ISSN 2736-1756
Research Article
Advanced Journal of Microbiology Research Vol. 2010
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2010 International Scholars Journals
Full Length Research Paper
Improved antimicrobial activity of the Tanzanian edible mushroom Coprinus cinereus (Schaeff) Gray by chicken manure supplemented solid sisal wastes substrates
Liberata Nyang’oso Mwita, Anthony Manoni Mshandete and Sylvester Leonard Lyantagaye*
Department of Molecular Biology and Biotechnology, College of Natural and Applied Sciences, University of Dar es Salaam, Tanzania.
Accepted 05 April, 2010
Abstract
The Tanzanian edible mushroom species Coprinus cinereus was grown on sisal waste substrates supplemented with chicken manure with the aim to evaluate the effects of the chicken manure supplement on the antimicrobial activity of the mushroom’s extracts. Crude ethyl acetate extracts were prepared from the mushroom’s fruiting bodies harvested at pre-capping, capping and post capping stages, and the extracts were tested for antimicrobial activity, using the agar well method. The antimicrobial activity was observed only in capping and post capping stages of the mushrooms and the activity generally increased with increased percentage of manure supplementation. These findings show that Tanzanian edible C. cinereus mushroom contains antimicrobial compounds and chicken manure could be used in the cultivation of the mushroom to increase the production of active secondary metabolites, which could be used as lead compounds for discovery of new and more effective drugs against microbial infections.
Key words: Coprinus cinereus, antimicrobial, chicken manure.
Liberata Nyang’oso Mwita, Anthony Manoni Mshandete and Sylvester Leonard Lyantagaye*
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2010
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2010 International Scholars Journals
Full Length Research Paper
Identification of a restriction endonuclease (SacC1) from Saccharomyces cerevisiae
Mukaram Shikara
Biotechnology Division, Applied Sciences Department, University of Technology, Baghdad, Iraq.
Accepted 21 August, 2010
Abstract
SacC1 is a novel restriction endonuclease from Saccharomyces cerevisiae that recognizes the palindromic sequence 5’CTCGAC3’ cleaving both DNA strands upstream and downstream of its recognition sequence and makes a staggered cut at the distance of five bases from the recognition sequence on the upper strand and at the seventh base on the complementary strand. It shares similar characteristics with Sac I from Streptomyces achromogenes as well as Sst1 from Streptomyces Stanford and Psp124B1 from Pseudomonas species. It has been purified by ammonium sulphate precipitation, dialysis, and gel filtration using phosphocellulose, DEAE-cellulose and Sephadex G-100 with an optimal pH range (7.5-8.5), active at 37°C and dependent on Mg +2 or Mn2+ which increases its activity by 4- and 2-folds, respectively, while other cations decrease its activity to some extents. Cleavage on both sides of the recognition sequence is characteristic of Type IIB systems but all IIB enzymes studied so far have been found to recognize discontinuous sites and a distinctive subunit/domain organization that is not present in the SacC1 enzyme. There are similarities between SacC1 and other homing endonucleases belonging to the LAGLIDADG family such as a requirement for Mg2+ (or Mn2+) for cleavage to take place, optimal activity at alkaline pH and stimulation of the reaction by moderate concentrations of the monovalent cation.
Key words: Purification, recognition site, restriction enzyme, Saccharomyces, Streptomyces, Type IIB.
Mukaram Shikara
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2010
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2010 International Scholars Journals
Full Length Research Paper
FLAG tag module for PCR based gene targeting
Rajeev Ranjan, Sumit Kumar Verma and Shakil Ahmed*
Molecular and Structural Biology Division, Central Drug Research Institute, CSIR
Chattar Manzil Palace, MG Road, Lucknow- 226001, India.
Accepted 29 February, 2010
Abstract
Epitope tagging of yeast proteins has become an efficient tool for biochemical analysis of protein of interest. The epitope-tagged proteins can be used for western blotting, immunoprecipitation and immunofluorescence experiments without the need to raise specific antibodies, thus saving considerable time and expense. We have constructed plasmid containing FLAG tag with kanMX6 module, which allows selection of G418-resistant cells in yeast. The same set of primers that amplify module constructed by Bahler et al. (1998) can be used to amplify the FLAG tag module constructed in this study. The linear DNA fragment containing FLAG tag module with flanking homology region of gene of interest can be efficiently integrated on the yeast genome, using homologous recombination. We have successfully FLAG tag wat1/pop3 gene at its chromosomal locus and confirmed by western blot analysis. This construct can be very useful for generating C terminal tagging of desired genes at its normal chromosomal locus without interfering with their function.
Key words: S. pombe, epitope tagging, FLAG tag, pFA6a plasmid, wat1/ pop3.
Rajeev Ranjan, Sumit Kumar Verma and Shakil Ahmed*
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2010
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2010 International Scholars Journals
Full Length Research Paper
Evidence of antagonistic interactions between rhizosphere and mycorrhizal fungi associated with Dendrocalamus strictus (Bamboo)
Rohit Sharma1,2*, Ram .C. Rajak1 and Akhilesh .K. Pandey1
1Department of Biological Sciences, Mycological Research Laboratory, R. D. University, Jabalpur- 482 001, Madhya Pradesh, India.
