Vol. 38, June, 2011.

1/38 PRODUCTION OF LIPASE ENZYME BY CLINICAL PSEUDOMONAS SPECIES ISOLATED FROM MANSOURA UNIVERSITY HOSPITALS  

Mohamed A. El-Sokkary, El-Sayed E. Habib and Shaymaa Hassan

Microbiology department, Faculty of Pharmacy, Mansoura University,

Mansoura, Egypt

The production of extracellular lipase from pseudomonas isolates collected from Mansoura University Hospitals was optimized by batch fermentation employing shake flasks. Out of 54 clinical isolates, 28 strains were lipase producers. Strain No. 45 was selected as the highest lipase producer for further studies. Maximum lipase yield (7208 U/ml) was obtained using brain heart infusion (BHI) medium with 2% tween 80 at 37°C, with shaking at 150 rpm, pH 7 and 4% bacterial inocula. Studies on the effect of mineral and vegetable oils on lipase production revealed that oils decrease the production of lipase produced by using BHI medium. In this respect paraffin and sunflower oils were the least suppressive oils. Studies on intracellular lipase showed that lipase accumulated intracellularly, but in a limited yield as compared to the extracellular lipase. Stability of lipase to temperatures up to 70°C has shown that it has moderate thermostability.

2/38 THE EFFECT OF LIQUID SEAWEED EXTRACT ON GERMINATION AND GROWTH OF RAPHANUS SATIVUS AND ERUCA SATIVA

Mlouk M. El-Kazan

Botany Department, Faculty of Science, King Abdul Aziz University, Saudi Arabia

The present study focused on the chemical composition of the liquid seaweed extract and its effect on seed germination of Raphanus sativus and Eruca sativa mill at different concentrations and different soaking time. The used seaweeds were red (Acanthophora specifera), brown (Padina pavonica and Cystosiera myrica), and green (Enteromorpha prolifera and Ulva lactuca). The mineral contents and phytohormones of the liquid seaweed extract were estimated. The liquid seaweed extracts (LSE) are rich in nitrate, phosphorous, sodium, potassium, calcium, magnesium, iron, manganese, and zinc and copper. The liquid seaweed extracts contain the growth promoting hormones Auxin, cytokinnins and gibberellins. Thus, these extracts when applied to seeds, stimulate growth of the plants. The seeds soaked in low concentrations of LSE (1% red and brown seaweeds extract and 1.5% green ones) for 6 and 12 hrs recorded 100% of germination and maximum growth of shoot and root length. Among the five LSE tested, C. myrica, E. prolifera and P. pavonica exhibited the best response. The study revealed that lower concentration of LSE was beneficial for the overall growth of the crop plant.

 

 

3/38 PHYSIOLOGICAL AND COMPARATIVE STUDIES OF POLLUTION ON SOME PLANTS GROWN AT THE ROAD SIDE LOCATIONS BETWEEN AL- KOBBA AND AL-OBOUR AREA, EGYPT

Abdel Haleem M.A. Mohammed, Ali A.A. El-Mashad, Amin E. Dowidar, Laila M. Zaky and Rania S.H. Amin

Department of Biological and Geological Sciences, Faculty of Education, Ain Shams University, Cairo, Egypt.

The present investigation of some plant species was conducted to study the effect of air pollution on (Dodonaea viscosa, Bougainvillea spectabilis and Ficus nitida) during 2 seasons (winter; December and summer; July), on some metabolic changes that may be associated with growth of plant leaves exposed to air pollution at road sides between Al- Kobba and Al-Obour city (The University area, Al-ObourMarket and Al-Tagnid square). Results indicated that an increasing of all the selected heavy metals (Pb, Cd, Zn and Mn) accumulation in dust on the surface of leaves, in leaves and soils collected from Al-Tagnid square and Al-Obour market as compared with the University area in Al-Obour city at summer and winter seasons, Moreover, increases in pH of soils, minerals in leaves of plants and soils was detected. This accumulation caused a reduction in fresh and dry matter of leaves of selected plants, % of water content and area of leaves, pigments (chlorophyll a, b, carotenoids and total pigments), total carbohydrates and total proteins.  

4/38 EFFECT OF POLLUTION ON THE PHYSICO-CHEMICAL PROPERTIES OF THE SURFACE WATER AND THE BIODIVERSITY OF SEAWEEDS IN THE RED SEA AT SOUTH OF JEDDAH, SAUDI ARABIA

Adel M. El-Gendy, Hanan H. Omar* and Mlouk M. El-Kazan**

Chemistry Dept., Faculty of Education, King Abdulaziz University, Saud Arabia.

