Eupenicillium Javanicum Research Articles

Evaluation of feed supplementation using legume leaf meal and spent substrate fermented by <i>Eupenicillium javanicum</i> as a potential feed for ruminants: an <i>in vitro</i> study

Evaluation of feed supplementation using legume leaf meal and spent substrate fermented by <i>Eupenicillium javanicum</i> as a potential feed for ruminants: an <i>in vitro</i> study

Endophytic fungi from Dongxiang wild rice (Oryza rufipogon Griff.) show diverse catalytic potential for converting glycyrrhizin.

Endophytic fungi inhabiting niche environments are novel biocatalyst resources that need to be exploited urgently. In this study, 63 endophytic fungi isolated from Dongxiang wild rice (Oryza rufipogon Griff.) were tested to assess their potentials to transform glycyrrhizin (GL) into glycyrrhetinic acid monoglucuronide (GAMG) or glycyrrhetinic acid (GA), of which 12 strains were shown to have β-d-glucuronidase activity. Based on morphological characteristics and rDNA ITS sequence analysis, the strains S59, L138, L55 and R57 with high GL molar conversion rates (55%, 45%, 65% and 89%) were further identified as Microsphaeropsis arundinis S59, Penicillium rubens L138, Aspergillus flavus L55 and Eupenicillium javanicum R57, respectively. These four strains with four different types of GL conversion processes were identified, i.e., (1) GL → GAMG in M. arundinis S59, (2) GL → GAMG and GA in A. flavus L55, (3) GL → GA in P. rubens L138, and (4) GL → GAMG → GA in E. javanicum R57, in which the bioconversion type (4) is reported for the first time. The study not only provided abundant and diverse β-d-glucuronidase resources that can be used for GL bioconversion, especially for GAMG biosynthesis from endophytic fungi, but also expanded our knowledge of potential roles of endophytes as new biocatalysts in biotransformation.

Fungal metalloprotease generate whey-derived peptides that may be involved in apoptosis in B16F10 melanoma cells

Proteases are enzymes that act in the hydrolysis of proteins and have several industrial applications. Moreover, proteases have gained prominence as enzymes for the generation of bioactive peptides from the hydrolysis of different protein sources. Milk is the most studied protein source to obtain peptides due to its nutritional and physiological effects and has been studied as complementary therapeutic approaches for the cancer treatment, interacting specifically with cancer cells, consequently fewer side effects. The ability of Eupenicillium javanicum metalloprotease to generate whey-derived peptides with antioxidant activity has already been demonstrated. For this reason, we thus hypothesized that whey-derived peptides from Eupenicillium javanicum metalloprotease hydrolysis could also have a potential against melanoma cell lines. In this study, B16F10 melanoma cells were treated for 72 h with whey-derived peptides and the effects on cell viability were determined. Moreover, the protein profiles of the treated and nontreated cells were compared in proteomic assay and mass spectrometry analyzes. Whey-derived peptides impaired about 62% cell viability, and proteomic approach associated this behavior to modulate proteins involved in proliferation, energy, apoptosis, metastatic and malignancy rates. This study describes the relevance of microbial enzymes in generation of whey-derived peptides with biological activity against melanoma cells.

Resistance of Eupenicillium javanicum mold spores to the light-emitting diode (LED), LED-assisted thermal and thermal processing in strawberry and apple juices

The use of light-emitting diode (LED) technology for the non-thermal processing of foods is a major topic of interest among various research groups. This study is aimed at inactivating the Eupenicillium javanicum ascospores present in strawberry and apple juices using a combination of a visible LED (vis-LED, 430–630 nm, 216–420 J/cm2) and 90 °C thermal treatment, as well as to compare the findings with the inactivation done using thermal-processes alone. The results showed that violet-blue LEDs within the range of 430 and 460 nm with an energy between 300 and 420 J/cm2 were better for the inactivation of E. javanicum ascospores than the green and red LEDs which were within the 550–630 nm region with an energy range from 216 to 264 J/cm2. Furthermore, the inactivation process conducted using vis-LED was affected by the juice's soluble solid contents and the calculated DLED-values were within the range of 116.3 J/cm2 to 277.8 J/cm2 in juices with a Brix scale value of 10–20°. Finally, the inactivation rate obtained from combining a violet-blue LED with a 90 °C thermal treatment was similar to the rate of using the thermal treatment alone.

