LYCOPENE PRODUCTION IN MYCOBACTERIUM SMEGMATIS BY EXPRESSION OF CRT GENES FROM MYCOBACTERIUM AURUM AND PROTECTIVE EFFECT OF LYCOPENE IN VIVO AND IN VITRO AGAINST UV RADIATION

Objective: The aim of the present study was to express in Mycobacterium smegmatis the clustered mycobacterial genes coding for lycopene synthesis and to investigate the protective properties of lycopene against ultraviolet (UV) irradiation.Methods: The genes, which encode the biogenesis of lycopene in Mycobacterium aurum A+, were introduced into Mycobacterium smegmatis by electroporation. The pigments produced were analyzed by thin layer chromatography, and the absorption spectra were determined. A survival test using UV irradiations was also performed.Results: The transformed Mycobacterium smegmatis were found to synthesize lycopene with important yield (1.41± 3.09 mg/g) and was more resistant to ultraviolet irradiation than non-pigmented strain (p<0.01). Furthermore, cells of M. smegmatis not transformed but coated with lycopene are more resistant to UV than those uncoated (p<0.01).Conclusion: M. smegmatis can form orange colonies on agar plates when it is transformed with the lycopene genes, and the transformants produces 1.41 mg/g (dry weight) of this carotene. Our findings strongly suggest that lycopene has antioxidant activities and prevent the lethal action of UV irradiation on bacterial cells in vivo and in vitro, and deserves further studies considering the amelioration of the production

Response of Escherichia coli containing mycobacterial carotene genes to UV radiation

The plasmid pC5, which encodes biogenesis of lycopene in Mycobacterium aurum A(+), was partially digested by restriction endonucleases and generated fragments were cloned. After transformation of Escherichia coli (colorless bacteria) with the plasmids so constructed, seven orange clones were detected and found to carry the same recombinant plasmid (pC51). E. coli cells containing this plasmid synthesize neurosporene and lycopene, and were more resistant to ultraviolet irradiation than nonpigmented strain

Influence du Procede de Fabrication sur la Conformite aux Normes des Produits d’une Boyauderie Industrielle au Centre du Maroc

L'industrie de boyauderie a connu un développement prodigieux ces derniers temps au Maroc. Cette situation exige une conformité des produits de boyauderie destinés à l'exportation, étant donné que les produits de charcuterie sont souvent soumis à de multiples sources de contamination liées à la longueur et à la complexité de leur cheminement, c'est-à-dire, de l'étable à la table. Afin d'évaluer la qualité hygiénique des boyaux, des analyses microbiologiques ont été réalisées sur 7 lots différents de boyaux, à des stades différents de traitement dans une unité de boyauderie, au centre du Maroc. Les résultats montrent que tous les échantillons analysés durant les étapes de mesurage, égouttage et salage ont été conformes. Cependant, les étapes du calibrage, tubage et conditionnement ont présenté un pourcentage de non- conformité de 28,6, 42,9 et 14,3 %, respectivement. La non-conformité des échantillons analysés a été liée principalement à la présence de coliformes totaux et fécaux. Cette contamination fécal a sans doute été causée par le non-respect des règles d'hygiène au niveau des ateliers de production et/ou par les manipulateurs et pourrait être à l'origine d'un risque sanitaire. L'évaluation du microbisme bactérien a révélé que le calibrage, le tubage et le conditionnement sont des étapes critiques, dont la maîtrise est nécessaire pour assurer la conformité du produit fini. Ce qui traduit la nécessité d'instaurer un système de contrôle qualité.Mots clés : Boyaux, qualité hygiénique, analyses, risque sanitaire, conformité, Maro

Addressing the Challenge of Cultivars Identification and Authentication in Mediterranean Olive Collections: A Case Study in Morocco

Conservation and use of well-characterized olive (Olea europaea L.) genetic resources are the key to future olive improvement and sustainable production. Yet, authentication of plant materials in ex-situ olive collections throughout the world has received little attention. Here we characterized 95 accessions, from a collection maintained in the experimental station of INRAMeknes, Morocco, by comparing their SSR (14 markers) and morphological (11 endocarp traits) profiles to an international reference dataset with 672 distinct genotypes corresponding to 535 well-described olive cultivars from the two Worldwide Olive Germplasm Banks of Marrakech, Morocco, and Cordoba, Spain (WOGB-M/C). Results revealed 122 alleles in the Meknes collection versus 265 in the reference database, but the difference was not significant. Additionally, forty cultivars were identified in Meknes collection, among which 33 were present in the reference database. Principal Coordinates Analysis revealed that these varieties span the range of all of the 535 varieties in the international database, indicating important genetic diversity within the investigated plant materials. Finally, cases of mislabeling errors, synonyms, and redundant genotypes pertaining mainly to “Picholine marocaine” and “Frantoio” varieties have been encountered in Meknes collection. Overall, our work highlights the power of coupling modern genetic and morphological tools along with exploring reference databases for authenticating genetic cultivars in olive tree collections. &nbsp

