Evaluation of the Native Killer Yeasts against the Postharvest Phytopathogenic mould of Balady Orange Fruits

Yeasts are some of the most important postharvest biocontrol agents (BCAs). Postharvest oranges frequently deteriorate due to green and blue moulds, leading to significant economic losses. The purposes of the present study were to isolate blue and green moulds from infected orange fruits, to assess the ability of killer yeasts isolated from healthy orange fruits and leaves from orange orchards to control blue and green moulds and to evaluate the additive effect of BCAs in combination with 2% sodium bicarbonate (SBC), 2%, sodium benzoate (SB), 2% calcium chloride, 0.2% salicylic acid (SA) or 0.5% chitosan. Among eight fungi isolated from orange fruits showing symptoms of green and blue mulds infection, two were identified as P. digitatum and P. italicum and selected for in vitro assays. Twenty eight yeast isolates were obtained from orange leaves and from the surface of fruits. All yeasts exhibited high killer activity. Twelve yeasts reduced 22.5 –70% of P. digitatum growth while seven isolates reduced 21.1- 68.5% of P. italicum growth. The most potent yeast isolates were identified as Candida pseudotropicalis, Candida salmanticensis, Candida membranifaciens and Pichia guilliermondii. Combination of the BCAs, C. pseudotropicalis, C. salmanticensis and P. guilliermondii with SBC, CaCl2 or chitosan increased their effectiveness against P. digitatum. While combination of C. pseudotropicalis, C. membranifaciens and P. guilliermondii with these natural compounds decreased their effectiveness against P. italicum. Combination of C. membranifaciens with SA increased its effectiveness against P. digitatum. Sodium benzoate has additive effect on C. pseudotropicalis against P. digitatum and C. pseudotropicalis and P. guilliermondii against P. italicum

Improvement of cellulose degradation by cloning of endo-β-1, 3-1, 4 glucanase (bgls) gene from Bacillus subtilis BTN7A strain

The aim of this study is to construct a new recombinant strain able to degrade cellulose efficiently. The endo-β-1, 3-1, 4 glucanase (bgls) gene was cloned from Bacillus subtilis BTN7A strain by using PCR technique. The specific primers of bgls gene were deduced. Optimization of PCR mixture and program were identified. The nucleotide sequence of bgls was placed in the public domain (GenBank accession number KM009051.1). The obtained bgls DNA was cloned with pGEM®-T Easy Vector. The recombinant plasmid designated as Bgls-NRC-1 was transformed into E. coli DH5α. The successful cloning of the bgls gene was tested either by PCR or by evaluating its expression in its new bacterial host. The bgls gene was expressed efficiently in E. coli and the enzyme activity of the transformant was compared to the enzyme activity of the donor bacterial strain. The new constructs produce much higher enzyme yields than the donor bacterial strain, they produce about 29% and about 57% higher cellulase specific activity at 37 °C and 55 °C respectively. Optimization of cellulolytic activity of the new recombinant strain were described. The effect of minimal medium supplemented with CMC or cellulose, or complete medium (LB) on bgls expression were tested, the order of cellulase activity production was CMC27.2 > cellulose 21.9 > LB 19.8 U/mg protein, respectively at 24 h. CMC was proved to be the best medium for cellulase production. Results also showed that double the initial inoculum resulted in more cellulase activities in all media. Keywords: Bacillus subtilis, Endo-β-1, 3-1, 4 glucanase, PCR, Gene cloning, Gene expression, Cellulolytic activit

Optimization and molecular identification of novel cellulose degrading bacteria isolated from Egyptian environment

Cellulase producing bacteria were isolated from both soil and ward poultry, using CMC (carboxymethylcellulose) agar medium and screened by iodine method. Cellulase activity of the isolated bacteria was determined by DNS (dinitrosalicylic) acid method. The highly cellulolytic isolates (BTN7A, BTN7B, BMS4 and SA5) were identified on the basis of Gram staining, morphological cultural characteristics, and biochemical tests. They were also identified with 16S rDNA analysis. The phylogenetic analysis of their 16S rDNA sequence data showed that BTN7B has 99% similarity with Anoxybacillus flavithermus, BMS4 has 99% similarity with Bacillus megaterium, SA5 has 99% homology with Bacillus amyloliquefaciens and BTN7A was 99% similar with Bacillus subtilis. Cellulase production by these strains was optimized by controlling different environmental and nutritional factors such as pH, temperature, incubation period, different volumes of media, aeration rate and carbon source. The cellulase specific activity was calculated in each case. In conclusion four highly cellulolytic bacterial strains were isolated and identified and the optimum conditions for each one for cellulase production were determined. These strains could be used for converting plant waste to more useful compounds

Anticancer Activities of Newly Synthesized Chiral Macrocyclic Heptapeptide Candidates

As important cancer therapeutic agents, macrocyclic peptides have recently drawn great attention, mainly because they are synthetically accessible and have lower toxicity towards normal cells. In the present work, we synthesized newly macrocyclic pyridoheptapeptide derivatives. The synthesized derivatives were characterized using standard chemical and spectroscopic analytical techniques, and their anticancer activities against human breast and hepatocellular cancer cells were investigated. Results showed that compounds 1a and 1b were the most effective against hepatocellular (HepG2) and breast (MCF-7) cancer cell lines, respectively

Design, Synthesis and Docking Studies of Novel Macrocyclic Pentapeptides as Anticancer Multi-Targeted Kinase Inhibitors

A series of macrocyclic pyrido-pentapeptide candidates 2–6 were synthesized by using N,N-bis-[1-carboxy-2-(benzyl)]-2,6-(diaminocarbonyl)pyridine 1a,b as starting material. Structures of the newly synthesized compounds were established by IR, 1H and 13C-NMR, and MS spectral data and elemental analysis. The in-vitro cytotoxicity activity was investigated for all compounds against MCF-7 and HepG-2 cell lines and the majority of the compounds showed potent anticancer activity against the tested cell lines in comparison with the reference drugs. Out of the macrocyclic pyrido-pentapeptide based compounds, 5c showed encouraging inhibitory activity on MCF-7 and HepG-2 cell lines with IC50 values 9.41 ± 1.25 and 7.53 ± 1.33 μM, respectively. Interestingly, 5c also demonstrated multitarget profile and excellent inhibitory activity towards VEGFR-2, CDK-2 and PDGFRβ kinases. Furthermore, molecular modeling studies of the compound 5c revealed its possible binding modes into the active sites of those kinases