Xylitol production from xylose by two yeast strains: Sugar toleranceNolleau, V.; Preziosi-Belloy, L.; Delgenes, J.; Navarro, J.
doi: 10.1007/BF01692875pmid: N/A
The kinetics and enzymology ofd-xylose utilization are studied in micro-, semi-, and aerobic batch cultures during growth ofCandida guilliermondii andCandida parapsilosis in the presence of several initial xylose concentrations. The abilities of xylitol accumulation by these two yeast strains are high and similar, although observed under various growth conditions. WithCandida parapsilosis, optimal xylitol production yield (0.74 g/g) was obtained in microaerobiosis with 100 g/L of xylose, whereas optimal conditions to produce xylitol byCandida guilliermondii (0.69 g/g) arose from aerobiosis with 300 g/L of sugar. The different behavior of these yeasts is most probably explained by differences in the nature of the initial step of xylose metabolism: a NADPH-linked xylose reductase activity is measured with a weaker NADH-linked activity. These activities seem to be dependent on the degree of aerobiosis and on the initial xylose concentration and correlate with xylitol accumulation.
Interaction of α2-macroglobulin with alkaline proteinase from an alkalophilicBacillus sp. grown in an extraordinarily alkaline environmentYamagata, Youhei; Ichishima, Eiji
doi: 10.1007/BF01692877pmid: N/A
The interaction of α2-macroglobulin (α2M) with an alkaline serine proteinase (ALPase I) from alkalophilicBacillus sp. grown in an extraordinarily alkaline environment was investigated. Stoichiometry of the reaction showed that ALPase I bound to α2M in a molar ratio of about 2∶1. The α2M-ALPase I complex showed about 80% of the proteinase activity shown by ALPase I in the hydrolysis of succinyl-l-alanyl-l-alanyl-l-prolyl-l-phenylalanyl-4-methyl-coumaryl-7-amide (Suc-Ala-Ala-Pro-Phe-MCA) and casein. The conformational changes in the α2M molecule caused by the complex formation at pH 7.5 were determined from electron micrographs and difference spectra. The antigenic activity of the α2M-ALPase I complex with the anti-ALPase I antiserum was found to be completely abolished. Immunoelectrophoresis of the complex incubated at pH 7.5 after 48 h showed no appreciable change, and the complex was recognized as exhibiting enhanced stability at pH 7.5.
Cotransformation of lactococcin-producing, 2.0-mega dalton and erythromycin-resistant pGB 301 plasmids toLactococcus lactis subsp.lactis protoplastGupta, R.; Grover, Sunita; Batish, V.
doi: 10.1007/BF01692878pmid: N/A
Protoplasts of plasmid-freeLactococcus lactis subsp.lactis LM 0230 and PC4 strains were cotransformed successfully with the plasmid pools ofL. lactis subsp.lactis 484, a lactosefermenting (Lac+), lactococcin-producing (Lap+), lactococcin-resistant (Lapr), sucrosefermenting (Suc+) wild strain, its derivatives, and pGB 301 erythromycin resistance plasmid (Eryr) at the frequencies of 104 transformants/μg of DNA. PC4 protoplasts were transformed at slightly lower frequencies that LM 0230 protoplasts when the same plasmid combinations were used for transformation. Agarose gel electrophoresis of plasmids from three groups of transformants, namely, Lac−Lap−Eryr, Lac+Suc+Lap+LaprEryr, and Lac−Suc+Lap+ LaprLapr, confirmed that 2.0 and 65.0 megadalton (MDa) plasmids carried genes for Suc+Lap+Lapr and Lac+ phenotypes respectively. The protoplasts could be transformed with low-molecular-weight 2.0 MDa Lap plasmid at a relatively higher frequency than those with high-molecular-weight 65.0 MDa Lac plasmid. All the transformants resembled parent culture 484 in terms of lactic acid production (0.810–0.840%), milk curdling time (6 h), and lactococcin activity (7–12 mm, zone of inhibition) againstListeria monocytogenes, Salmonella typhi, andStaphylococcus aureus. The plasmids and their respective phenotypes in PC4 transformants were genetically more stable than those of LM 0230 protoplasts. The marker plasmid pGB 301 disappeared more frequently from the transformants when present in association with the lowmolecular-weight, high-copy-number 2.0 MDa plasmid, thereby suggesting the incompatibility of these two plasmids.
Biodesulfurization of organic-sulfur compoundsPurdy, Ralph; Lepo, Joe; Ward, Bailey
doi: 10.1007/BF01692879pmid: N/A
A screening assay in which dibenzothiophene (DBT) or DBT-sulfone served as the only bioavailable source of sulfur was used to obtain two new bacterial isolates, strains UM9 and UM3, that desulfurized either substrate. Strain UM9 produced the desulfurized product, 2-hydroxybiphenyl (HBP); no other identifiable desulfurized products or released sulfate or sulfite were detected. Biodesulfurization activity occurred only for growing cultures and was depressed by free sulfate. Neither isolate grew on DBT, DBT-sulfone, or HBP as sole carbon sources. Under optimized conditions of pH and temperature, strain UM9 exhibited up to 35% greater biodesulfurization of DBT-sulfone than did UM3, and both isolates also desulfurized several other organic-sulfur compounds. The kinetics and characteristics of biodesulfurization by either UM3 or UM9, tentatively identified as species ofRhodococcus, indicated mechanisms different from those reported in the literature for other bacteria.
