Naphthalene and crude oil degradation by biosurfactant producing Streptomyces spp. isolated from Mitidja plain soil (North of Algeria)

Petroleum and naphthalene (example of PAHs) degrading Streptomyces spp. isolates AB1, AH4, and AM2 were recovered from surface soils at Mitidja plain (North of Algeria). The degradation efficiencies were examined by HPLC and GC–MS analysis and the results showed that the biosurfactant producing isolates AB1, AH4 and AM2 could remove 82.36%, 85.23% and 81.03% of naphthalene after 12 days of incubation, respectively. During naphthalene degradation, a slight decrease in pH values was recorded for the three studied strains. Degradation metabolites were identified using GC–MS analysis of ethyl acetate extracts of the cell free-culture. The metabolism of degradation proceeds via the phthalic acid pathway for the three strains. Moreover, the selected strains showed an important degradation of the aliphatic fraction present in crude oil after 30 days of incubation. The finding suggests that the selected strains are suitable candidates for practical field application for effective in situ bioremediation of hydrocarbon-contaminated sites.     

Micromorphological features of Pleurotus pulmonarius (Fr.) Quel. and P. ostreatus (Jacq.) P. Kumm. strains in pure and binary culture with yeasts

Micromorphology of oyster mushrooms Pleurotus ostreatus (Jacq.) Quel. and P. pulmonarius (Fr.) Quel. was studied in pure and binary culture with yeasts (Cryptococcus laurentii 1629, Rhodotorula minuta 2790, Sporidiobolus salmonicolor (see text for symbol) 31A-11, Candida krusie 3452, Pichia holstii 3438). The cultures were cultivated on malt-agar and water agar. Various mycelial structures were described: strands, rings, thin searching mycelium, clamps, crystals, head-like offshoots, mycelial fragments, chlamydospores, and coralloid hyphae. Vegetative mycelia interact in different ways (forming anastomoses, strands, system of thin anucleate hyphae) within the same culture. Head-like offshoots of mycelial cells, previously regarded as spores of asexual reproduction, appeared to lack nuclei and to be filled with polyphosphates. Coralloid hyphae, which induce yeast cell lysis after direct contact, were detected only in binary culture with yeasts under condition of nitrogen deficit. The same way of feeding is typical for carnivorous mushrooms.     

Effect of culture conditions of Pleurotus ostreatus (Fr.) Kumm. on cellulases complexes activity in post-cultivated substrates

The aim of the presented research was to settle conditions and parameters of Pleurotus ostreatus (Fr.) Kumm. culture and to determine activity and yield of cellulase complexes biosynthesised by this mushroom on different substrates. Wheat straw sterilized by evaporation at temperature 58-60 degrees C during 48 hours was the first substrate. Wheat straw with Reynoutria japonica (Houtt) in 1:1 proportion, sterilized by irradiation (15 kGy), was the second, two-component, substrate. According to the obtained results it can be ascertained that P. ostreatus (Fr.) Kumm. grows and yields well on all kinds of substrates biosynthesising active cellulase complexes E.C. 3.2.1.4. glucohydrolases, both endo- and exogluconase, and E.C. 3.2.1.21. beta-glucosidase.     

Variation of Pleurotus ostreatus (Jacq. Ex Fr.) P. Kumm. (1871) performance subjected to differentdoses and timings of nano-urea

Supplementation of the growing substrate by nitrogenous additives has been known to improve the production of oyster mushroom (Pleurotus ostreatus (Jacq. ex Fr.) P. Kumm. (1871)). However, the application of nano-additives has not been reported in such cultivation yet. The study investigated the effect of nano-urea added in two different doses (3 g and 5 g per kg substrate), once (at spawning or after first flush) or twice (at spawning and after first flush) to the growing substrate consisting of wheat straw and spent oyster substrate (1:1, w/w). Results showed that the application of nano-urea once has induced the highest number of mushroom flushes (four flushes) despite the dose applied. Contrarily to early findings, where high doses of nitrogen have caused inhibition of mushroom growth and production, nano-urea application has had better effects when applied twice. With 5 g/kg, it induced the shortest period between the first and the third flush (15 days). With 3 g/kg, it resulted in the highest biological and economic yields at the third flush (332.7 g/bag and 283.1 g/bag respectively), in total (973.4 g/bag and 854.0 g/bag respectively), the highest biological efficiency (109.6%), and pileus diameter/stipe length ratio (2.8). Experimental findings of the current study may be potentially applied at commercial scale.     