*Microbiology Dept., Faculty of Science, King Abdulaziz University, Saud Arabia.    

**Botany Dept., Faculty of Science, King Abdulaziz University, Saud Arabia.

This paper presents the results of preliminary assessment of the surface water quality along the Red Sea at south of Jeddah (Al Khumrah area), Saudi Arabia. Water and algal samples of the Red Sea were collected seasonally from three different sites. The measurements of physico-chemical parameters in the surface water were carried out. The results showed considerable variations in water quality depending upon seasons and the location. Site 1 occupied the first order of pollution followed by site 2 and 3. Sodium was the dominant element with concentrations ranged between 2.00 and 8.92 mg/l. The abundance of macroelements was found to be in the order Na+> Mg2+> Ca2+> K+. The concentrations of trace elements were high and have the following order: Na+> Mg2+> Ca2+> K+. Zn2+> Mn2+> Fe2+> Cu2+> Cr2+> Ag+> Pb2+> Cd2+. A list of 28 algal taxa was recorded in the investigated sites during the different seasons. The brown and red algaeconstitute 46.4% and 32.1%, respectively of the total number of species, whereas the green algae were the poorest taxonomic groups (21.4%). The number of the recorded taxa were 16, 21, 12 and 22 among summer, autumn, winter, and spring, respectively. This means that spring, and autumn were the best seasons for the growth of the studied seaweeds. The predominant seaweeds were the green algae (Ulva and Enteromorpha) at site 1 and 2 and the brown algae (numerous taxa of Saragassum) at site 3. The results indicated that the area has suffered a measurable pollution. The discharge of sewage is suspected to enhance the spread of the different contaminants over large areas and may represent a potential threat to the marine life and the health of the human.


5/38 METHYL ISOTHIOCYANATE FROM CALLUS CULTURES   OF CAPPARIS ORIENTALIS DUH. AND CAPPARIS LEUCOPHYLLA DC.

Ghada A. Hegazi, Abdel Monem M. Sharaf*, Saad R. Eid

Tissue Culture Unit, Plant Genetic Resources Department, Desert Research Center, Cairo, Egypt

*Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Cairo, Egypt

The production of methyl isothiocyanate from callus of two Capparis species cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of dichlorophenoxy acetic acid (2,4-D) with or without benzyl adenine (BA) and methionine concentrations was investigated. MS medium supplemented with 2 mg/L 2,4-D + 0.5 mg/L BA was the best medium for callus induction from leaf sections of both Capparis orientalis and Capparis leucophylla, it gave the maximum mean fresh weight of callus and percentage of increase in fresh weight after two subcultures. Callus cultures established from both Capparis species were shown to produce methyl isothiocyanate. Capparis leucophylla was more responsive to produce methyl isothiocyanate in vitro than Capparis orientalis. Comparing the yields of methyl isothiocyanate in callus cultures with that in the leaves of the intact wild Capparis leucophyllaplant, by HPLC analysis, showed that the precursor feeding by the amino acid methionine, at a concentration of 2 mg/L, greatly enhanced the accumulation of methyl isothiocyanate up to 12.6 times of its amount in the intact plant.

6/38 INFLUENCES OF SEAWEED EXTRACTS ON GROWTH OF SEEDLINGS OF SOME CROP PLANTS

Hanan H. Omar, Mlouk M. El-Kazan* and Adel M. El-Gendy**

Microbiology Dept., Faculty of Science, King Abdulaziz University, Saud Arabia.

*Botany Dept., Faculty of Science, King Abdulaziz University, Saud Arabia.

**Chemistry Dept., Faculty of Education, King Abdulaziz University, Saud Arabia.

The present study an attempt to investigate the effect of seaweed liquid extracts (SLE) of red alga Gracilaria corticata and green alga Enteromorpha flexuosa on growth, pigment content, carbohydrate, protein and mineral constituents of Zea mays, Helianthus annus and Vigna unguiculata seedlings after 14 days of germination. The mineral composition and phytohormones of SLE were measured. The low concentrations of SLE promoted markedly seedling growth. The shoot length, root length, chlorophyll a & b, carotenoids, carbohydrate and protein constituents of seedlings was found maximum at 0.5% and 1% SLE of G. corticata and E. flexuosa, respectively. Improved nutrient uptake (N, P, K, Ca, Mg and trace elements) was also observed with SLE application. The seaweed liquid extracts contained a higher amount of minerals and was rich in N, P, Na, Ca, Mg, K, Fe, Mn, Zn and Cu. The concentration of phytohormones, IAA, gibberellins and cytokinins, was varied in the SLE. The content of hormones in liquid extract of E. flexuosa was higher than that in G. corticata. Thus, application of seed soaking in the seaweed liquid extracts at low concentrations could be promising option for growth enhancement of plants.