Postprandial variations in the activity of polysaccharide-degrading enzymes of fluid- and particle-associated ruminal microbial populationsMartin, Cécile; Michalet-Doreau, Brigitte; Fonty, Gérard; Williams, Alan
doi: 10.1007/BF01692880pmid: N/A
The distribution and specific activities of polysaccharide-degrading enzymes were monitored during the postprandial period in the liquid-associated bacteria (LAB), liquid-associated protozoa (LAP), and solid-associated microbes (SBFP) isolated from ruminal contents of cattle fed a high-cereal diet. Polysaccharide depolymerase activities were highest in the SBFP and the LAP populations. The postprandial variations in the specific activity of amylase were similar within the subpopulations. However, carboxymethylcellulase and xylanase activities increased in the first 5 h after feeding in the LAP, but were highest at the end of the postprandial period in the SBFP. Glycosidases involved in the fermentation of soluble carbohydrates increased significantly immediately after feeding in the liquid-associated microbes. β-d-Glucosidase and β-d-xylosidase were most active in the SBFP and were maximal 23 h after feeding. The activities of the plant cell wall polysaccharide-degrading enzymes and glycosidases in the SBFP were inversely related to ruminal pH; however, the activity of enzymes in the liquid-associated populations were highest in the immediate post-feed period when ruminal pH was lowest.
Purification of grapevine flavescence dorée MLO (Mycoplasma-like organism) by immunoaffinitySeddas, Abdessamad; Meignoz, Raymonde; Daire, Xavier; Boudon-Padieu, Elisabeth; Caudwell, Antoine
doi: 10.1007/BF01692881pmid: N/A
To date MLO (Mycoplasma-like organism) remain non-culturable organisms and are difficult to extract in good conditions of purity and conservation from infected hosts (plants or leafhopper vectors). An immunoaffinity procedure that permits the purification of large quantities of Grapevine Flavescence dorée MLO (FD-MLO) is described, with covalently bound and oriented IgG molecules of a previously obtained anti-FD-MLO monoclonal antibody and elution of antigens in alkaline conditions. Evidence for purity and integrity of the eluted MLO is presented. The two main antigenic components detected by rabbit polyclonal antibodies to FD-MLO were shown to be different proteins and to contain different epitopes with the use of different monoclonal antibodies. DNA extracted from the purified FD-MLO fraction hybridized with an FD-MLO DNA-specific probe.
Phospholipid fatty acid ester composition ofPasteurella multocida andActinobacillus lignieresiiFuller, Cynthia; Brignac, Paul; Champlin, Franklin
doi: 10.1007/BF01692882pmid: N/A
Cell surface hydrophobicity properties vary dramatically, whereas cell envelope phospholipid composition is essentially identical among strains ofPasteurella multocida andActinobacillus lignieresii. Fatty acid ester composition of the major phospholipid fractions from cell surface hydrophobicity variants was examined to determine whether hydrophobic properties are influenced by cell envelope fatty acid content. Individual phospholipids were resolved by preparative thin-layer chromatography, and methanolysis was performed with boron trifluoride-methanol. Gas-liquid chromatographic analysis revealed the organisms to be similar qualitatively, whereas hydrophobic variants exhibited consistently, greater and more disparate C16:0+C16:1/C14:0 ratios in all fractions. Fatty acid composition of phospholipids may be related to surface hydrophobicity properties ofP. multocida variants. However, comparative data obtained forA. lignieresii revealed a degree of similarity withP. multocida that precludes use of this parameter as a means for differentiation of thesePasteurellaceae type species, thereby supporting their taxonomic relatedness.
Legionella pneumophila virulence conserved after multiple single-colony passage on agarYamamoto, Yoshimasa; Klein, Thomas; Friedman, Herman
doi: 10.1007/BF01692883pmid: N/A
Multiple passage ofLegionella pneumophila on either supplemented Mueller-Hinton or charcoal yeast extract agar by the conventional batch passing technique results in loss of virulence. In this studyL. pneumophila virulence was maintained after multiple passage on buffered charcoal yeast extract (BCYE α) agar by a single-colony transfer technique. Virulence was determined by assessing the growth ofL. pneumophila for thioglycolate-induced susceptible A/J mouse macrophages in vitro and infectivity for susceptible A/J mice in vivo. Passage of the virulentL. pneumophila, 30 times on BCYE α agar by the single-colony transfer procedure still resulted in virulence, as compared with the nonpassaged parental bacteria, both in vitro and in vivo. Lethality for susceptible, A/J mice by systemic infection was comparable for the 30th colony-passaged bacteria and the parentalL. pneumophila. These results show that theL. pneumophila phenotype associated with intracellular growth in macrophages or infectivity for susceptible mice is stable following passage by the single-colony transfer procedure on BCYE α agar.