Abstract: The present invention relates to a method of creating DNA libraries that include an artificial promoter library and/or a modified ribosome binding site library and transforming bacterial host cells with the library to obtain a population of bacterial clones having a range of expression levels for a chromosomal gene of interest.

Abstract: The invention relates to optimised micro-organism strains for the biotransformation production of molecules having NADPH-consuming biosynthetic pathways. The inventive strains can be used in NADPH-consuming biotransformation methods. Said strains are characterized in that one or more NADPH-oxidizing activities are limited.

Abstract: The present invention concerns a new method for the biological preparation of a diol comprising culturing a microorganism genetically modified for the bioproduction of an aliphatic diol, wherein the microorganism comprises a metabolic pathway for the decarboxylation of a hydroxy-2-keto-aliphatic acid metabolite with an enzyme having a 2-keto acid decarboxylase activity, the product obtained from said decarboxylation step being further reduced into the corresponding aliphatic diol, and wherein the microorganism is genetically modified for the improved production of said hydroxy-2-keto-aliphatic acid metabolite. The invention also concerns a modified microorganism for the production of an aliphatic diol.

Abstract: The present invention concerns a new method combining evolution and rational design for the preparation of a strain of micro-organism for the production of acetol from a simple carbon source. The said method comprises: growing an initial strain under selection pressure in an appropriate growth medium, said initial bacterial strain comprising an attenuation of the expression of the tpiA gene and an attenuation the expression of at least one gene involved in the conversion of methylglyoxal to lactate, in order to promote evolution in said initial strain, then selecting and isolating the evolved strain having an increased acetol or 1,2-propanediol production rate, then reconstructing a functional tpiA gene in the evolved strain. The present invention also concerns the evolved strain such as obtained, that may be furthermore genetically modified in order to optimize the conversion of a simple carbon source into acetol without by-products and with the best possible yield.

Abstract: The present invention is relative to a method for producing 1,2-propanediol by fermentation, comprising: cultivating a microorganism producing 1,2-propanediol in an appropriate medium comprising a source of sucrose, and recovering the 1,2-propanediol being produced, wherein the microorganism is able to utilize sucrose as sole carbon source for the production of 1,2-propanediol. In a preferred aspect of the invention, the source of sucrose is obtained from plant biomass, and is in particular sugar cane juice.

Abstract: The present invention concerns a modified microorganism with an increased methylglyoxal reductase activity, and its use for the preparation of 1,2-propanediol and/or acetol. In particular this increased methylglyoxal reductase activity is obtained by increasing the expression of specific genes from microorganisms. This invention is also related to a method for producing 1,2-propanediol and/or acetol by fermentation of a microorganism having an increased methylglyoxal reductase activity.

Abstract: The present invention concerns a new method for the biological preparation of 2-hydroxy-isobutyrate (2-HIBA), including a fermentation method with microorganisms modified to favour production of 2-HIBA from renewable resources. The invention also concerns the modified microorganisms used in such fermentation method.

Abstract: The present invention relates to a method for producing and preparing polyglycolate (PGA) from genetically engineered organisms. More specifically, the invention relates to a method comprising two steps; 1) culturing, in a medium containing glycolic acid or not, the microorganism expressing at least one gene encoding an enzyme(s) that converts glycolate into glycolyl-CoA, and a gene encoding polyhydroxyalkanoate (PHA) synthase which uses glycolyl-CoA as a substrate, 2) recovering the polyglycolate polymer.

Abstract: This invention provides a series of low-copy number plasmids comprising restriction endonuclease recognition sites useful for cloning at least three different genes or operons, each flanked by a terminator sequence, the plasmids containing variants of glucose isomerase promoters for varying levels of protein expression. The materials and methods are useful for genetic engineering in microorganisms, especially where multiple genetic insertions are sought.

Abstract: The present invention relates to a novel polypeptide having the enzymatic activity of conversion of methylglyoxal to lactic acid in a single step (known as glyoxalase III activity), a polynucleotide having a nucleotide sequence encoding such polypeptide and uses thereof. The invention relates to the modulation of the glyoxalase III activity in a microorganism by varying the expression level of the polynucleotide coding for such polypeptide. The invention also relates to the production of commodity chemicals, especially 1,2-propanediol, acetol, and lactic acid by fermenting microorganisms wherein their glyoxalase III activity is modulated.

Abstract: The present invention provides a method for the biological production of n-butanol at high yield from a fermentable carbon source. In one aspect of the present invention, a process for the conversion of glucose to n-butanol is achieved by the use of a recombinant organism comprising a host C. acetobutylicum transformed i) to eliminate the acetate pathway ii) to eliminate the butyrate pathway iii) to eliminate the lactate pathway and iv) to eliminate the acetone pathway. In another aspect of the present invention, the hydrogen flux is decreased and the reducing power redirected to n-butanol production by interrupting the expression of the hydrogenase gene. Optionally the n-butanol produced can be eliminated during the fermentation by gas striping and further purified by distillation.