7/38 OPTIMIZATION OF CYCLOSPORIN A PRODUCTION USING LOCAL ISOLATE OF FUSARIUM SOLANI NRC 105.

Hassan A. El-Menoufy, Magdi A. Amine*, Ibrahim. S. Abd El-Salam

and Shimaa. I. Abd Elmonem

Department of Natural and Microbial Products Chemistry, National Research Centre, Egypt

*Department of Microbiology and Immunology, Faculty of   Pharmacy, Cairo University, Egypt

A local isolate of Fusarium solani NRC 105 was selected among different microorganisms tested as a new CyA producing strain. Production of CyA using the free cells was investigated under different growth conditions, involving, cultivation medium, fermentation time, inoculum size, inoculum age, pH value and the addition of some amino acids. The result revealed that the best CyA volumetric production (170 mg/l) was obtained by using fermentation medium consists of (g/l): glucose, 30; bactopeptone, 10; KH2PO4, 5; KCl, 2.5; pH 6.3, inoculum size 2% at 96 h age, as well as addition of 0.2g% L-val, at 200 rpm for 8 days.

8/38 EFFECT OF IRRADIATION ON ANTIMICROBIAL AND ANTIOXIDANT ACTIVITY OF FENNEL ESSENTIAL OIL

Mona M. Abd El Mageid; Shahinaz A. Helmy and Wafaa M. Shahin*

Department of Food Sci., Fac. of Agric., Cairo Univ., Giza, Egypt.

*National Center for Radiation Research and Technology (NCRRT), Nasr City,

Cairo, Egypt.

Fennel seeds were treated with γ-irradiation at different doses (5 to 30 kGy) as a sterilizing method compared with the non-irradiated sample and stored for 12 months at room temperature. The evaluation of the treatment efficiency depends on microbial and chemical studies, beside, antimicrobial and antioxidant activities of the extracted essential oil. The results show that the untreated samples were highly contaminated with total plate count (1.5x 105 CFU/g) as well as mould and yeast counts (4.9x104 CFU/g). Gamma irradiation was more effective in decontamination, starting at 10 kGy; however, no microbial growth was detected at 15 kGy. Also, significant difference was there among the entire non irradiated and irradiated fennel for the yield of essential oil through different storage times. Twenty six compounds were identified for the non irradiated or irradiated fennel essential oils. Trans-anethole was the main component. No changes were recorded in the components % up to γ-irradiation dose of 10 kGy. Fennel essential oil exhibited antimicrobial activity against Gram-negative, Gram-positive bacteria; molds and yeasts. Significant difference could be seen among all the tested microorganisms throughout the different doses of γ-irradiation. These inhibitions of fennel essential oils could be attributed to specific essential oil compounds, especially, trans anethole. The % of DPPH scavenging activity (antioxidant activity) increased with the increasing of fennel essential oil from the concentration of 250 to 2500 ppm, that were in significant difference. Results indicated that as dose of γ-irradiation increased up to 20 kGy, the % of DPPH did not increase for most of the different tested concentration of fennel essential oils.

9/38 PRODUCTION AND PURIFICATION OF URICASE ENZYME BY BACILLUS SUBTILIS SK09 ISOLATED FROM WADY EL-NATROON SOIL.

Enyat M. Desouky, Samir S. El-Louboudy* and Amira Y. Mahfouz

Botany and Microbiology Dept. Faculty of Science (Girls), Al-Azhar University

*Botany and Microbiology Dept. Faculty of Science (Boys), Al-Azhar University

Bacillus subtilis SK09 (WN7) was selected as the most potent uricolytic bacteria which allowed to grow on uric acid - containing medium under optimal conditions. Parameters controlling uricase production were studied, where the best substrate concentration was at 0.05%(g/v), pH: 6, for 72h incubation period, at 30°C, sucrose was the best carbon source, sodium nitrate was the best nitrogen source, alanine was found to give the highest enzyme productivity among all other tested amino acids in this study, Riboflavin amended at concentration 250ppm was the best inducer for uricase production, ferric chloride at 1150ppm induces uricase production. The optimum bottle capacity for uricase production by B.subtilis SK09 was found to be 500ml. Uricase was purified using sephadex G-200 gel filtration column chromatography technique. The molecular weight of enzyme was 82 kDa where SDS-PAGE of enzyme contains one active band. Properties of purified uricase were studied where the maximum activity of uricase enzyme was obtained at 45°C, thermostability of purified enzyme was at 45°C for 5 minutes, pH:8.5, when incubated for 5 minutes with substrate concentration of 0.4% and enzyme volume of 500µl.

