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J P Séguin

Publications and source records attributed to J P Séguin.

5 recordsLinked to original sources

1H-NMR on line monitoring of water activity during lipase catalysed esterification.

1H-NMR spectroscopy is used to determine simultaneously the water activity (aw) and the time course of an esterification reaction catalysed by a lipase. Chemical shifts signals of hydroxylic hydrogens in fast exchange (i.e the average hydroxylic signal of acid, alcohol and water) varies with water activity and ester content. Calibration curves have been established from model media composed of the substrates and various ester contents, at different water activities, in order to mimic a reaction medium. One relationship is established between water activity, hydroxylic hydrogen signal chemical shift and ester content. In order to estimate the water activity evolution as a function of time, this last relationship is applied to the hydroxylic hydrogen chemical shift measured in a reaction medium where the Rhizomucor miehei lipase in a powder form is suspended in the liquid substrates. This alternative way of determining the water activity based on hydroxylic hydrogen chemical shift presents some advantages over more classical means, i.e. time saved and inaccuracies avoided by monitoring without handling the sample.

1-Butanol↗

Identification of glucuronan lyase from a mutant strain of Rhizobium meliloti.

The Rhizobium meliloti M5N1CS (NCIMB 40472) mutant strain wich induces nodule formation on alfalfa roots, produces a (1 --> 4)-beta-D-glucuronan partially acetylated. During fermentation under specific conditions, the molecular weight of the polymer decrease, the presence of polysaccharide degrading enzyme was suspected. A glucuronan lyase was identified, this new bacterial lyase produces d.p. 4 oligoglucuronans, substituents (acetates) present on the substrate reduced the enzyme activity.

Acetylation↗

Study of parameters affecting poly(3-hydroxybutyrate) quantification by gas chromatography.

Poly(3-hydroxybutyrate) (PHB) quantification has been developed mostly using acidic methanolysis followed by GC analysis of the 3-hydroxybutyrate methyl ester. However, under our experimental conditions, only 62% of the ester was detected by GC analysis. Following the study of the different steps involved in this method (i.e., hydrolysis, esterification, and recovery of the ester), the recovery was shown to be limiting. Addition of water to the organic phase, required for its purification before injection, led to the partition of the ester between the organic and the aqueous phase. The influence of the length of acidic methanolysis time on the amount of ester detected was also investigated. NMR analysis was used to show that secondary products were absent in both phases, regardless of heating time. Moreover, increasing acid concentration and the use of lyophilized cells were shown to lead to the decrease of the treatment time. Concerning internal standard choice, methyl benzoate was found to meet all the requirements to correct injection volume errors or to follow organic phase volume changes as a function of acid and water concentrations. The validity of the method was checked on Rhizobium meliloti M5N1 cells, which are shown to produce about 60% PHB (w/w) when cultivated with fructose as the carbon source.

Chromatography, Gas↗

Effects of salts on production and on O-acetylation of glucuronan excreted by the Rhizobium meliloti M5N1 CS strain.

Acetylation determined by 1H-nuclear magnetic resonance spectroscopy and production of the glucuronan excreted by the Rhizobium meliloti M5N1CS strain during cultivation in RCS medium with and without added magnesium salts have been studied. These salts induce an increase in the degree of substitution and the molar ratio of 2,3-di-O-acetyl residues. A decrease in production is observed after 75 h of fermentation as the magnesium salt concentration increases. The presence of manganese and sodium salts in the culture induces inhibition of exopolysaccharide (EPS) production. However, the structure of the EPS is similar to that of the EPS produced by standard fermentation, without modification in the degree of substitution.

Acetylation↗

Cyclic (1-->2)-beta-D-glucans excreted by the glucuronan-producing strain Rhizobium meliloti M5N1CS (NCIMB 40472) and by the succinoglycan-producing strain Rhizobium meliloti M5N1.

During fermentation, the mutant strain Rhizobium meliloti M5N1CS, which induces nodule formation on alfalfa roots, produces a partially acetylated (1-->4)-beta-D-glucuronan. In addition to this exopolysaccharide of high molecular weight, the mutant strain produces oligoglucoronates and cyclic (1-->2)-beta-D-glucans with degrees of polymerization from 17 to 30. Under the conditions applied, magnesium has no effect on cyclic glucan production by the mutant strain, but the succinoglycan production by the wild-type strain Rhizobium meliloti M5N1 increases.

Acetylation↗