Use of Pulsed Electric Field for the Inactivation of Eupenicillium Javanicum Ascospores in Pineapple Juice

Heat resistant molds are principle spoilage agents in foods and beverages with low acidity. The main objective of this work was to investigate the effect of 65 kV/cm PEF on the log reductions of Eupenicillium javanicum ascospores in 10–30°Brix pineapple juice as well as the modelling. Then, the first-order and Weibull parameters of the 65 kV/cm PEF inactivation of E. javanicum ascospores was estimated and compared. Further, the effect of PEF in combination with ultraviolet (UV) light treatment on the log reductions of E. javanicum ascospores was studied. Decreasing the soluble solid content of the juice from 30 to 10°Brix for 11.3 pulses increased the spore inactivation in pineapple juice by 2.7 log. A pulse number of 16 would be required by the 65 kV/cm PEF to achieve a 5-log reduction in juice. The Weibull model described spore inactivation by pulsed electric field. The estimated b-values for the 65 kV/cm PEF were 0.673 at 10°Brix, 0.041 at 20°Brix and 0.010 at 30°Brix, with n-values between 0.73 and 2.08. Although the combination of the PEF and UV light resulted in a slightly greater microbial inactivation, however two hurdles were not suggested. The results of this study confirmed the advantage of PEF technology for the inactivation of E. javanicum ascospores in pineapple juice.

The stability of the BS4 enzymes activity on the process of feed pelleting

BS4 mannanase enzyme can increase the digestibility of chicken feed containing 20% palm kernel cake. The palm kernel cake was produced by Eupenicillium javanicum molds through coconut cake solid substrate fermentation. In its application as a feed additive, the stability of activity is something that must be considered. This study was conducted to evaluate the stability of the saccharification activity of BS4 enzymes against heating around 90°C in the feed pelleting process. The treatment was used with and without pelleting the process of starter and finisher phases broiler feed with BS4 and commercial enzymes’ addition. The parameter measured was the activity of saccharification of palm kernel cake. The results showed that the pelleting process is very influential on BS4 enzymes and can reduce 65.8 to 68.4% of its saccharification activity. While on commercial enzymes, the decrease is about 6%. It can be concluded that the BS4 enzyme is very susceptible to heat in the pelleting process so that technology is needed to protect the stability of its activity.

Production of Cellulase and Xylanase from Eupenicillium Javanicum by Solid-State Fermentation Utilizing Pineapple Crown Leaves Waste as the Substrate

Vast applications of cellulases and xylanases in many fields over the last few decades and high production cost demand extensive research in improving its quality and finding low cost substrates. These includes pulp and paper, food and feed, brewing, agriculture and biofuel industries. The main objective of this study was to investigate the use of pineapple crown leaves waste as the substrate to produce cellulases and xylanases in solid-state cultures (SSF) of Eupenicillium javanicum InaCC F154. Three SSF temperatures (25°C, 30°C and 35°C) and pH (4.0, 6.0 and 8.0) were tested for 96 h in solid media containing defatted soybean, (NH4)2SO4, NaNO3, CaCl2, and small amount of H2O. Then, its comparison with submerged fermentation (SmF) was also carried out. The SSF results showed that increasing temperature from 25°C to 35°C, increased the production of these enzymes. The pH also affected its activities, except at pH 4.0. The optimum pH and temperature for cellulase and xylanase production were found to be 6.0 and 35°C, respectively. The maximum cellulase obtained was 0.261 U/mL while much higher was obtained for xylanase with activity of 1.683 U/mL. SSF was better than SmF for xylanases (0.501 U/mL vs. 1.683 U/mL). The outcome of this study exhibits the potential of pineapple crown leaves as the substrate for xylanase production from E. javanicum by solid-state fermentation.

