRL OS-64 isolated from the leaf of a local medicinal herb, Ocimum sanctum Linn. was studied. The highest anti-MRSA activity of 50.0±0.1 U/mL and 1.46±0.1 g/L of fungal growth were obtained after incorporating all the improved cultural conditions which consisted of yeast extract sucrose broth supplemented with host plant extract, with initial medium pH of 6, 2 mycelial plugs of 4 days old seed culture, cultivation temperature of 30°C and cultivated for 16 days under static conditions without the presence of light. After improvement of cultural conditions, a significant increment of anti-MRSA activity of about 57.57% (2.4 folds) and a slight increment of fungal growth of about 8.15% (1.1 folds) were obtained compared to the cultural condition before improvement. Indeed the improvement of cultural conditions greatly affected the anti-MRSA activity and growth production by L. pseudotheobromae IBRL OS-64 isolate