Cette thèse s'inscrit dans le cadre de la bioinformatique. Les outils mathématiques les plus utilisés dans ce travail relèvent de la théorie des graphes, des statistiques, de la théorie des ensembles et des mathématiques discrètes. Ces mathématiques ont permis de développer des modèles de systèmes biologiques ainsi que des algorithmes efficaces dans l'étude concrète de ces modèles. La nécessité d'analyses de jeux de données de très grande taille a rendu critique dans notre démarche cette notion d'efficacité des algorithmes. Il faut enfin remarquer que le champ biologique qui a servi de support à cette thèse nous a conduit à explorer un domaine particulier au sein de la théorie de la complexité, à savoir le développement et l'analyse des algorithmes d'énumération [etc...] / In the results presented in the present manuscripts, graph theory and combinatorial optimizationtecniques, have been used to model and solve biological problems. The manuscript is divided in twoparts, each one containing the mathematical and biological background of a given application and ouroriginal contributions to it.Part I groups a set of results designed for phylogenetics analysis, and in particular for reconstructingthe co-evolution of two groups of organisms (the so called co-phylogeny reconstruction problem).Although the addressed problem was treated in the available there was no method that solved suchproblem in a complete and efficient way. We thus developed and implemented a new one, calledEucalypt, with this purpose in mind. This not only provides a novel and usable software for cophylogenyreconstruction but also allows to investigate how the event-based model performs inpractice in terms of thenumber and quality of the solutions obtained. We compared our method to the available software. Bylooking at the results obtained, some interesting considerations about the advantages anddisadvantages of the commonly accepted mathematical model could be drawn. Finally, we introduceda new version of the problem where the host-switches are distance bounded: the k-bounded-All-MPRproblem. Eucalypt solves both problems in polynomial delay. These results have been accepted forpublication by the jounal Algorithms for Molecular Biology. The relative software is publicyavailable.Our studies show that the 'most parsimonious scenario' approach presents some limitationsthat cannot be ignored. To deal with these problems, we developed a second algorithm, called Coala,based on an approximate Bayesian computation approach for estimating the frequency of the events.The benefits of this method are twofold: it provides more confidence in the set of costs to be used in areconciliation, and it allows to estimate the frequency of the events in the cases where a reconciliationmethod cannot be applied. These results are currently under review by the jounal Systematic Biology.The relative software is publicy available.In Part 2 another set of studies is presented. Our original model for the contig scaffolding problem,and our algorithm MeDuSa, are presented and tested. Unlike traditional software, it does not rely eitheron paired-end information of sequencing reads or on a phylogenetic distance of the microorganismsused in the analysis. This drastically increases the usability of our software and, at the same time,reduces the computational time required for genome scaffolding. We show that the algorithmimplemented in MeDuSa, in most cases, is capable of producing less and longer scaffolds incomparison to commonly used scaffolders, while maintaining high accuracy and correctness of thepredicted joins. These results are currently under revision by the journal Bioinformatics.Finally, during the development of this method we encountered some pure theoretical open problemsand we decided to dedicate part of our job to their analysis. The last chapter is then dedicated to a setof problems, all related to the Implicit Hitting set enumeration problem. After some formal definitions,an original NP-completeness result is presented and the future directions of our work are described

Porifera (sponge) has been considered the earliest branching group of the metazoan crown, it plays an important role of evolution from protist to multicellular organisms. The sponges do not have tissues and organs. There are 15,000 species of sponges in the world. They contain a rich variety of secondary metabolites which may have the potential of becoming anticancer or antivirus drugs. The morphological characteristics of sponges may be affected by the environmental conditions and cause ambiguity and confusion in sponge identification. The complete mitochondrial DNAs of four Demospongiae sponges, Terpios hoshinota, Xestospongia testudinaria, Petrosia corticata, and Suberea clavata in Green Island were determined by PCR and primer walking. The sequences can be used for evolution and phylogenetic analyses. The complete mitochondrial genomes of the four sponges contain 20498 bp, 18988 bp, 18562 bp and 19559 bp, respectively. The genomes encode 2 rRNA genes (rns, rnl), 14 protein-coding genes (atp6, atp8-9, cox1-3, cob, nad1-6, and nad4L) and 25 tRNAs. All the genes of T. hoshinota, X. testudinaria, P. corticata are transcribed on the same strand. Whereas, some of the genes (nad 4L ~ tRNA-SerUGA) of S. clavata are encoded on the complementary strand. The results showed the differences between the mitochondrial DNA sequences of X. testudinaria and the Atlantic sponge, X. muta, are very limited, therefore, they may be reclassified as the same species. Meanwhile, S. clavata and Aplysina fulva are close phylogenetically. The conflict between molecular and morphology taxonomy should be re-examined.

