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Effect of oxygen on lactose metabolism in lactic streptococci.

Three strains of Streptococcus lactis, two of Streptococcus cremoris, and one of Streptococcus thermophilus metabolized oxygen in the presence of added carbohydrate primarily via a closely coupled NADH oxidase/NADH peroxidase system. No buildup of the toxic intermediate H(2)O(2) was detected with the three S. lactis strains. All six strains contained significant superoxide dismutase activity and are clearly aerotolerant. Lactose- or glucose-driven oxygen consumption was biphasic, with a rapid initial rate followed by a slower secondary rate which correlated with factors affecting the in vivo activation of lactate dehydrogenase. The rate of oxygen consumption was rapid under conditions that led to a reduction in lactate dehydrogenase activity (low intracellular fructose 1,6-bisphosphate concentration). These conditions could be achieved with nongrowing cells by adding lactose at a constant but limiting rate. When the rate of lactose fermentation was limited to 5% of its maximum, nongrowing cells of S. lactis strains ML3 and ML8 carried out an essentially homoacetic fermentation under aerobic conditions. These same cells carried out the expected homolactic fermentation when presented with excess lactose under anaerobic conditions. Homoacetic fermentation leads to the generation of more energy, by substrate-level phosphorylation via acetate kinase, than the homolactic fermentation. However, it was not observed in growing cells and was restricted to slow fermentation rates with nongrowing cells.

Journal Article↗

Presence of lactose genes and insertion sequences in plasmids of minor species of the genus lactococcus.

The type strains of all known species and biovars of the Lactococcus genus were tested for the presence of plasmids, lactose genes, and insertion sequences cloned from the lactose plasmid of Lactococcus lactis subsp. lactis. Only the biovar xylosus of this subspecies is plasmid free. The lactose plasmid is present only in lactose-positive strains except in Lactococcus plantarum. The distribution of insertion sequences varies within the type strains of the Lactococcus genus.

Journal Article↗

Structures and properties of gellan polymers produced by sphingomonas paucimobilis ATCC 31461 from lactose compared with those produced from glucose and from cheese whey

The dairy industry produces large quantities of whey as a by-product of cheese production and is increasingly looking for new ways to utilize this waste product. Gellan gum is reliably produced by Sphingomonas paucimobilis in growth media containing lactose, a significant component of cheese whey, as a carbon source. We studied and compared polysaccharide biosynthesis by S. paucimobilis ATCC 31461 in media containing glucose, lactose (5 to 30 g/liter), and sweet cheese whey. We found that altering the growth medium can markedly affect the polysaccharide yield, acyl substitution level, polymer rheological properties, and susceptibility to degradation. Depression of gellan production from lactose compared with gellan production from glucose (approximately 30%) did not appear to occur at the level of synthesis of sugar nucleotides, which are the donors of monomers used for biosynthesis of the repetitive tetrasaccharide unit of gellan. The lactose-derived biopolymer had the highest total acyl content; the glucose- and whey-derived gellans had similar total acyl contents but differed markedly in their acetate and glycerate levels. Rheological studies revealed how the functionality of a gellan polysaccharide is affected by changes in the acyl substitution.

Journal Article↗

Effects of Dimethylsulfoxide on the Lactose Operon in Escherichia coli.

Fowler, Audree V. (University of California, Los Angeles), and Irving Zabin. Effects of dimethylsulfoxide on the lactose operon in Escherichia coli. J. Bacteriol. 92:353-357. 1966.-Dimethylsulfoxide (DMSO) at a concentration of 5% (v/v) in the culture medium inhibits the growth of Escherichia coli to only a slight extent, and does not affect the differential rate of synthesis of beta-galactosidase. Resting cells remain viable after shaking in the presence of 20% DMSO for 3 hr at 37 C. Both beta-galactosidase and thiogalactoside transacetylase retain almost all activity after incubation in even higher concentrations of the solvent for many hours. DMSO decreases the permeability barrier. The rate of hydrolysis of o-nitrophenyl-beta-d-galactoside (ONPG) in whole cells containing beta-galactosidase but lacking permease is increased in cells treated with 5% DMSO. Several permeaseless strains preinduced for beta-galactosidase will grow on lactose in the presence, but not in the absence, of 5% DMSO. When permeaseless strains are grown on tetrazolium-lactose-agar, the presence of 5% DMSO causes a definite but not marked shift toward the lactose-positive character.

Journal Article↗

Primary structure of the lactose permease gene from the yeast Kluyveromyces lactis. Presence of an unusual transcript structure.