Isolation and identification of a Staphylococcus warneri strain with anti-mycobacterial activity

Tuberculosis is the principal cause of death from infection in the world. The resurgence of tuberculosis and the increase in mycobacterial infections, as well as multidrug-resistance of mycobacteria to available antibiotics, has incentivized research on new antimycobacterial agents. Therefore, research based on water and soil samples from the Moroccan biotopes, has led to the isolation of a bacterial strain capable of inhibiting mycobacterial growth (Mycobacterium smegmatis and Mycobacterium aurum A+). The effect was due to an active substance secreted into the culture medium. Sequencing of the 16S rRNA gene identified the strain as belonging to the species Staphylococcus warneri. The active substance precipitated using ammonium sulfate, maintained its inhibitory properties, which were lost when treated with proteinase K. These results indicated that the active substance was protein. Study of the activity of the metabolite revealed its effect on M. smegmatis cell wall, facilitating genomic DNA extraction.Keywords: Tuberculosis, mycobacteria, anti-mycobacterial agents, Staphylococcus warneri, DNA extraction.African Journal of Biotechnology Vol. 12(42), pp. 611

Isolation and identification of Bacillus strains with antimycobacterial activity

Tuberculosis is the principal cause of death worldwide due to an infectious disease. The resurgence of tuberculosis, followed by the increase in prevalence of infections caused by nontuberculous mycobacteria (NTM), as well as the multi-drug resistance of mycobacteria to the majority of currently available antibiotics, have encouraged research for new antimycobacterial agents. Soil and water samples from different Moroccan biotopes, have led to the isolation of four bacterial strains (M, R, G and S), showing an inhibitory effect on mycobacterial growth. This effect was shown to be due to secreted substances in the growth medium. From subsequent analysis it was concluded that these strains produced different active substances. Sequencing of the 16S rRNA showed that these isolates belong to the genus Bacillus. The active substance from isolate M, showed the more important inhibitory effect on mycobacterial growth. It is precipitated with ammonium sulfate and lost all activity when treated with Proteinase K, revealing its protein nature

Biochemical characterization of a thermoactive and thermostable lipase from a newly isolated Trichosporon coremiiforme strain

Nonstop demand for greatly thermostable and thermoactive active lipase encourages the research for the new enzyme sources. In this study, a strain of Trichosporon coremiiforme was isolated from the traditional tannery in the city of Fez in Morocco, lipase production and their lipolytic activity was studied. Pure T. coremiiforme lipase (TCL) was obtained after ammonium sulfate fractionation, G-75 gel filtration and cation exchanger chromatography (Mono-S), having a molecular weight of 67 kDa. TCL presents a maximal activity at pH 8 and 50°C. After a 5 min treatment at 80°C, the enzyme maintained 50% of its activity, which is so far as is known. TCL previously characterized is found to be stable between pH 5 and 10 after 60 min incubation. TCL hydrolyses the long chains triacylglycerols more efficiently than the short ones. A specific activity of 1800 U/mg was measured on tributyrin or olive oil emulsion as substrate. This newly isolated lipase can be considered as a good candidature for industrial and biotechnological applications.Keywords: Trichosporon coremiiforme, lipase, purification, thermoactiveAfrican Journal of Biotechnology Vol. 12(28), pp. 4503-451

In vitro and intracellular antimycobacterial activity of a Bacillus pumilus strain

Despite the declaration of tuberculosis (TB) as a global emergency by the world health organization (WHO) about 20 years ago, the worldwide problem of this disease has worsened due to increased drug resistance of tuberculosis bacilli and acquired immune deficiency syndrome (AIDS) pandemic. Consequently, fight against multidrug and extensively drug-resistant TB is a high priority for public health and research. The present work describes the isolation of a Bacillus pumilus strain secreting a metabolite of protein nature capable of inhibiting mycobacterial growth (Mycobacterium smegmatis, Mycobacterium aurum and Mycobacterium bovis BCG). This metabolite is not toxic, accumulates within the macrophage and inactivates the bacilli with a comparable efficiency to that of the pure commercial antimycobacterial substance Amikacin