Mycoremediation of crude oil contaminated soil by specific fungi isolated from Dhahran in Saudi Arabia

Crude oil biodegrading microorganism considers the key role for environmental preserving. In this investigation, crude oil biodegrading fungal strains have been isolated in polluted soil of crude-oil at khurais oil ground in Kingdom of Saudi Arabia. Among of 22 fungal isolates, only three isolates reflected potential capability for oil degradation. These isolates were identified and submitted to GenBank as (A1) Aspergillus polyporicola (MT448790), (A2) Aspergillus spelaeus (MT448791) and (A3) Aspergillus niger (MT459302) through internal-transcribed spacer-regions (ITS1&ITS2) for sequencing in molecular marker. Comparing with controls, strain (A1) Aspergillus niger was superior for biodegradation ability (58%) comparing with Aspergillus polyporicola and Aspergillus spelaeus degrading were showed 47 and 51% respectively. Employed CO₂ evolution as indicator for petroleum oil biodegradation by the fungal isolates reflected that, Aspergillus niger emission highest CO2 (28.6%) comparing with Aspergillus spelaeus and Aspergillus polyporicola which showed 13% and 12.4% respectively. capability of Aspergillus sp. to tolerate and adapted oil pollutants with successful growth rate on them, indicated that it can be employed as mycoremediation agent for recovering restoring ecosystem when contaminated by crude oil.     

Biological Decolourization of Textile and Paper Effluents by Pleurotus florida and Agaricus bisporus (White-rot Basidiomycetes)

Summary The extracellular ligninolytic enzymes of white-rot fungi are thought to catalyse the initial steps during the degradation of highly complex compounds like lignin or polycyclic aromatic hydrocarbons. We studied the ability of Pleurotus florida isolated from the foothills of the Western Ghats, India to decolourize the three dyestuffs, Reactive Green, Yellow and Blue, which are widely used in the textile industry around Coimbatore, Tamil Nadu, India. The crude culture filtrate of Pleurotus florida when incubated with different concentrations of dye decolourized it efficiently on the third day. The highest colour removal was found in the case of Reactive Blue. However, when Agaricus bisporus extract was supplemented with Pleurotus florida filtrate, the efficiency increased. The dye decolourization was advanced to the second day and the efficiency of dye decolourization of Reactive Yellow was 89% followed by Reactive Green, which was 45% when a dye concentration of 0.5% was used. Pleurotus florida filtrate alone and in combination with Agaricus bisporus extract reduced the aromatic compounds in textile and paper industry effluents on the first day with >90% efficiency.     

Bioemulsifier production by a halothermophilic Bacillus strain with potential applications in microbially enhanced oil recovery

A halothermotolerant Gram-positive spore-forming bacterium was isolated from petroleum reservoirs in Iran and identified as Bacillus licheniformis sp. strain ACO1 by phenotypic characterization and 16S rRNA analysis. It showed a high capacity for bioemulsifier production and grew up to 60°C with NaCl at 180 g l⁻¹. The optimum NaCl concentration, pH and temperature for bioemulsifier production were 4% (w/v), 8.0, and 45°C, respectively. Although ACO1 did not utilize hydrocarbons, it had a high emulsifying activity (E ₂₄ = 65 ± 5%) on different hydrophobic substrates. Emulsification was optimal while growing on yeast extract as the sole carbon source and NaNO₃ as the nitrogen source. The efficiency of the residual oil recovery increased by 22% after in situ growth of B. licheniformis ACO1 in a sand-pack model saturated with liquid paraffin.     

Cocultivation of <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> (Jacq.) P. Kumm. with yeasts

Cocultivation of Pleurotus ostreatus with eight yeast species were investigated on water agar. Special mycelial structures contacting with yeast cells were found in such cultures: nipple-like appendages and coralioid hyphae. Three out of eight species, Hanseniaspora uvarum, Rhodotorula minuta, and Saccharomyces cerevisiae were identified as trophic preferendum for P. ostreatus. These three yeast species were used for mushroom cultivation on sunflower seed peel. The biomass of fruiting bodies increased by 52.8–75.7% with the H. uvarum and S. cerevisiae suspension presence in the substrate.     

Lignocellulose degradation by Pleurotus ostreatus in the presence of cadmium

Activities of cellulolytic and hemicellulolytic enzymes endo-1,4-beta-glucanase, exo-1,4-beta-glucanase, 1,4-beta-glucosidase, endo-1,4-beta-xylanase, 1,4-beta-xylosidase and 1,4-beta-mannosidase and ligninolytic enzymes Mn-peroxidase and laccase were detected during the growth of the white-rot fungus Pleurotus ostreatus on wheat straw in the presence and absence of cadmium. The loss of substrate dry weight and Mn-peroxidase activity decreased with increasing Cd concentration, whereas the activities of endo-1,4-beta-glucanase, 1,4-beta-glucosidase and laccase were highly increased in the presence of metal. The onset of hemicellulose-degrading enzyme activity was delayed in the presence of cadmium. The degradation of a model synthetic dye Poly B-411 did not correspond to the activities of ligninolytic enzymes. This is the first report about 1,4-beta-mannosidase in P. ostreatus.     

