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Antimicrobial and chemical analyses of selected bulbine species.Mocktar, Chunderika. January 2000 (has links)
The use of plant materials for the treatment of various diseases is very common in African countries. As traditional medicine used by the rural people does not always have a proper scientific basis, research programmes have to be undertaken to evaluate their therapeutic efficacy and safety. In traditional African medicine various Bulbine species are used to treat a number of conditions including sexually transmitted diseases, wound infections, dysentery and urinary tract infections. The Bulbine species belong to the family Asphodelaceae. There are over fifty South African Bulbine species and they are mostly herbs. Their leaves are evergreen and succulent in appearance. Bulbine species have thick fleshy tuberous roots, are easy to grow, are able to withstand drought and heat and are able to grow in poor soil.
There is very little documented information on the antimicrobial activity and chemical properties of the Bulbine species. Therefore research programmes of this nature have
to be undertaken. Various Bulbine species, viz., B. natalensis Bak, B. frutescens Willd (yellow flowers), B. narcissifalia Salm Dyck, B. abyssinica A Rich and B. frutescens Willd (orange
flowers) were collected. The plants were washed with tap water, air dried and separated into the different components. Each component was cut into small pieces and immersed in methanol: dichloromethane (1:1, v/v) for extraction. The organic solvent was decanted from the plant material and evaporated under reduced pressure. The resultant crude extracts were stored in glass vials in the freezer. In addition, the roots, stems and leaves of B. natalensis and B. frutescens (yellow flowers) were extracted aqueously. The crude organic and aqueous were subjected to various tests to evaluate their
antimicrobial and cytotoxic potential. To evaluate their antibacterial activities, the Disk Diffusion and Bore Well Methods were employed. The crude extracts were tested against various pathogens implicated in wound and urinary tract infections and dysentery. In these experiments the Disk Diffusion Method produced better results than the Bore Well Method. The crude organic and aqueous extracts were found to be effective against many of the bacteria used in this study including K. pneumoniae, S. aureus, S. typhi and S. flexneri which are considered to be troublesome pathogens. The TLC bioassay was employed to evaluate the antifungal potential of the various crude extracts against Aspergillus and Penicillium and the Disk Diffusion and Bore Well methods were used to evaluate the antifungal potential of C. albicans. The Bulbine species displayed no antifungal activity against Penicillium and limited antifungal activity against Aspergillus. The two method used to evaluate the antifungal activity of. C albicans was chosen because C. albicans grows in a similar manner to bacteria on solid and liquid culture media. Only the root extracts of the two B. frutescens varieties were inhibitory to C. albicans. The Brine Shrimp Bioassay was used to ascertain the cytotoxic potential of the crude extracts. The majority of the extracts were cytotoxic at the most concentrated dilution
(i.e., dilution 1) but not cytotoxic at the lower dilutions. The only extracts that were not cytotoxic at the most concentrated dilution were the organic extract of the root of
B. frutescens (yellow flowers), the organic extract of the root of B. narcissifolia and the organic extract of the leaf of B. abyssinica. TLC and column chromatography was carried out to evaluate the chemical composition of the Bulbine species. The TLC indicate that this technique could be a valuable tool in identifying the different species in the genus Bulbine. Column chromatogram was carried out on the extract which displayed a significant amount of antibacterial activity against the bacteria used in this study. The stem extract of B.
natalensis was chosen for further analysis. The stem extract was fractitioned into different fractions but unfortunately none of the chemical component could be identified.
According to the results obtained in this study, there is considerable scope for further studies of this genus. / Thesis (M.Med.Sc.)-University of Durban-Westville, 2000.
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Pyoluteorin as a signaling molecule regulating secondary metabolite production and transport genes in Pseudomonas fluorescens Pf-5Brodhagen, Marion L. 30 June 2003 (has links)
A major factor in the ability of Pseudomonas fluorescens Pf-5 to act as a
biological control agent is its production of antibiotics, including pyoluteorin (PLT),
2,4-diacetylphloroglucinol (2,4-DAPG) and pyrrolnitrin (PRN). The data provided in
this thesis demonstrate that the presence of any of these antibiotics in the extracellular
milieu affects production of that same antibiotic, as well as others, by Pf-5. Amending
the growth medium with antibiotics had multiple effects on secondary metabolism in
Pf-5. i) PLT positively regulated its own production, ii) 2,4-DAPG positively
regulated its own production. iii) PLT suppressed 2,4-DAPG production. iv) 2,4-
DAPG inhibited PLT production. v) PLT suppressed transcription of a heterologous
ferric-pyoverdine uptake gene. vi) PRN exerted a slight inhibitory effect on PLT gene
transcription and production.