MESHWR / Department of Ecology and Resources Management / The presence of harmful algal blooms (HABs) and cyanobacteria toxins in drinking water sources are known to pose a great threat to humans. The main aim of this study was to use molecular technique to determine the origins of the cyanobacteria species at Musina raw water abstraction point by identifying and comparing the non-toxic and toxic cyanobacteria species in the Limpopo River and some of its tributaries based on the phylogenetic analyses of 16S rRNA gene. The Musina water treatment plant is located downstream of a weir and the Beit bridge on the Limpopo River and the raw water supply is abstracted from 22 boreholes of which 14 are along the Limpopo River and 8 boreholes are inside the Limpopo River channel. The bottom sediments samples were collected from these rivers: Limpopo, Crocodile, Mokolo, Mogalakwena, Nzhelele, Lephalale, Sand rivers (South Africa); Notwane (Botswana), Shashe River and Mzingwane River (Zimbabwe). The physical-chemical analysis of the bottom sediments showed the availability of nutrients, nitrates and phosphates, in excess of 0.5 mg/l for most the of rivers, alkaline pH and salinity in excess of 500 mg/l. Total genomic DNA were extracted from cyanobacteria species on the bottom sediments and Polymerase Chain Reaction (PCR) method was used to detect the genetic profile of the cyanobacteria species. Molecular identification of cyanobacteria was based on PCR amplification and sequencing of the 16S rRNA gene. The 16S rRNA gene was absent from sediments of the Mogalakwena and Lephalale rivers but present in all other selected rivers. The cyanotoxins detection was also based on PCR by amplification of microcystin/nodularin and cylindrospermopsin polyketide synthetase genes. Most of the samples showed no amplification of the toxin genes. While two samples showed the amplification of cylindrospermopsin polyketide synthetase gene (Sand River and Nzhelele River Next to Tshipise) and two samples showed amplification for microcystin/nodularin synthetase gene, Crocodile River and Mzingwane River. The first was the confirmation of similarity of samples from Crocodile River downstream of hartbeespoort Dam and Shashe River to Leptolyngbya boryana with 99% bootstrap confidence. The similarity of sample from Musina borehole to Sand River upstream to Alkalinema pantanalense with 98% bootstrap. Thus, the presence of toxic genes may imply the presence of toxic cyanobacteria species in the river sediments and may be hazardous to humans because rural communities and commercial farmers abstract water from Limpopo River catchment for human consumption, livestock and irrigation. The waters of the Limpopo River basin also provide drinking water to wildlife and a habitant for aquatic organisms/animals.