10/38 CHANGES OF BIOACTIVE COMPOUNDS OF STRAWBERRY FRUITS AS A RESULT OF DIFFERENT PROCESSING TECHNIQUES

Mona M. Abdel-Magied, Enayat M. Hassan and Ghena M. Abdrabo*

Cairo University, Faculty of Agric, Food Sci&Tech. Dept, Egypt

* Nutrition and Food Sci. Dept, Faculty of Home Economics, King Abd El Aziz University, KSA.

Strawberry is an excellent source of food ingredients, however improper processing could affect the product quality. Strawberry cultivars “Festival and Sweet Charlie” were used through this study. Strawberries were processed to dehydrated and Jam. The effect of processing on the chemical composition, total phenols total anthocyanin and antioxidant capacity were determined. Biological evaluation was carried out as hypocholestrolemic effect of the dehydrated strawberryies polyphenols (as 0.3 of 0.6 polyphenols). for both cultivars on the experimental rats. This study showed that the total ellagic acids content was the predominante phenolics in the two cultivars. Pelargonidin -3- glucoside was the predominate anthocyanin regarding to the phenolic compounds. Results also showed that some reduction was happened for the total phenols, total anthocyanins antioxidant activity and Vit C for both cultivars as a result of dehydration of jam processing. Cooking strawberries to make jam redused 72% of ascorbic acid.Moreovers Storage of jam up to 6 months resulted in an degradation of ascorbic acid. The total phenolic content decreased of serum LDL concentrations in rats fed diets containing 0, 3 or 0, 6 of polyphenols from the two strawberry cultivars.An improve in AST and ALT enzymes was showen through the groups of rats fed with strawberry polyphenols. These results point out the beneficial effects of pheoilc antioxidants from strawberry fruits in preventing oxidative damage in Vivo.

11/38 EXTRACTION AND PURIFICATION OF CHITINASES FROM   SOME CEREALS AND LEGUMES

Sobhy M. Mohsen, Mohamed H. Aly, *Samiha M. Abdel-Salam

and *Ghada A. Alfaumy

Food Science Department, Faculty of Agriculture, Cairo University.

**Agriculture Research Center.

Chitinases were extracted from different cereals i.e ''barley, wheat & sorghum'' and various legumes i.e ''faba bean, soy bean & chick pea''. The crude extracts showed that barley G126, wheat B4 and sorghum G15 had the highest enzyme activity. Three methods of extractions, i.e. phosphate buffer, water and NaCl were used. The highest activity was observed by using phosphate buffer comparing to the other methods. Chitinases were purified by using ammonium sulfate (30-60%w/v) for barley G126 and (20-50%w/v) for wheat B4 and sorghum G15 then fractionated on sephadex G-100. Fractions A and B were obtained from barley, wheat and sorghum samples. The optimum temperature of barley B chitinase at 50ºC, and stable at 30ºC - 50ºC for 135min. All chitinases i.e. barley A, wheat A,B and sorghum A,B have optimum temperature at 40ºC. Optimum pH of all chitinases were observed pH 6, and stability at pH 4-7. The Km of purified chitinases from barley A,B and sorghum A were found to be the same for both methods of calculations, i.e. Line weaver and Hanes being 2.2, 0.80 and 1.6 mg.ml-1, respectively. Meanwhile km of purified chitinases from sorghum B and wheat A,B were 4.16, 2.6 and 4.5mg.ml-1 by line weaver and 4.0, 2.8 and 4mg.ml-1 by Hanes respectively. Purified chitinases from barely, wheat and sorghum were tested for their Antifungal activities and minimum inhibitory concentration against pathogens and spoilage causing molds frequently found in foods. Results showed that the highest inhibition was observed on A. flavous 1 and A. flavous 2 and Candida albicans while the lowest inhibition was on A. niger 1, A. niger 2 and Rhuzopus sp.