PLT autoinduction by Pf-5 was extensively characterized, and was shown to
require concentrations of exogenous PLT in the nanomolar range. These low
concentrations are comparable to those of many molecules proposed to function in
signaling roles. PLT served as a signal between distinct populations of cells within the
rhizosphere, where it prompted autoinduction by those cells. Aside from effects of Pf-
5 antibiotics on one another, I also described the positive effect of exogenous PLT on
expression of a set of transport genes flanking the PLT biosynthetic gene cluster.
Sequence data and experimental evidence suggests that these genes encode a transport
apparatus for PLT. The deduced amino acid sequences for four adjacent open reading
frames together resemble Type I secretion apparatuses, which typically function in
transport of proteins rather than secondary metabolites. The intact transporter genes
are necessary for optimal PLT production.
Taken together, the data from the studies described herein demonstrate that i)
the production of PLT by Pf-5 can affect the production of PLT by neighboring cells,
and ii) PLT and other exogenous secondary metabolites have both autoregulatory and
cross-regulatory effects in culture. Because Pf-5 derivatives engaged in PLT crossfeeding
in the rhizosphere, it is likely that cross-feeding occurs for other secondary
metabolites as well. Thus, production of an antibiotic by one cell can profoundly affect
secondary metabolism in neighboring cells occupying natural habitats. / Graduation date: 2004
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Biosynthesis studies and mutasynthesis of myxobacterial secondary metabolites /Knight, Eva W. January 1900 (has links)
Thesis (M.S.)--Oregon State University, 2007. / Printout. Includes bibliographical references (leaves 81-83). Also available on the World Wide Web.
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Pharmacognosy of the Saururaceae : microscopy of powdered drugsMallya, Prakash Vaman 01 January 1984 (has links)
Plants of the family Sauruaceae are used in folk medicine in various parts of the world. Chemistry of some plants of this family and their chemotaxonomy has been investigated in our laboratory (12).
It is very difficult to establish identity and authenticity of commercially available crude drugs and powders of the Saururaceae because a detailed, collective study of microscopic features of these plants is not available. Original papers establishing plants of this family (see later) do not include microscopic features; they describe only gross morphologic and other classical features required for systematics. They are not useful for a pharmacologist in establishing identity of commercial drugs.
Therefore a detailed microscopic examination of powdered materials prepared in the laboratory from authentic crude drugs of the Saururaceae was undertaken . The observations are collectively presented in the form of "keys" which could be used by pharmacognosists and others involved in examination of crude drugs and herbal medicinals.
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Gene mining of biosynthesis genes and biosynthetic manipulation of marine bacteria for the production of new antibiotic candidatesZarins-Tutt, Joseph Scott January 2015 (has links)
Natural product drug discovery has traditionally been the corner stone of medicine having provided cures to many of today's most common diseases. In particular, antibiotics have revolutionised healthcare and extended human lifespan. However, since their introduction into the clinic, resistance to these drugs has arisen. With the number of new antibiotics being discovered in recent years declining, and fewer drugs making it past clinical trials, we have reached the point where antibiotic resistant infections have become common place and a serious threat to health and society. There is now an urgent requirement for the discovery of new antibiotics and in particular those with unexploited mode of action. This thesis details the different areas of natural product drug development from discovery through to analogue generation. In Chapter one, the history of natural products as therapeutics is explored with a particular focus on antibiotics and how resistance arises against these agents. It outlines why the discovery of new antibiotics is so important and new methods used to facilitate this search. Chapter two follows with the development of a screening platform for antibiotic induction, using the model Streptomyces; Streptomyces coleiolor M145. A variety of culture additives are explored for their ability to induce secondary metabolism production. Chapter three then details the sampling and identification of microbes from a pseudo-marine environment and their screening for their ability to produce secondary metabolites with antibiotic properties. The second half of this thesis centres on the non-ribosomal peptide echinomycin. Collaborators Aquapharm supplied the marine derived strain AQP-4895, capable of producing echinomycin. Chapter four details the establishment of AQP-4895 culturing conditions and the shift observed in production profile. Next Chapter five looks at producing echinomycin analogues through precursor directed biosynthesis. A range of halogenated quinoxaline carboxylic acids are synthesised and fed to AQP-4895, and the respective echinomycin analogues monitored by LC-MS. Chapter Six then aims to direct biosynthesis of the halogenated analogues, using mutasynthesis. Due to the lack of genetic data available surrounding the strain, an unusual approach was taken, using iPCR to create a template for homologous recombination.
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