The LAC12 gene of Kluyveromyces lactis codes for an inducible lactose permease. We have determined the nucleotide sequence of a DNA fragment which includes the complete LAC12 gene. The 4.7-kilobase (kb) mRNA carrying LAC12 contained two open reading frames, ORFI (1761 bases) and ORFII (1266 bases), separated by a 573-base pair noncoding region. Mung bean and exonuclease VII mapping showed that there was no splicing of the 4.7-kb transcript and thus no intron between the two open reading frames. Chromosomal disruption of ORFI with the URA3 gene destroyed lactose transport activity, suggesting that ORFI codes for a component of the permease. Disruption of ORFII and the noncoding region between the two open reading frames did not affect the lactose permease function, indicating that they do not comprise a part of the permease. We do not know if ORFII is translated, but in either case, the structure of the 4.7-kb mRNA is unusual. We discuss possible origins for it. The peptide predicted from ORFI is hydrophobic as would be expected for a membrane-bound protein. Compared with other membrane proteins, LAC12 (ORFI) protein showed sequence similarity to the human glucose and the Escherichia coli xylose-H+ and arabinose-H+ transporters. No obvious amino acid sequence similarity was found with the lactose permease of E. coli.

Amino Acid Sequence↗

Role of soy-based, lactose-free formula during treatment of acute diarrhea.

A controlled study was conducted comparing the standard method of treating hospitalized infants with acute diarrhea (limited starvation) with the initiation of "early feeding" using a soy-based, lactose-free formula in infants of an American Indian tribe 12 months of age or younger. Forty-three patients, randomly assigned to group A, were given a soy-based, lactose-free formula four hours after hospitalization, and 44 patients, randomly assigned to group B, received standard therapy (food was withheld for the first 48 hours of hospitalization). After the first 48 hours, the same soy-based, lactose-free formula was given to the group B patients. Fluid intake and output of stool, urine, and vomitus were measured until the diarrhea resolved. Overall, group A patients showed less mean stool output (121 +/- 129 (SD) mL/kg) than group B patients (299 +/- 319 mL/kg) (P less than .001). Furthermore, the duration of illness was significantly shorter in group A patients (54 +/- 28 hours v 93 +/- 56 hours) (P less than .001). It was concluded that soy-based, lactose-free formulas can be safely used during the acute phase of diarrheal illness in infants and that their use shortens the duration of illness and decreases stool output in comparison with standard therapy.

Combined Modality Therapy↗

[The effect of dietary lactose on the ileal absorption of sodium taurocholate in rats].

When Rats were fed a lactose containing diet, both the absorption rate of sodium taurocholate at the level of ileum and the contents of bile acids of the small intestine were increased. On the contrary, feeding of lactose did not modify the daily fecal excretion of bile acids. It therefore appears that dietary lactose increases the intestinal pool of bile acids by increasing their ileal absorption rate and that this effect of lactose is not subordinated to a modification of bile acid synthesis.

Animals↗

Lactose malabsorption and intolerance in Italians. Clinical implications.

Lactose malabsorption was assessed by the hydrogen breath test in 40 Italian patients with irritable bowel syndrome and 42 controls without abdominal disturbances. Sixty-five percent of patients were "low milk consumers" (0-250 ml milk per day) compared with 38% of controls (P less than 0.02). Lactose loads of 25 and 50 g caused malabsorption in 82.5 and 87.5% patients and in 55 and 62% controls, respectively (patients vs controls P less than 0.02). Malabsorption was more frequent in the "low milk consumers" group (P less than 0.05). During a four-month lactose-free diet as the only treatment 7.5% of patients became symptom-free (and remained so for a further eight-month diet), 52.5% improved, and 40% showed no change.

Adult↗

Breath hydrogen response to milk containing lactose in colicky and noncolicky infants.

In 122 healthy newborn infants, we studied the relationship between breath hydrogen (H2) production after feedings containing lactose (human milk or commercial formula) in colicky and noncolicky infants at 6 weeks and 3 months. Eighty-three infants (68%) developed colic (mild, moderate, or severe) by 2.6 +/- 1.8 weeks of age (mean +/- SD). Zero time (baseline) breath H2 values were significantly higher in colicky compared with noncolicky infants at both 6 weeks (40.6 +/- 41.4 vs 14.8 +/- 32.9 ppm) and 3 months (27.7 +/- 38.1 vs 8.5 +/- 18.2 ppm). There were significantly more positive breath H2 tests in colicky compared with noncolicky infants at 6 weeks (78% vs 36%) and 3 months (89% vs 45%). Failure to produce H2 throughout the breath H2 test was significantly more frequent in noncolicky compared with colicky infants at 6 weeks (50% vs 18%) and 3 months (43% vs 4%). These findings remained significant even when infants with mild colic (at 6 weeks and 3 months) were included in the noncolicky group. We conclude that colicky infants produce more breath H2 in the fasting state and in response to feedings containing lactose than noncolicky infants produce. This may represent increased lactose malabsorption, differences in colonic bacterial fermentation conditions, or differences in the handling of colonic gas produced.

Breath Tests↗

Hydrogen excretion upon ingestion of dairy products in lactose-intolerant male subjects: importance of the live flora.