Thermal Inactivation of Eupenicillium Javanicum Ascospores in Pineapple Juice: Effect of Temperature, Soluble Solids and Spore Age

Food spoilage leading to food waste and substantial economic losses is a major issue for the food industry. The spoilage is often due to bacterial and fungal contamination in foods. Spores from microorganism pose a great concern since they can survive pasteurization and grow in specific foods. The obj1ectives of this work were to investigate the effect of temperature (T: 80-90°C), soluble solid concentration (11–20°Brix), and spore age (30–60 days) on the log reductions of Eupenicillium javanicum ascospores in pineapple juice. It was also carried out to estimate the first-order kinetic parameters (D and z-values) from the log survivor curves. Increasing the temperature from 80 to 90°C for 10 min increased the spore inactivation in 11°Brix pineapple juice by 6.4 log. For 5-log inactivation, increasing the percentage weight (% wt.) of sucrose from 11°Brix to 20°Brix at 90°C, increased the time needed to inactivate the spores by 2.6 min. Likewise, increasing the age of ascopores from 30 days to 60 days, also increased the time required by 16.7 min. The estimated D-values for 30-days old spores in 11°Brix juices were 19.84 min at 80°C, 4.99 min at 85°C and 1.45 min at 90°C, with z-values of 8.6-8.8°C. The results obtained provide useful data to design and predict pasteurization process targeting E. javanicum ascospores.

Optimization of BS4 Enzyme production with Different Substrate Thickness and Type of Trays

BS4 enzyme that is produced from solid substrate fermentation (SSF) on coconut cake with Eupenicillium javanicum BS4 in tray bioreactor has been applied as a feed additive. It increases the nutritional value of animal feedstuff. The BS4 production on SSF may be influenced by the better aeration through the perforated trays or by the thinner substrate. The aim of this research is to optimize the production of BS4 with different substrate thicknesses and types of trays. The trial was carried out using a factorial randomized design (2x2x3) with 6 replicates. The first factor was the type of trays: i.e., non-perforated and perforated tray. The second factor was the thickness of the substrate: i.e., 1.5 and 3.0 cm, while the third factor was the duration of fermentation: i.e. 5, and 7 days. The variables observed were moisture content, dry matter loss (DML), mannanase and saccharification activities, soluble protein content, their specific activities, and yield. Statistical analysis showed no interactions between the three factors, but there were interactions between types of trays and substrate thicknesses, as well as type of trays and incubation times on the mannanase activity and yield of mannanase. The results showed that DML was observed on day 7 were around 31.43- 36.89. The highest mannanase activity was observed on the non-perforated tray with 3 cm thickness on day 7. The saccharification activity towards palm kernel meal was better in the non-perforated tray on day 7 but not influenced by The yield value of mannanase and saccharification activities on a non-perforated tray with 3.0 cm thickness on day 7 was also the highest. Based on energy efficiency and the cost of production, it can be concluded that the optimum condition to produce the BS4 enzyme was observed in the non-perforated tray with 3 cm thickness and fermented for 7 days.

Biocontrol Potential of Fungal Endophytes against Fusarium oxysporum f. sp. cucumerinum Causing Wilt in Cucumber.

Endophytic fungi have received much attention as plant growth promoters as well as biological control agents against many plant pathogens. In this study, 30 endophytic fungal species, isolated from various plants in China, were evaluated using in vitro dual culture assay against Fusarium oxysporum f. sp. cucumerinum, causing wilt in cucumber. The results of the present study clearly showed that all the 30 endophytic fungal isolates were highly capable of inhibiting the mycelial colony growth of Fusarium oxysporum f. sp. cucumerinum with inhibition % over 66% as compared to control treatments. Among all of them, 5 isolates were highly effective such as, Penicillium sp., Guignardia mangiferae, Hypocrea sp., Neurospora sp., Eupenicillium javanicum, and Lasiodiplodia theobromae, respectively. The Penicillium sp. and Hypocrea sp. were highly effective as compared to other isolates. From in vitro results 10 best isolates were selected for greenhouse studies. The results of the greenhouse studies showed that among all of them 3 endophytic fungal isolates successfully suppressed wilt severity when co-inoculation with pathogen Fusarium. oxysporum f. sp. cucumerinum. The endophytic fungi also enhanced plant growth parameters of the host plants, the antagonistic fungal isolates increased over all plant height, aerial fresh, and dry weight as compared to control.