Abstract: The present invention is related to a new method for interrupting multiple DNA sequences in Clostridia, even in genes recognized to be essential for the optimal growth of Clostridii by using a counter-selectable marker that would pinpoint the cells that have lost the plasmid and acquired a modified function that permits survival without the interrupted gene. This method is easy to perform and applicable at an industrial level. This method is useful to modify several genetic loci in Clostridia in a routine manner. This method is based on a replicative vector carrying at least two marker genes.

Abstract: The invention concerns a method for preparing 1,3-propanediol from a carbon-containing substance, said method comprising a step which consists in culturing a recombinant micro-organism not producing coenzyme B12 in the absence of coenzyme B12 or one of its precursors. The invention also concerns a nucleic acid coding for a glycerol dehydratase whereof the catalytic activity is independent of the presence of coenzyme B12 or one of its precursors and a nucleic acid coding for a 1,3-propanol dehydrogenase intervening in the synthesis of 1,3-propanediol. The invention further concerns recombinant vectors and host cells comprising said nucleic acids and the polypeptides coded by the latter.

Abstract: The present invention concerns a new method combining evolution and rational design for the preparation of a strain of micro-organism for the production of 1,2-propanediol from a carbon source. The said method comprises: growing an initial strain under selection pressure in an appropriate growth medium, said initial bacterial strain comprising an attenuation of the expression of the tpiA gene and an attenuation the expression of at least one gene involved in the conversion of methylglyoxal to lactate, in order to promote evolution in said initial strain, then selecting and isolating the evolved strain having an increased 1,2 propanediol production rate, then reconstructing a functional tpiA gene in the evolved strain; The present invention also concerns the evolved strain such as obtained, that may be furthermore genetically modified in order to optimize the conversion of a carbon source into 1,2-propanediol without by-products and with the best possible yield.

Abstract: This invention concerns a microorganism useful for the production of acetol from a simple carbon source, wherein said microorganism is characterized by: an improved activity of the biosynthesis pathway from dihydroxyacetone phosphate to acetol, and an attenuated activity of the glyceraldehyde 3-phosphate dehydrogenase This invention also concerns a method for producing acetol by fermentating a microorganism according to the invention.

Abstract: Microorganism useful for the production of 1,2-propanediol from a carbon source, wherein said microorganism is characterized by: an improved activity of the biosynthesis pathway from dihydroxyacetone phosphate to 1,2-propanediol, and an attenuated activity of the glyceraldehyde 3-phosphate dehydrogenase The invention is also related to a method for producing 1,2-propanediol by fermentation with a microorganism according to the invention.

Abstract: The present invention provides a microorganism useful for biologically producing 1,3-propanediol from a fermentable carbon source at higher yield than was previously known. The complexity of the cofactor requirements necessitates the use of a whole cell catalyst for an industrial process that utilizes this reaction sequence to produce 1,3-propanediol. The invention provides a microorganism with disruptions in specified genes and alterations in the expression levels of specified genes that is useful in a higher yielding process to produce 1,3-propanediol.

Abstract: The present invention provides a method for the anaerobic production of 1,3 propanediol, by culturing a Clostridium strain in an appropriate culture medium comprising glycerol as a source of carbon, wherein said Clostridium strain does not produce substantially other products of the glycerol metabolism selected among the group consisting of: butyrate, lactate, butanol and ethanol, and recovering of 1,3-propanediol.

Abstract: The invention concerns a method for preparing 1,3-propanediol from a carbon-containing substance, said method comprising a step which consists in culturing a recombinant micro-organism not producing coenzyme B12 in the absence of coenzyme B12 or one of its precursors. The invention also concerns a nucleic acid coding for a glycerol dehydratase whereof the catalytic activity is independent of the presence of coenzyme B12 or one of its precursors and a nucleic acid coding for a 1,3-propanol dehydrogenase intervening in the synthesis of 1,3-propanediol. The invention further concerns recombinant vectors and host cells comprising said nucleic acids and the polypeptides coded by the latter.

Abstract: The invention concerns a method for preparing 1,3-propanediol from a carbon-containing substance, said method comprising a step which consists in culturing a recombinant micro-organism not producing coenzyme B12 in the absence of coenzyme B12 or one of its precursors. The invention also concerns a nucleic acid coding for a glycerol dehydratase whereof the catalytic activity is independent of the presence of coenzyme B12 or one of its precursors and a nucleic acid coding for a 1,3-propanol dehydrogenase intervening in the synthesis of 1,3-propanediol. The invention further concerns recombinant vectors and host cells comprising said nucleic acids and the polypeptides coded by the latter.