12/38 IN VITRO PROPAGATION OF PERIPLOCA APHYLLA – AN ENDANGERED MEDICINAL PLANT

Fawzia A. Ebad, M. A. AbdAlla*, Hemaid Ebrahim Ahmidan Soliman* and

Walla Mohamed Abed Elmaksood

Botany and Microbiology Department, Faculty of Science, Al-Azhar University, (Girls Branch), Cairo, Egypt.

*Tissue Culture Unit, Genetic Resources Department, Ecology and Dry Land Agriculture Division, Desert Research Center,11753 El-Matarya St., Elmatarya, Cairo, Egypt

An efficient protocol is described for the rapid in vitro multiplication of an endangered medicinal plant, Periploca aphylla (Asclepiadaceae), via enhanced axillary bud proliferation from nodal explants collected from young healthy shoots of natural plants. Effects of different growth regulators [Benzyl adenine (BA), 2-isopentenyladenine (2iP), a-Naphthalene Acetic Acid (NAA), kinetin (KN), Zeatin (Zi), 3-Indole Butyric Acid (IBA) and Indole Acetic Acid (IAA)] with various concentrations on in vitro morphogenesis were investigated. The highest number of shoots (11.93) and the maximum average of shoot length (2.53 cm) were obtained on MS medium fortified with 3mg/l BA+ 0.5mg/l 2iP after 8 weeks. Rooting was best achieved on full-strength MS media fortified with 1mg/l IBA +0.5mg/l NAA with mean number of 5.32 of roots formed per explant and with mean length of 5.46cm of roots. Plantlets regenerated in vitro with well developed shoot and roots were successfully established in pots and grown in the greenhouse with an 89 to 92% survival rate. The regenerated plants did not show any immediate detectable phenotypic variation. The described method can be successfully employed for large-scale multiplication and conservation of P. aphylla.

13/38 PHYLOGENY, PHYSICO-CHEMICAL PROPERTIES AND STRUCTURE PREDICTION OF TRITICUM AESTIVUM HMW GLUTENIN SUBUNITS USING BIOINFORMATICS TOOLS.

Amr E. Shalaby, Mohamed M. El-Ghoul*, El-Sayed A. El-Abasawy**,

Alaa A. Hemeida** and Ali M. El-Refy.

Biotechnology Department, Faculty of Agriculture, Al-Azhar University,  

Cairo, Egypt.

*Department of Mathematics, Faculty of Science, Tanta University, Tanta, Egypt.

**Genetic Engineering and Biotechnology Research Institute, Sadat City,

Minoufiya, Egypt.

The high molecular weight (HMW) subunits are a group of proteins which are present in the grain solely as components of high molecular mass glutenin polymers. They are important in many food systems such as breadmaking and are reported to have the strongest correlation with strong dough properties. So, we have used several computational tools to firstly construct Phylogenetic tree then secondly we predict the physico-chemical properties, secondary structural features, motifs/patterns/domains, 3D tertiary structure and quaternary structure. We have built a consensus that is derived from analysis using different algorithms to predict several structural features of the Triticum aestivum HMW glutenin subunit (Dy12) (Accession ID: Q6RX93) protein (the origin of constructed phylogenetic tree). From consensus results, we found four distinict N-terminal helices (2: 19; 24: 51; 63: 74, 78:89) and one C-terminal helix (623: 635). Random coil is predicted in between the N-terminal and C-terminal. Because of the absence of known domains, we used repeat detection programs to obtain a more accurate partitioning of the sequence in terms of domains (Nt-domain- central domain- Ct- domain). 3D structure prediction tools produced eleven models. Analysis of these models produce the best two models for Nt-domain generated from Triticum aestivum alpha-amylase inhibitor (1hssa). The first model (model 11) represents the region (35-87) of Nt-domain with high accuracy. The second model (model 4) represents the region (35-141) of Nt-domain with less accuracy. Analysis of the best two models (4,11) showed that there are 4 systeines were found in the first and second model, only 2 systeines (C43, C65) form disulfide bond. This disulfide bond connects between the first 2 helices (42:50, 58:70) that forms helix–turn–helix motif. Superimposing between the second model (model 4) as a query and predicted Quaternary structure for Triticum aestivum alpha-amylase inhibitor (1hssa) as a subject showed that one region of model 4 (104-141aa) represents the interface between two molecules. there are not any systeines form disulfide bond in the interface region but there are many polar amino acids can form hydrogen bonds and also, there are two amino acids (GLU104, ARG135) can form ionic bond. Presence of these noncovalent hydrogen and salt bridge bonding between glutenin (Dy12) subunits and polymers may be important in stabilization.
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