Polyketides from the endolichenic fungus Eupenicillium javanicum and their anti-inflammatory activities

Seven undescribed polyketides javanicols A−E, 5-epi-citreoviridin and 5-epi-isocitreoviridin, together with five known compounds, were isolated from the endolichenic fungus Eupenicillium javanicum. The structures of these polyketides were determined by means of extensive spectroscopic analyses, electronic circular dichroism (ECD) calculations and gauge-independent atomic orbital (GIAO) NMR shift calculations. These compounds were evaluated for potential anti-inflammatory activity against LPS-activated RAW 264.7 cells. Javanicol E and (+)-terrein displayed moderate inhibitory effects on NO production, with IC50 values of 17.00 and 13.46 μM, respectively.

Peptides with antioxidant properties identified from casein, whey, and egg albumin hydrolysates generated by two novel fungal proteases

There are many diseases linked to oxidative stress, including cancer. Importantly, endogenous antioxidants are insufficient to protect against this process. Peptides derived from food proteins produced by hydrolysis have been investigated as exogenous antioxidants. The present study aimed to identify novel peptides with antioxidant potential produced from egg and milk proteins hydrolysis with two new fungal proteases isolated from Eupenicillium javanicum and Myceliophthora thermophila. The degree of hydrolysis at several time points was calculated and correlated to DPPH scavenging and metal chelating assays, all hydrolysates presented antioxidant activity. Casein hydrolyzed by the M. thermophila protease showed the best antioxidant activity. The identified sequences showed that the proportions of amino acids that influence antioxidant activity support the antioxidant assay. Our data reveal the conditions necessary for the successful generation of antioxidant peptides using two novel fungal proteases. This opens a potential new avenue for the design and manufacture of antioxidant molecules.

Application of BS4-enzyme on the methane production from mixture of cattle manures and waste paper

<p class="abstrak2">Cellulose from abundant newspaper waste could be transformed into methane through anaerobic fermentation. This research was carried out to compare the gas production including methane between samples containing feces and waste paper mixture as inoculum and substrate, respectively and added with and without BS4 enzyme. The enzyme was produced in Indonesian Research Institute of Animal Produce (IRIAP) by growing Eupenicillium javanicum BS4 in coconut meals. There were three treatments, i.e., 30% manure (M30); 15 % manure + 15 % paper waste (MP 30); MP30 + 3 mL BS4 enzyme equal to 0.42 U/g dry matter (MPE30) The percentage of waste papers addition in feces was calculated on dry matter (DM) basis and every treatment had five replications. Total gas and methane productions were measured weekly, while dry matter losses were determined during 5 week fermentation. Interactions between treatments and incubation time were analyzed using completely randomized design each week. Kind of substrates influenced both total gas and methane productions during incubation time. Both waste papers and enzyme addition enhanced gas production. The highest total gas and methane productions for five weeks incubation were highly significantly observed (P<0.01) in MP30 and MPE30 compared to M30. Addition of enzyme significantly increased total gas and methane productions in the first week. The highest methane and total gas yield/g dry matter were obtained by BS4 enzyme addition. It was concluded that BS4 enzyme is good in accelerating and increasing the transformation efficiency of waste paper and manure mixture for biogas production.</p><p><strong>Key Words</strong>: Methane, Fibrenolytic-Enzyme, Waste Papers, Cattle Manures</p>

Analysis of the Specificity and Biochemical Characterization of Metalloproteases Isolated from Eupenicillium javanicum Using Fluorescence Resonance Energy Transfer Peptides