Thesis (PhD) - Stellenbosch University, 2011. / ENGLISH ABSTRACT: The Albertine Rift is characterized by a heterogeneous landscape which may, at least in part, drive the exceptional biodiversity found across all taxonomic levels. Notwithstanding the biodiversity and beauty of the region, large areas are poorly understood because of political instability with the inaccessibility of most of the region as a contributing factor. The majority of studies in the Albertine Rift have focussed on charismatic mega fauna, with other taxa receiving less attention. One of the taxonomically and numerically more abundant small mammal genera is the genus Praomys, an African endemic with a wide distribution range spanning most of west, central and east Africa. Four species are typically recognized from the Albertine Rift namely P. degraaffi, P. jacksoni, P. misonnei and P. verschureni. In this study I used a combination of DNA sequence data (mitochondrial control region, mitochondrial cytochrome b and 7th intron of the nuclear ß-fibrinogen gene) as well as morphometric data (traditional and geometric) to investigate the systematics of the Praomys taxa occurring in the Albertine Rift. To allow meaningful DNA assessments and in an attempt to identify potential drivers of diversifications, other Praomys species were also included from public sequence data bases for comparisons. The main focus was on P. jacksoni (the numerically most abundant taxon; also, up to 2005, all Praomys in the Albertine Rift were mostly collected as “jacksoni”) and P. degraaffi (an Albertine Rift endemic). A surprising finding was the presence of P. mutoni; this represents a range extension for this species into the Albertine Rift. Distinct evolutionary lineages were found in both P. jacksoni (confirmed by sequence data as well as morphometrics) as well as P. degraaffi (based only on sequence data; insufficient samples precluded a full morphometric investigation). These lineages (in both P. jacksoni as well as P. degraaffi) appear to be separated along a north – south gradient; however, further investigations should confirm this. To further investigate the genetic patterns at local scales across the Albertine Rift, as well as introgression between species as revealed by sequence data, a species-specific microsatellite library was developed for P. jacksoni. Twelve polymorphic markers were identified of which nine also amplified in P. degraaffi. Introgression was confirmed between the two focal species with almost 20% of the individuals analysed being jacksoni-degraaffi hybrids. This is perhaps not so surprising given that there is considerable overlap in their ranges (between ~ 1500 m a.s.l. to 2450 m a.s.l.) as well as the relative ages of the species (the divergence time between these two species were estimated at 3.8 Mya). The presence of distinct lineages within each of these species was confirmed by microsatellite analyses (these lineages diverged approcimately at same time at ca. 3.4 Mya). As suggested by sequence and morphometric data, these lineages had a largely north – south distribution but with considerable overlap in the central Albertine Rift in the vicinity of Lake Kivu. The phylogeographic patterns obtained for both focal species were not consistent with the physical barriers such as the rivers, lakes or mountains, nor were they exclusively associated with Pleistocene phenomena such as the change of the course of the rivers or uplift; rather, the lineages predate the Pleistocene and fall firmly in the Pliocene (>3 Mya). Biogeographically, the north - south location of lineages with a centrally - located contact zone could be a result of parapatric speciation due to habitat fragmentation or past climate change, followed by secondary contact. Barcoding using genetic information provides a useful tool to identify unknown taxa, cryptic diversity or where different life stages are difficult to identify. From an invasion biology perspective, it allows for the rapid identification of problem taxa against a known data base. By adopting such a barcoding approach (senso lato), the presence of three invasive rodents was confirmed in the Democratic Republic of the Congo (DRC); these are Rattus rattus (black rat), R. norvegicus (Norway rat) and Mus musculus domesticus (house mouse). A comparison with global data available for these species revealed two possible introduction pathways namely via the shipping port at Kinshasa/Matadi (with strong links to Europe) and via the slave trade routes in the east (strong links to the Arab world and the east). Of these three taxa, only R. rattus is currently documented from the DRC although the others have received mention in the gray literature. These findings draw attention to the lack of any official policy regarding biosecurity in the DRC, and argue for the development of strict control measures to prevent further introductions. / AFRIKAANSE OPSOMMING: Die Albertine Rift word gekenmerk deur 'n heterogene landskap wat kan, ten minste gedeeltelik, die uitsonderlike biodiversiteit wat oor al die taksonomiese