Effect of rhamnolipid on biodegradation of hydrocarbons in non-aqueous-phase liquid (NAPL)

Aliphatic and aromatic hydrocarbons are environmental pollutants of serious concern. Their bioavailability is the major limiting factor that makes the bioremediation process slow. Therefore, the present study focuses on biodegradation of non-aqueous-phase liquids (NAPL) by a halophilic consortium (Pseudomonas aeruginosa and Escherichia fergusonii) in presence of rhamnolipid as well as a rhamnolipid-producing Pseudomonas aeruginosa AMB AS7. The study was performed in microcosms, and the residual hydrocarbons after degradation were estimated by gas chromatography. It was found that the degradation of hydrocarbons in NAPL was more in presence of rhamnolipid in comparison with their biotic controls. However, among NAPL, the degradation of phenanthrene (37.5%) and octadecane (47.8%) was found to be more by co-culture of halophilic consortium and rhamnolipid-producing P. aeruginosa AMB AS7. Denaturing gradient gel electrophoresis was performed to determine the viability of different bacterial strains (halophilic and rhamnolipid-producing bacterial strain). Besides, the results also revealed that during NAPL degradation, the cell surface hydrophobicity (CSH) of halophilic consortium increased from 9.12% to 69.55% when added with 100 mg/L of rhamnolipid, whereas CSH of rhamnolipid-producing P. aeruginosa AMB AS7 was constant at 31.9%, even though it produced about 271.8 mg/L of rhamnolipid.     

Biochemical and kinetic evaluation of lipase and biosurfactant assisted ex novo synthesis of microbial oil for biodiesel production by Yarrowia lipolytica utilizing chicken tallow

The present study explores the production of biodiesel, a sustainable replacement for depleting fossil fuel by utilizing microbial oil, which was procured from Yarrowia lipolytica employing chicken tallow as the carbon substrate. Chicken tallow, yeast extract, and MgSO₄·7H₂O were screened for biomass production through Plackett–Burman design. Further, Box–Behnken design analysis was performed, and the optimal concentration of the medium variables was found to be 20 g/L of chicken tallow, 7.0 g/L of yeast extract, and 0.45 g/L of MgSO₄·7H₂O.The various parameters viz., pH (6), temperature (30 °C), RPM (150), inoculum volume (5%, v/v), and C/N ratio (100) were optimized for maximal biomass and lipid yield, and lipid content. Nile red-stained cells were observed for intracellular lipid bodies using fluorescence microscopy, and its fluorescence intensity was measured bythe flow cytometer. The dimorphic transition and substrate assimilation of Y. lipolytica were analyzed using scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FT-IR). Batch kinetic studies revealed the concomitant synthesis of microbial lipid (4.16 g/L), lipase (43 U/mL), and biosurfactant (1.41 g/L). The GC-MS analysis of microbial oil presented the fatty acid profile as oleic acid (49.15%), palmitic acid (29.83%), stearic acid (11.43%), linoleic acid (3.83%), palmitoleic acid (3.77%), and myristic acid (1.32%).     

Evolution of the Tyrosinase Related Gene (TYRL) in Primates

Tyrosinase is the major enzyme responsible for the formation of melanin pigment and is found throughout the animal kingdom. In humans, the tyrosinase gene (TYR) maps to the long arm of chromosome 11 at band q14→q21, while a tyrosinase related gene (TYRL) maps to the short arm of chromosome 11 at pll.2°Cen. We and others have found that the TYRL locus contains sequences that are similar to exons IV and V of the authentic tyrosinase gene but lacks sequences of exons I, II, and III. In an attempt to understand the evolution of the human tyrosinase gene, we have analyzed TYR and TYRL in primates and have found that exons IV and V of the chimpanzee and gorilla TYR are very similar to the human, with the gorilla sequence being more similar than the chimpanzee. We have also found that the gorilla but not the chimpanzee contains a TYRL locus similar to the human TYRL locus.     

拳卷地钱挥发油成分分析

采用水蒸气蒸馏法从拳卷地钱中提取挥发油成分,并用气相色谱-质谱法(GC—MS)联机分析,共分 离出25个峰,鉴定了其中10种物质,占挥发油总组分的40．0％。在所分离的化合物中,碳氢化合物5种,烃 类含氧衍生物5种。主要成分为norpinane和hedycaryol,另外8种含量均在1％以上。在气相色谱分析中, 选择了合适的色谱条件,采用非极性的HP5柱,对地钱挥发油中的弱极性和非极性成分有较好的分离效果。     

Oxidation of glycerol by 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) in the presence of laccase

Glycerol has the potential of being a low-cost and extremely versatile building block. However, current transformation strategies such based on noble-metal-catalysts show several disadvantages including catalyst deactivation or negative environmental impacts. In this study glycerol was oxidized by 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) in the presence of laccase from Trametes hirsuta. Analysis of the reaction production indicated sequential oxidation to glyceraldehyde, glyceric acid and tartronic acid, finally resulting in mesoxalic acid. The number and nature of oxidation products was depended on the concentration of TEMPO used. At lower TEMPO concentrations (<6 mM) the major initial reaction product was glyceraldehyde while at higher concentration in addition considerable amounts of glyceric acid were formed. Glycerol oxidation was also shown with laccase immobilised on alumina pellets which increased laccase stability.     

