Bioinformatics and molecular approaches to detect NRPS genes involved in the biosynthesis of kurstakin from <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> |
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Authors: | Ahmed Abderrahmani Arthur Tapi Farida Nateche Marlène Chollet Valérie Leclère Bernard Wathelet Hocine Hacene Philippe Jacques |
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Institution: | 1.Laboratoire des Procédés Biologiques, Génie Enzymatique et Microbien, ProBioGEM, UPRES-EA 1026, Polytech’Lille/IUT A,Université Lille Nord de France-Sciences et Technologies, USTL,Villeneuve d’Ascq Cedex,France;2.Laboratoire de Biologie Cellulaire et Moléculaire, Faculté des Sciences Biologiques,Université des Sciences et de la Technologie Houari Boumediene,El Alia. Alger,Algeria;3.Unité de Chimie Biologique Industrielle,Université de Liège, Gembloux Agro-Bio Tech,Gembloux,Belgium |
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Abstract: | Degenerated primers designed for the detection by polymerase chain reaction of nonribosomal peptide synthetases (NRPS) genes
involved in the biosynthesis of lipopeptides were used on genomic DNA from a new isolate of Bacillus thuringiensis CIP 110220. Primers dedicated to surfactin and bacillomycin detection amplified sequences corresponding respectively to the
surfactin synthetase operon and to a gene belonging to a new NRPS operon identified in the genome of B. thuringiensis serovar pondicheriensis BSCG 4BA1. A bioinformatics analysis of this operon led to the prediction of an NRPS constituted
of seven modules beginning with a condensation starter domain and which could be involved in the biosynthesis of a heptalipopeptide
similar to kurstakin. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS) performed
on whole cells of B. thuringiensis CIP 110220 confirmed the production of kurstakin by this strain. The kurstakin operon was thus used to design a new set of
degenerated primers specifically to detect kurstakin genes. These primers were used to screen kurstakin producers in a collection
of nine B. thuringiensis strains isolated from different areas in Algeria and two from the Pasteur Institute collection. For eight among the 11 tested
strains, the amplified fragment matched with an operon similar to the kurstakin operon and found in the newly sequenced genome
of Bacillus cereus or B. thuringiensis serovar pulsiensis, kurstaki, and thuringiensis. Kurstakin production was detected by MALDI-ToF-MS on whole cells for six
strains. This production was compared with the spreading of the strains and their antimicrobial activity. Only the spreading
can be correlated with the kurstakin production. |
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