vlakke gevind word teweeg bring. Nieteenstaande die biodiversiteit en die skoonheid van die streek, is groot gebiede onbekend as gevolg van politieke onstabiliteit met die ontoeganklikheid van meeste van die streek as 'n bydraende faktor. Die meerderheid van studies in die Albertine Rift het gefokus op die charismatiese mega fauna, met ander taxa wat minder aandag ontvang. Een van die taksonomies en numeries meer volop klein soogdier genera is die genus Praomys, 'n Afrika endemiese groep met 'n wye verspreiding wat strek oor die grootste deel van van wes-, sentraal en oos-Afrika. Vier spesies word tipies erken van die Albertine Rift naamlik P. degraaffi, P. jacksoni, P. misonnei en P. verschureni. In hierdie studie het ek 'n kombinasie van DNA volgorde data (mitochondriale beheer streek, mitochondriale sitochroom b en 7de intron van die kern ß-fibrinogeen geen) sowel as morfometriese data (tradisioneel en meetkundig) gebruik om die sistematiek van die Praomys taxa te ondersoek. Om betekenisvolle DNA aanslae toe te laat en in 'n poging om potensiële aandrywers van diversiteit te identifiseer, is ander Praomys spesies van openbare volgorde data basisse vir vergelykings ingesluit. Die hooffokus is op P. jacksoni (die numeries volopste takson, ook, tot en met 2005 is alle Praomys in die Albertine Rift meestal as "jacksoni" versamel) en P. degraaffi ('n Albertine Rift endemiese spesie). 'n Verrassende bevinding was die teenwoordigheid van P. mutoni, dit verteenwoordig' n verspreidingsuitbreiding vir hierdie spesie in die Albertine Rift. Bepaalde evolusionêre ontwikkelingslyne was in beide P. jacksoni (bevestig deur die volgorde data sowel as morfometrie) sowel as P. degraaffi (wat slegs gebaseer is op die volgorde data, onvoldoende monsters verhinder 'n volledige morfometriese ondersoek). Hierdie lyne (in beide P. jacksoni sowel as P. degraaffi) word geskei langs 'n noord - suid gradiënt, maar verdere ondersoeke moet dit bevestig. Om die genetiese patrone op plaaslike skaal oor die Albertina Rift verder te ondersoek, sowel as introgressie tussen spesies soos geopenbaar deur die volgorde data, is 'n spesie-spesifieke mikrosatelliet biblioteek ontwikkel vir P. jacksoni. Twaalf polimorfiese merkers is geïdentifiseer waarvan nege ook amplifiseer in P. degraaffi. Introgressie is bevestig tussen die twee brandpunt spesies met byna 20% van die individue wat ontleed is as jacksoni-degraaffi basters. Dit is miskien nie so verbasend gegee dat daar aansienlike oorvleueling is in hul gebiede (tussen ~ 1500 m bo seespieel tot 2450 m bo seespieel), sowel as die relatiewe ouderdomme van die spesies (die divergensie tussen hierdie twee spesies is geskat op 3,8 Mya). Die teenwoordigheid van verskillende lyne in elk van hierdie spesies is bevestig deur mikrosatelliet ontleding (hierdie lyne het gedivergeer ongeveer 3,4 Mya). Soos voorgestel deur die DNA volgorde en morfometriese data, het hierdie lyne 'n grootliks noorde – suid verspreiding, maar met 'n aansienlike oorvleueling in die sentrale Albertine Rift in die omgewing van die Kivumeer. Die filogeografiese patrone wat vir beide die brandpunt spesies gevind is nie in ooreenstemming met die fisiese struikelblokke soos die riviere, mere of berge nie, en hou ook nie uitsluitlik verband met die Pleistoseen verskynsels soos die verandering van die loop van die riviere nie; die afstammelinge is eerder veel ouer as die Pleistoseen en val binne die Plioseen (> 3 Mya). Biogeografies, die noorde – suid plasing van die lyne met 'n sentraal geleë kontak sone kan die gevolg wees van parapatriese spesiasie te danke aan habitatfragmentasie as gevolg van verandering in die klimaat, gevolg deur 'n sekondêre kontak. Strepieskodering met behulp van genetiese inligting verskaf 'n nuttige instrument om onbekend taxa, kriptiese diversiteit of waar verskillende lewensfases moeilik is om te identifiseer, te identifiseer. Vanuit 'n indringerbiologie perspektief, maak hierdie benadering dit moontlik om vinnige identifikasies van die probleem taksa teen' n bekende data basis te bekom. Deur gebruik te maak van so 'n strepieskoderingsbenadering (senso lato), is die teenwoordigheid van drie indringende knaagdiere bevestig in die Demokratiese Republiek van die Kongo (DRK), naamlik Rattus rattus (swart rot), R. norvegicus (Noorweë rot) en Mus musculus domesticus (huis muis). 'n Vergelyking met die globale data wat beskikbaar is vir hierdie spesies het aan die lig gebring dat twee moontlike betree-roetes bestaan, naamlik via die skeepshawe by Kinshasa / Matadi (met sterk skakels na Europa), en via die slawehandel roetes in die ooste (sterk skakels na die Arabiese wêreld en die ooste) . Van hierdie drie taxa, is tans slegs R. rattus van die Demokratiese Republiek van die Kongo gedokumenteer, hoewel die ander melding ontvang in die grys literatuur. Hierdie bevindinge vestig die aandag op die gebrek aan enige amptelike beleid ten opsigte van biosekuriteit in die Demokratiese Republiek van die Kongo, en argumenteer vir die ontwikkeling van streng beheermaatreëls om verdere indringerspesies te voorkom.