Enzymes have important features that may facilitate their application in industrial processes and have been used as alternatives to chemical catalysts. In particular, proteases can be isolated from microorganisms, which provide important sources of advantageous enzymes for industrial processes. For example, Eupenicillium javanicum is a filamentous fungus that has been shown to express industrially applicable enzymes and chemical components, such as antifungal compounds. The biotechnological potential of E. javanicum and proteases made us search a novel protease from this microorganism. The macromolecule was isolated, the main biochemical properties was evaluated, and the specificity of the protease subsites was determined. The protease was produced under solid-state bioprocess with wheat bran and isolated by two chromatography steps with yield of 27.5% and 12.4-fold purification. The molecular mass was estimated at 30 kDa. The N-terminal sequence of the first 20 amino acid residues was AVGAGYNASVALALEKALNN. The enzyme presented higher proteolytic activity at pH 6.0 and 60°C. The protease is stable at wide range of pH values and temperatures and in the presence of surfactants. The “primed” side of the catalytic site showed the highest catalytic efficiency of the enzyme isolated from E. javanicum. The S′1 subsite is responsible for catalyzing the protease reaction with substrates with tyrosine in P′1. These findings provide important insights into the biochemical characterization of a highly active protease from E. javanicum and may facilitate the development of industrial processes involving this protease.

Carbohydrate and protein digestion of palm kernal cake using Mannanase BS4 and papain cocktail enzymes

Supplementation of the mannanase BS4 enzyme on palm kernel cake (PKC) increased its metabolisable energy (ME), and supplementation with protease is expected to increase its protein digestibility. Therefore, the purpose of this research is to determine the optimum proportion of cocktail enzymes between BS4 β-mannanase (produced by Eupenicillium javanicum) and protease (papain) and their degradation activities on carbohydrate and protein of PKC. The β-Mannanase was produced by the mold through solid substrate fermentation (SSF) using coconut meal as the substrate. The papain was extracted from papaya latex (PL), collected by longitudinal incisions on unripe papaya fruit and oven dried overnight. The evaluation of enzyme cocktails for PKC hydrolysis was done at pH 5.8 and 40°C which are similar with poultry intestine condition and both enzymes are still active. The β- mannanase BS4 : papain were mixed with some proportions, i.e.: 100 : 0; 75 : 25; 50 : 50; 25 : 75 and 0 : 100% (by volume) in order to study the optimum cocktail composition ratio. The activities of β-mannanase towards gum locust bean was 86 U.ml -1 and papains PL activity was 18,000 U.g -1 respectively. PL was chosen for synergistic reaction and compared with a commercial Merck papain (CMP, 20,000 U.g -1 ) as positive control. β-Mannanase BS4 showed carbohydrate digestion activity, and protein digestion activity was not detected. Papain showed protein digestion activity and no carbohydrate digestion activity. Enzyme cocktails of 50 : 50 from PL protease showed slightly increased in synergistic protein digestion activity in PKC. However, its reduction sugar production was much lower than 100 : 0 and 75 : 25. Amino acids production by enzyme cocktails 75 : 25 were higher than that of 100 : 0. As a result, the best volume composition of β-mannanase BS4 and papaya latex was 75 : 25 (v:v) or 14 : 10 (U:U). Key W ords : Mannanase BS4, Papain, Cocktail-Enzymes, Palm Kernel Cake

Total Synthesis of Epoxyeujindole A.

The total synthesis of epoxyeujindole A, a structurally unusual indole diterpenoid isolated from Eupenicillium javanicum, has been accomplished for the first time. The synthesis features a late-stage cationic cyclization strategy, which took advantage of an electron-rich olefinic substrate. The CDE ring system was assembled via an enantioselective conjugate addition/alkylation, a Luche cyclization, and a Nozaki-Hiyama-Kishi reaction. The heavily substituted A ring was constructed through a Suzuki-Miyaura coupling and a cationic cyclization, and the bridged fused B ring was formed through a Prins reaction.

Evaluasi Jamur Antagonis dalam Menghambat Pertumbuhan Rigidoporus microporus Penyebab Penyakit Jamur Akar Putih pada Tanaman Karet