Diversity in capsule and seed morphological and phytochemical features and essential oil composition of korarima (Aframomum corrorima (braun) P. C. M. jansen) collections from Ethiopia

Korarima (Aframomum corrorima (Braun) P. C. M. Jansen) is one of the Ethiopian native perennial aromatic herbs belonging to the family Zingiberaceae. Conservation of genotypes and genetic improvement of korarima has been hampered due to lack of evidences on genetic diversity of korarima. Genetic diversity of 44 korarima collections was assessed based on joint capsules and seeds morphological and phytochemical characters. Chemical composition of essential oil in seed samples has also been investigated. Physical measurements were done manually while total organic nutrients, crude fiber and total ash in the seeds were done in terms of proximate analysis. Essential oil in the seed samples was extracted by hydro-distillation and its chemical composition was analyzed with gas chromatography and mass spectrometry (GC-MS). ANOVA revealed significant variations in all the variables considered except capsule and seed shape. Ayda collections showed the largest record for fresh capsule weight (25.3 g), dry capsule weight (6.6 g), hundred seed weight (2.8 g), seed to husk ratio (2.1), total organic matter (66.6%) and seed essential oil (7.3%). Collection from Bazet was also excellent in dry weight of seed per capsule (4.45 g) while those from Metser were best in fruit husk essential oil (1.45%). Cluster analysis of the 44 collections revealed four groups each showing large (23.82–280.13) and significant (p < 0.01) pairwise genetic distance. The pattern of grouping is weak and does not match with the region of collections indicating the presence of different genetic background within the collections as well as genetic similarity among the collections. Principal component analysis (PCA) revealed about 81.2% of the total variations in the first four PCs and the pattern of grouping is similar to cluster analysis. In Essential oil composition, Bona Kike collection was remarkable in 1, 8-Cineole (60.81%). Ayda collection showed the highest m-Mentha-1,8-diene, (+)- (10.05%) and β-farnesene (24.03%). Adele Bise collection was highest in D-Limonene (20.21%). The genetic variability in essential oil composition among the samples was revealed because of spatial separation. The result suggests the potential of Ethiopian korarima in general and the collections in particular for further improvement and commercialization. Overall, the higher extent of morphological and biochemical genetic diversity observed signpost the high opportunity for genetic improvement of the crop via hybridization and selection.     

Production of biomass and ligninolytic enzymes by Pleurotus ostreatus in submerged culture.

The production of biomass and ligninolytic enzymes by Pleurotus ostreatus was analysed in synthetic medium with yeast extract and different glucose concentrations (0.5 - 20 g/l), at different pH (3.5-6.5) and incubation temperatures (23-32 degrees C). The best culture condition were: initial glucose concentration of 5 g/l, initial pH between 5.5-6.5 and incubation temperature between 26-29 degrees C. The saturation constant for glucose (Ks) was 1.75 g/l. The biomass concentration reached 8.6 g/l with a glucose addition of 20.0 g/l to the culture medium. The control of pH allowed an increment of 0.5 g/l of biomass concentration. The birreactor produced pellets with a homogeneous distribution of diameter size of 3.4 -/+ 0.2 mm. Approximately, 307 U/l of laccase and 0.41 U/l of manganese peroxidase were obtained in extracellular liquid medium and 0.015 U/g of laccase and 0.809 U/g of manganese peroxidase were obtained in solid substrate. Lignin peroxidase activity was not detected at any condition.     

Rapid micropropagation of Geodorum densiflorum (Lam) Schltr. in liquid culture

Technique for rapid mass propagation of Geodorum densiflorum (Lam)Schltr. has been developed by using thin sections of stems of in vitro regenerated plantlets as explant source. Thin sections of stems (1.0-1.5 mm) when cultured in modified liquid and semisolid Knudson C (KnC) medium produced 1.8 and 1.2 protocorm like bodies (PLBs) per explant respectively. Peptone (2 g l-1) was effective in promoting the survival percentage of the explants but had no effect on PLB production. BAP and NAA when used individually enhanced the rate of PLB production. But a significant and manifold increase in PLB production was noted when BAP (3 mg l-1) and NAA (0.5 mg l-1) in combination were added to peptone supplemented liquid and semisolid KnC medium. PLBs thus obtained were subcultured in semisolid KnC medium and obtained well developed plantlets within 10-12 weeks.