Best-known as the family of the cultivated flax, Linum usitatissimum L., the Linaceae is a small but ecologically diverse family of flowering plants, with approximately 250 species distributed throughout the temperate and tropical latitudes of the world. This work is an investigation of the systematics and biogeography of the family and a portion of its largest genus, Linum, using molecular phylogenetic methods. I collected DNA sequences of rbcL and matK genes from 51 species of Linaceae, representing all 14 genera, and combined them with data from 24 other families of the order Malpighiales in phylogenetic analyses. Results strongly support the monophyly of Linaceae and subfamily Linoideae in their current circumscriptions, but subfamily Hugonioideae is poorly supported. Molecular dating analysis suggests that the temperate Linoideae diversified in the Eocene or Oligocene, while tropical Hugonioideae diversified later, during the Miocene, perhaps ruling out Gondwanan vicariance as an explanation for their Pantropical distribution. Hugonia and Linum, the largest genera in their respective subfamilies, are each found have multiple segregate genera nested within them, indicating potential need for taxonomic revision of each subfamily. In Linoideae, I further investigate the phylogeny of a lineage that includes the yellow-flowered Linum sections Cathartolinum, Linopsis, and Syllinum, and the segregate genera Cliococca, Hesperolinon, Radiola, and Sclerolinon, to provide a framework in which to assess character evolution, classification, and biogeography. With data from four chloroplast markers (matK, ndhF, trnK 3’ intron, trnL-F region) and the nuclear ITS, and extensive sampling from Linum section Linopsis from Eurasia, Africa, and the Americas, Hesperolinon and Sclerolinon are shown to be related to a lineage of Central American linums including L. mexicanum Kunth and L. guatemalense Benth., while Cliococca is affiliated with South American Linum. The phylogeny supportes previous hypotheses of the evolution of some taxonomically important characters, and several well-supported lineages are identified which correspond to previously proposed taxonomic groupings. Results also provide evidence for a single trans-Atlantic disjunction and independent Old and New World colonizations of the southern hemisphere in yellow-flowered Linum, occurring during the Miocene. / text

The aim of this thesis is to use phylogenetic analyses of mitochondrial DNA to investigate the biogeography and evolutionary relationships within the freshwater shrimp family Atyidae in Australia at a nested series of scales, both geographic and systematic. At the largest scale, the relationships between Australian and Indo-West Pacific species were inferred using the two most common atyid genera in Australia, Caridina and Paratya. Most atyids are hypothesised to have colonised Australia from Southeast Asia, but Paratya may be a Gondwanan relict given its distribution. Australian Paratya all form a strong clade, with a sister relationship to species from Tasman Sea islands. Molecular clock estimates place all of the splits within Paratya after the break-up of Gondwana, with Australia being colonised once 3½-8½ million years ago. This transoceanic dispersal is conjectured to have taken place through oceanic currents because of the amphidromous life cycle of some taxa of Paratya. Caridina has a very different biogeographic history in Australia, as numerous Australian species have close evolutionary relationships with non-Australian taxa from locations throughout the region. This implies many colonisations to or from Australia over a long period, and thus highlights the surprising adeptness of freshwater shrimp in dispersal across ocean barriers and the unity of much of the region's freshwater biota. A number of potential species radiations within Australia were also identified. This agrees with patterns detected for a large number of Australian freshwater taxa, and implies a vicariant explanation due to the development of colder, dryer climates. The systematic relationships of the remaining two Australian surface genera (Caridinides, Australatya) and two subterranean genera (Parisia, Pycnisia) were also investigated. Australatya forms a strong clade with Pacific 'Atya-like' genera, and Caridinides falls within a clade containing Australian Caridina. The hypogean genera, Parisia and Pycnisia, form a strong clade in all analyses, implying an Australian subterranean