Rigidoporus microporus is a pathogen causing white root disease i n rubber plants , which is very difficult to control. The use of antagonistic fungi i s expected to control th is pathogen because it has the inhibitory ability through the competition, antibiosis or parasitism mechanisms. The objective of this research was to evaluate ten antagonistic fung i in inhibiting the growth of R. microporus in vitro. The research was conducted from January to April 2013, in the Laboratory of Plant Protection, Indonesian Industrial Beverages Crop Research Institute (IIBCRI) Sukabumi. The research consisted of: (1) the growth rate of colony, (2) inhibition , (3) secondary metabolites, and (3) antagonistic mechanism. Each test using a completely randomized design . The antagonist fungi used are : Trichoderma virens, T. hamatum, T. amazonicum, Eupenicillium javanicum, Penicillium simplicissimum, P. citrinum, P. pinophilum, Paecilomyces lilacinus, Aspergillus fijiensis , and Hypocrea atroviridis (= T. atroviride ) , in which each of these fungi repeated 3 times. The results showed that the Trichoderma genus have inhibitory capability against R. microporus stronger than the others. Four genus of Trichoderma fungi have better competition mechanism against R. microporus compared to Penicillium , Eupenicillium, Paecilomyces , and Aspergillus . In addition, T. virens and H. atroviridis also have parasitism ability, while P. lilacinus and E. javanicum have antibiosis mechanism against R. microporus . The antagonistic fung i which were most potential to be developed as a biological control for white root disease , i.e.: T. virens, T. hamatum, and H. atroviridis .

Analyzing the relation between the microbial diversity of DaQu and the turbidity spoilage of traditional Chinese vinegar

Vinegar is a traditional fermented condiment, and the microbial diversity of DaQu makes the quality of vinegar products. Recently, turbidity spoilage of vinegar sharply tampered with the quality of vinegar. In this study, the relation between the microbial diversity of DaQu and turbidity spoilage of vinegar was analyzed by plating technique, PCR-denaturing gradient gel electrophoresis (DGGE), and high-performance liquid chromatography (HPLC). The 16S rRNA sequencing and DGGE analysis indicated that Bacillus (Bacillus subtilis, Bacillus amyloliquefaciens, and Bacillus thuringiensis) and Lactobacillus (including Lactobacillus acidipiscis and Lactobacillus pobuzihii) species were the dominant contaminants in vinegar products. Meanwhile, DGGE analysis showed that the dominant bacteria in DaQu belonged to genera Bacillus, Lactobacillus, Pseudomonas, Weissella, Saccharopolyspora, Enterobacter, and Pantoea. However, only two yeast species (Pichia kudriavzevii and Saccharomycopsis fibuligera) and seven mold species including Aspergillus oryzae, Aspergillus niger, Aspergillus candidus, Rhizopus microspores, Eurotium herbariorum, Absidia corymbifera, and Eupenicillium javanicum were detected in the DaQu. The population level of fungi was below 5 log CFU/g in DaQu. The chemical and physical properties of vinegar and sediments were also determined. On the basis of a combined microbial diversity-chemical analysis, we demonstrated that turbidity spoilage of vinegar was a result of cooperation among the low population level and abundance of fungal species in DaQu, the suitable climate conditions, and the contaminants in vinegar. This is the first report to analyze the relation between the microbial diversity of DaQu and turbidity spoilage of vinegar.

Multivariate Analysis of the Stability of Spray-Dried Eupenicillium javanicum Peptidases

Peptidases occupy a central position in the enzyme market because of their importance in many areas, such as for physiological processes, foods, and detergents, as well as in the pharmaceutical, leather, and biotechnology industries. Microbial production is among the major sources of peptidases because it presents many advantages when compared with other methods. In this study, the metallopeptidases produced by the fungus Eupenicillium javanicum under a solid-state fermentation bioprocess were spray-dried. The enzymatic extract was dried using drying adjuvants, and optimal conditions for preserving enzymatic activity were studied following a Box-Behnken experimental design. The spray process factors studied were the air-drying temperature, enzyme feed flow rate, and the proportion of enzyme/additive. The responses analyzed were the dry extract yield and enzymatic activity after spray drying. Additionally, the stability of the dry extracts was assessed during 180 days at 4°C and 25°C. The results revealed extract yields of up to 66.12% and good enzymatic activity for intermediate values of temperature and adjuvant proportions. Furthermore, the dried enzymatic extracts showed potential for future commercial applications because of their stability at 25°C for 180 days.

Improving the Quality of Palm Kernel Cake Through Fermentation by Eupenicillium javanicum as Poultry Ration

Improving the Quality of Palm Kernel Cake Through Fermentation by Eupenicillium javanicum as Poultry Ration