Biosynthetic features and properties of xylose isomerases from Arthrobacter nicotianae, Escherichia coli, and Erwinia carotovora subsp. atroseptica

The characteristics of xylose isomerase biosynthesis in the bacteria Arthrobacter nicotianae BIM B-5, Erwinia carotovora subsp atroseptica jn42xylA, and Escherichia coli HB101xylA have been studied. The bacteria produced the enzyme constitutively. Out of the carbon sources studied, D-glucose and D-xylose were most favorable for the biosynthesis of xylose isomerase in E. carotovora subsp. atroseptica, but the least appropriate in terms of the enzyme production efficiency in E. coli. Minimum and maximum levels of xylose isomerase formation in A. nicotianae were noted, respectively, during D-xylose and sucrose utilization. An addition to the D-xylose-containing nutrient medium of 0.1–1.5% D-glucose did not affect the enzyme synthesis in A. nicotianae, but suppressed it in Erwinia carotovora subsp. atroseptica (by 7% at the highest concentration) and Escherichia coli (by 63 and 75% at concentrations of 0.1 and 1.0%, respectively). The enzyme proteins produced by the bacteria exhibited the same substrate specificity and electrophoretic mobility (PAGE) as xylose isomerase A. nicotianae, although insignificant differences in the major physicochemical properties were noted.     

Screening of glucose isomerase-producing microorganisms

The ability to convert D-glucose into D-fructose was found in 14 out of 74 species of actinomycetes and bacteria tested. High intracellular glucose isomerase activity was displayed by Arthrobacter sp. and actinomycetes Streptomyces viridobrunneus, Streptomyces sp. 1 and Streptomyces sp. 32. The first showed maximal glucose-converting potential when cultured in both glucose and xylose media, while glucose isomerase activity of Streptomyces species could be found solely in medium supplemented with xylose. The ketose enzymatically formed from D-glucose was identified as D-fructose.     

Nitric oxide selectively enhances cAMP levels and electrical coupling between identified RPaD2/VD1 neurons in the CNS of Lymnaea stagnalis (L.)

The isolated CNS of the freshwater mollusc Lymnaea stagnalis was used as a model to study the role of cAMP in NO-mediated mechanisms. The NO donor, DEA/NO (10(-5)-10(-3) M) increased cAMP concentrations in the cerebral, pedal, pleural, parietal and visceral ganglia. In contrast, in the buccal ganglia the same doses of DEA/NO decreased the level of cAMP production. The NOS inhibitor, L-NNA (10(-4) M) increased cAMP concentrations in all areas of the CNS. L-arginine (1 mM), a metabolic precursor of NO, mimicked the action of the NO-donor. The coefficient of electrical coupling between two viscero-parietal peptidergic neurons (VD1/RPaD2) was enhanced by both DEA/NO (10(-4) M) and 8-Br-cAMP (10(-4) M) whereas 8-Br-cGMP (2x10(-4) M) reduced the coupling. We suggest that cAMP-dependent mechanisms are involved in neuronal NO signaling in this simpler nervous system.     

A plate method to screen for microorganisms producing xylose isomerase

A plate method was developed to screen for xylose isomerase-producing microorganisms based on the use of 2,3,5-triphenyltetrazolium as an indicator of D-xylulose, the D-xylose isomerization product. The use of this method allows microorganisms to be differentiated by the character of the enzyme synthesis (inducible or constitutive).     

Screening of glucose isomerase-producing microorganisms

The ability to convert D-glucose into D-fructose was found in 14 out of 74 species of actinomycetes and bacteria tested. High intracellular glucose isomerase activity was displayed by Arthrobacter sp. and actinomycetes Streptomyces viridobrunneus, Streptomyces sp. 1 and Streptomyces sp. 32. The first showed maximal glucose-converting potential when cultured in both glucose and xylose media, while glucose isomerase activity of Streptomyces species could be found solely in medium supplemented with xylose. The ketose enzymatically formed from D-glucose was identified as D-fructose.     

Investigation of the synthesis of xylose/glucose isomerases in sixArthrobacter sp. strains

The substrate specificity of isomerases produced by six strains ofArthrobacter sp. was studied. The role of utilizable carbon sources in controlling enzyme biosynthesis was established. All of the strains studied were found to produce xylose isomerases efficiently, converting D-xylose into D-xylulose and D-glucose into D-fructose. All but A.ureafaciens B-6 strains showed low activity toward D-ribose,Arthrobacter sp. B-5 was slightly active toward L-arabinose, andA. ureafaciens B-6 andArthrobacter sp. B-2239, toward L-rhamnose. InArthrobacter sp. B-5, the synthesis of xylose/glucose isomerase was constitutive (i.e., it was not suppressed by readily metabolizable carbon sources. The synthesis of xylose/glucose isomerase induced by D-xylose inArthrobacter sp. strains B-2239, B-2240, B-2241, and B-2242 and by D-xylose and xylitol inA. ureafaciens B-6 was suppressed by readily metabolizable carbon sources in a concentration-dependent manner. The data obtained suggest that D-xylose and/or its metabolites are involved in the regulation of xylose/glucose isomerase synthesis in theArthrobacter sp. strains B-5, B-2239, B-2240, and B-2241.     

Some Properties of the Xylose/Glucose Isomerase of Immobilized Arthrobacter sp. Cells

The study of the xylose/glucose isomerase–containing Arthrobacter sp. B-5 cells immobilized in cobalt hydroxide gel showed that immobilization increases the substrate affinity of the enzyme, its thermo- and pH-optima of action and stability, and makes the addition of stabilizing cobalt ions to the isomerization medium unnecessary.     

Biosynthetic features and properties of xylose isomerases from Arthrobacter nicotianae, Escherichia coli, and Erwinia carotovota subsp. atroseptica

The characteristics of xylose isomerase biosynthesis in the bacteria Arthrobacter nicotianae BIM B-5, Erwinia carotovota subsp atroseptica jn42xylA, and Escherichia coli HB101xylA have been studied. The bacteria formed the enzyme constitutively. Out of the carbon sources studied, D-glucose and D-xylose were most favorable for the biosynthesis of xylose isomerase in E. carotovota subsp atroseptica, but the least appropriate in terms of the enzyme production efficiency in E. coli. Minimum and maximum levels of xylose isomerase formation in A. nicotianae were noted, respectively, during D-xylose and sucrose utilization. An addition to the nutrient medium of 0.1-1.5% D-glucose (together with D-xylose) did not affect the enzyme synthesis in A. nicotianae, but suppressed it in Erwinia carotovota subsp atroseptica (by 7% at the highest concentration) and Escherichia coli (by 63 and 75% at concentrations of 0.1 and 1.0%, respectively). The enzyme proteins produced by the bacteria exhibited the same substrate specificity and electrophoretic mobility (PAGE) as xylose isomerase A. nicotianae, although insignificant differences in the major physicochemical properties were noted.