speciation. The possibility of a relationship between Parisia/Pycnisia and some Australian Caridina species may have implications for the monophyly of the highly disjunct genus Parisia, as it may descend from local Caridina species and represent convergent morphologies. The common and speciose genus Caridina was used as a model taxon for analyses within Australia. At the medium scale, molecular taxonomic techniques were used to uncover cryptic species within a problematic east Australian species complex. At least five species were detected. Phylogeographic and population genetic analyses were carried out on each of these five cryptic species, which diverged from each other in the late Miocene/Pliocene. There were very large differences between the species in the scales of overall geographic distribution, intraspecific divergence and population structure. These were characterised as either: 1) species with large ranges, low intraspecific divergence, limited phylogeographic structuring (Caridina sp. D); 2) species with large ranges, high intraspecific divergence, a high level of phylogeographic structuring (sp. B); 3) species with a limited range, low intraspecific divergence, no phylogeographic structuring (sp. E); or 4) species with limited ranges, high intraspecific divergences, a high level of phylogeographic structuring (sp. A & C). These patterns reflect a combination of large-scale factors, such as landscape structure and climate change, and small-scale factors, such as species-specific tolerances to local conditions and differing dispersal capabilities. Life history variation (egg size) between species may be correlated with different dispersal abilities. Species with the smallest eggs have the least intraspecific divergence and largest distribution, while those with the biggest eggs have the most divergence and smallest distribution, with medium-sized egg species in between. At the smallest phylogeographic scale, C. sp. C from the sand dune islands of Moreton Bay in southeastern Queensland was further analysed. Two different lineages (C1, C2) were found which diverged from each other during the late Miocene/Pliocene and so are older than the current landscape in which they are found. Small-scale phylogeographic analyses within C1, C2 and a sympatric fish identified divergences dating to the Pleistocene (about 100-300 thousand years ago). This implies that ice age sea-level changes may have structured these populations, although there is little observable influence of the last glacial maximum (about 18 thousand years ago). This study has highlighted a number of taxonomic anomalies within the Atyidae. The detection of many cryptic species implies that biodiversity within freshwater invertebrates is higher than currently appreciated. The evolutionary and biogeographic relationships of Australian atyids have proved complex, with many taxa having their own individual histories. At the large Indo-Pacific scale, dispersal is most evident, but within Australia, both vicariance and dispersal have been responsible for structuring all taxa at every scale.

<p>Lignocellulosic biomass represents an important renewable resource for biofuels production. Lignocellulosic biomass is comprised of cellulose, hemicellulose and lignin. Lignocellulosics are highly recalcitrant to enzymatic degradation and due to its complex nature a range of enzymes are required to synergistically hydrolyse biomass. Many microorganisms are capable of producing these enzymes as part of their hemicellulolytic hydrolysis system(s). The aim of this study was the characterisation of a thermophilic actinobacterial isolate (ORS10), capable of producing hemicellulosic enzymes, and the cloning and characterization of a hemicellulosic enzyme produced by the isolate. Phylogenetic analyses clustered ORS10 with species of the genus Streptomyces. BLAST analysis revealed that ORS10 was most closely related to Streptomyces achromogenes (99% identity). A small-insert genomic library was constructed and a putative acetylxylan esterase (AXEase) gene, axe10, was identified. The enzyme, Axe10, has moderate similarity to &alpha / /&beta / hydrolase proteins, and contains an esterase/lipase superfamily conserved domain and a typical AXEase catalytic triad. The axe10 gene was sub-cloned into an expression vector [pET21a(+)] and a 28.7 kDa protein with demonstrated AXE activity was purified from E. coli Rosetta (DE3) pLysS. Axe10 displayed optimum activity at 37oC and pH 7.0. Despite being derived from a thermophilic Streptomyces species Axe10 was not thermostable. However, given the relative novelty of Axe10, further characterisation and assessment of this enzyme is warranted.</p>