2Microbial Culture Collection, Affiliated to National Centre for Cell Science, University of Pune, Ganeshkhind, Pune- 411 007, Maharashtra, India.
Accepted 28 May, 2010
Abstract
The paper deals with interactions of some microfungal strains isolated from rhizosphere soils from three different sites with ectomycorrhizal fungus Cantharellus tropicalis mycelium grown in vitro on agar plates. The rhizospheric fungi were isolated from 3 different sites of bamboo forest and grown against Cantharellus. The cross inoculation method showed that C. tropicalis was highly active against some fungi, thus resulting in different types and strength of interactions. Overgrowth was the most common interaction (45%), followed by inhibition at distance (29%), intermingling (17%) and contact inhibition (13%). The competitive strength of the ectomycorrhizal fungus was high and only affected by some fast growing sterile mycelia, an unidentified fungus and Trichoderma viride.
Key words: Cantharellus, ectomycorrhiza, mycorrhizal systems, biological control, soil micro fungi.
Rohit Sharma*, Ram .C. Rajak and Akhilesh .K. Pandey
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2010
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2010 International Scholars Journals
Full Length Research Paper
Production cellulase by different co-culture of Aspergillus niger and Tricoderma viride from waste paper
Ali A. Juwaied, Suhad Adnan and Ahmed Abdulamier Hussain Hussain Al-Amiery*
Department of Applied Science, Biochemical Division, University of Technology, Baghdad, Iraq.
Accepted 16 April, 2010
Abstract
In Iraq, there is attempts to transfer the various industrial carbon waste to veterinary proteins depend on microorganisms by using of chemical process. Five different co-culture combinations (1:1 ratio, 1 × 106 conidia)of Aspergillus niger and Trichoderma viride, mixing of A. niger and T. viride, in 24 and 48 h old monocultures of Aspergillus similar mixing of A. in 24 and 48 h old monoculture of Trichoderma and the monocultures of both were evaluated for their potential performance of cellulases production. The study indicates that the cellulases obtained from compatible mixed cultures simultaneous mixing of both fungi have more enzyme activity as compared to their pure cultures and other combinations. The fermentation experiments were performed in solid stat fermentation (SSF). Incubation time, carbon sources and initial pH of fermentation medium was optimized with simultaneous mixed culture. It was revealed that the newspaper at pH = 5 and 40°C was the best source of carbon for the enhanced production of cellulase in the compatible mixed culture experiments after 8 days of incubation with 5.70 U/ml. Based on the reported results, it may be concluded that industrial carbon waste can be a potential substrate for production of cellulase, incorporation of co-culturing A. niger and T. viride. The aim of this work is to produce of Cellulase from waste paper and reduce the pollution.
Key words: Aspergillus niger, Tricoderma viride, cellulase, culture.
Suhad Adnan and Ahmed Abdulamier Hussain Hussain Al-Amiery*, Ali A. Juwaied
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2010
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2010 International Scholars Journals
Full Length Research Paper
A specific inhibitory protein to a restriction enzyme from Saccharomyces cerevisiae
Mukaram Shikara*
Biotechnology Division, Department of Applied Sciences, University of Technology, Baghdad, Iraq. E-mail: [email protected].
Accepted 22 May, 2010
Abstract
A specific protein inhibitor for the restriction enzyme (SacC1) has been purified from Saccharomyces cerevisiae approximately 21,000 fold and its inhibitory properties have been characterized. The isoelectric points (pI) of SacCI and its inhibitor are 9.0 and 5.22, respectively. The molecular weight of SacC1, the inhibitor and SacC1-inhibitor complex were estimated by gel filtration on a Sephadex G-100 column to be 64,000, 32,000 and 85,000, respectively. The inhibitor protein inhibits SacC1 catalytic activities efficiently, but has no effect on other restriction enzymes tested. Inhibition does not occur unless SacC1 enzyme is exposed to the inhibitor protein prior to the reaction of the enzyme with DNA. The inhibitory activity is independent of temperature. The inhibition increased linearly with the addition of inhibitor to various amounts of SacC1, up to 85% inhibition. The slope of inhibition was constant irrespective of the initial amount of SacC1 and Ki value of 3.45 x 10-12 was obtained. The inhibitor interacts strongly with SacC1 and this interaction could increase the stability of the complex, possibly manifesting itself as SacC1 decreases in the dissociation rate due to the electrostatic attraction between the two groups or the stability may increase by potentially stronger electrostatic interaction. The conformational specificity between SacC1 and its inhibitor seems to be essential for their interaction. The extremely strong affinity of the inhibitor to SacC1 is remarkable and stronger than the affinity of several restriction enzymes.
Key words: Saccharomyces cerevisiae, inhibitor, protein, restriction enzyme, yeast, purification, Ki.
Mukaram Shikara*
Page: 1 - 10