<p>Lignocellulosic biomass represents an important renewable resource for biofuels production. Lignocellulosic biomass is comprised of cellulose, hemicellulose and lignin. Lignocellulosics are highly recalcitrant to enzymatic degradation and due to its complex nature a range of enzymes are required to synergistically hydrolyse biomass. Many microorganisms are capable of producing these enzymes as part of their hemicellulolytic hydrolysis system(s). The aim of this study was the characterisation of a thermophilic actinobacterial isolate (ORS10), capable of producing hemicellulosic enzymes, and the cloning and characterization of a hemicellulosic enzyme produced by the isolate. Phylogenetic analyses clustered ORS10 with species of the genus Streptomyces. BLAST analysis revealed that ORS10 was most closely related to Streptomyces achromogenes (99% identity). A small-insert genomic library was constructed and a putative acetylxylan esterase (AXEase) gene, axe10, was identified. The enzyme, Axe10, has moderate similarity to &alpha / /&beta / hydrolase proteins, and contains an esterase/lipase superfamily conserved domain and a typical AXEase catalytic triad. The axe10 gene was sub-cloned into an expression vector [pET21a(+)] and a 28.7 kDa protein with demonstrated AXE activity was purified from E. coli Rosetta (DE3) pLysS. Axe10 displayed optimum activity at 37oC and pH 7.0. Despite being derived from a thermophilic Streptomyces species Axe10 was not thermostable. However, given the relative novelty of Axe10, further characterisation and assessment of this enzyme is warranted.</p>

Magister Scientiae - MSc / Lignocellulosic biomass represents an important renewable resource for biofuels production. Lignocellulosic biomass is comprised of cellulose, hemicellulose and lignin. Lignocellulosics are highly recalcitrant to enzymatic degradation and due to its complex nature a range of enzymes are required to synergistically hydrolyse biomass. Many microorganisms are capable of producing these enzymes as part of their hemicellulolytic hydrolysis system(s). The aim of this study was the characterisation of a thermophilic actinobacterial isolate (ORS10), capable of producing hemicellulosic enzymes, and the cloning and characterization of a hemicellulosic enzyme produced by the isolate. Phylogenetic analyses clustered ORS10 with species of the genus Streptomyces. BLAST analysis revealed that ORS10 was most closely related to Streptomyces achromogenes (99% identity). A small-insert genomic library was constructed and a putative acetylxylan esterase (AXEase) gene, axe10, was identified. The enzyme, Axe10, has moderate similarity to &alpha;/&beta; hydrolase proteins, and contains an esterase/lipase superfamily conserved domain and a typical AXEase catalytic triad. The axe10 gene was sub-cloned into an expression vector [pET21a(+)] and a 28.7 kDa protein with demonstrated AXE activity was purified from E. coli Rosetta (DE3) pLysS. Axe10 displayed optimum activity at 37oC and pH 7.0. Despite being derived from a thermophilic Streptomyces species Axe10 was not thermostable. However, given the relative novelty of Axe10, further characterisation and assessment of this enzyme is warranted. / South Africa

Archaeological and genetic evidence have long supported the notion that the Indian subcontinent played an important role in the dispersal of modern humans out of Africa. In the present study, two Dravidian populations, namely Lingayat (N=101) and Vokkaliga (N=102) were examined using high-resolution analyses of Y-chromosome single nucleotide polymorphism (Y-SNP) and their associated seventeen short tandem repeat (STR) loci. The results revealed a prevalence of the major indigenous Indian Y-haplogroups (H, L, F* and R2), which collectively accounted for three-fourths of the Lingayat and Vokkaliga paternal gene pool. In addition, the presence of ancient lineages such as F*-M213, H*-M69 and C*-M216 suggested that modern humans reached India very early after their migration out of Africa. Finally, high haplotype diversity values at 17 Y-STR loci for Lingayat (0.9981) and Vokkaliga (0.9901) populations as well as the absence of shared haplotypes between them emphasized the importance of independent databases for forensic casework.