OBJECTIVE: To assess the effects of the ingestion of milk, yoghurt (10(8) bacteria/ml), heat-treated yoghurt (<15 bacteria/ml) and two products obtained by dilution of yoghurt with heat-treated product (10(6) and 10(5) bacteria/ml) on hydrogen production and symptoms of lactose intolerance in lactose malabsorbers. DESIGN: Double-blind, randomised cross-over design. SETTING: The study was performed in the phase 1 clinical unit of OPTIMED, Nancy, France. SUBJECTS: Twenty-four male lactose malabsorbers were selected for the study. INTERVENTIONS: Hydrogen production and adverse events were followed during 8 h after ingestion of the products. RESULTS: The results clearly demonstrate that ingestion of yoghurt with 10(8) bacteria/ml leads to lower H(2) excretion and complaints than the other products. Results observed with the products containing a reduced population of live flora remain better than those observed with milk. CONCLUSIONS: The importance of a high population of the live flora is underlined.

Adult↗

Beta-galactosidase production by Streptococcus thermophilus is higher in the small intestine than in the caecum of human-microbiota-associated mice after lactose supplementation.

Transit kinetics and survival rates of a bacterial species from yoghurt (i.e. Streptococcus thermophilus strain FBI3) were examined in different digestive compartments of gnotoxenic and human-microbiota-associated mice. The production of the lactose-hydrolysing enzyme (i.e. beta-galactosidase) was also investigated within the digestive tract, using a chromosomal reporter system based on luciferase genes from Photorhabdus luminescens under the control of the plac promoter. In both mice models, S. thermophilus cells transited within 2 h from the stomach to the caecum-colon compartment of the digestive tract where they displayed a survival rate of nearly 100 %. In gnotoxenic mice, luciferase activity was found to increase in the second half of the small intestine and in the caecum-colon compartment when lactose was added to the drinking water provided to the animals. In human-microbiota-associated mice drinking lactose, luciferase activity was similarly increased in the second half of the small intestine but was drastically reduced in the caecum-colon compartment. This feature could be ascribed to the presence of the resident human microbiota.

Animals↗

Comparison of a portable breath hydrogen analyser (Micro H2) with a Quintron MicroLyzer in measuring lactose maldigestion, and the evaluation of a Micro H2 for diagnosing hypolactasia.

The measurement of hydrogen in exhaled air and changes in the concentration of blood glucose and urine galactose excretion are indirect methods of diagnosing hypolactasia. The aim of this study was to compare a portable breath hydrogen analyser (Micro H2) with a widely used model (Quintron MicroLyzer) and to compare them with the blood glucose, urine galactose, and gastrointestinal symptoms in the lactose tolerance test. After an overnight fast, 44 volunteers (18-66 y) ingested 50 g lactose in a single oral dose. Changes in exhaled breath hydrogen concentrations were measured with the two analysers, and changes in blood glucose and urinary galactose were assayed for 4 h and used as a reference. Eighteen subjects were diagnosed as maldigesters according to our gold standard of at least two positive tests out of the three: breath hydrogen by Quintron, blood glucose concentration, and urine galactose excretion. The highest increase in the breath hydrogen concentration over the baseline was highly variable: 44-366 ppm (Micro H2) or 27-187 ppm (Quintron MicroLyzer). The sensitivity, specificity, and positive and negative predictive values of the Micro H2 compared to the gold standard were 83%, 96%, 94% and 89%, respectively. Overall agreement was 91% (95% CI 78-97%). Compared to the Quintron, the diagnoses were identical in 100% of the cases (92-100%). Thus, for diagnosing hypolactasia, the Micro H2 appeared as reliable for measuring breath hydrogen concentrations as Quintron MicroLyzer commonly used in oral lactose tolerance tests.

Adolescent↗

Viable starter culture, beta-galactosidase activity, and lactose in duodenum after yogurt ingestion in lactase-deficient humans.

Ten lactose malabsorbers were intubated and given fresh or heated yogurt to which polyethylene-glycol (PEG) and spores of Bacillus stearothermophilus (SBS) had been added as internal standards. In duodenal samples taken after fresh yogurt ingestion, viable starter culture was detected for 60 min in 6 of 7 subjects and the ratio of microbial beta-galactosidase activity to SBS remained similar during this period to its value in the preingested yogurt. In the two groups ingesting fresh and heated yogurt respectively, ratios of lactose to PEG remained similar to preingested values for 90 min and duodenal pH remained less than 5.1. In vitro, at pH 5.0, beta-galactosidase activity in yogurt dropped by 80%. These data clearly show that after fresh yogurt ingestion, viable starter culture reaches the duodenum and contains beta-galactosidase activity. However, the buffering capacity of the yogurt that protects bacteria from acidic gastric secretion also prevents microbial beta-galactosidase from hydrolyzing lactose in the duodenum.

Adult↗

Effect of lactic acid bacteria on the intestinal production of lactate and short-chain fatty acids, and the absorption of lactose.

The characteristic fermentation pattern seen with specific saccharides in incubations with pure cultures of Lactobacillus acidophilus and Bifidobacterium bifidum disappeared when incubated in 16.7% fecal homogenate. The productions of lactate and short-chain fatty acids in mixed bacterial-fecal incubations were similar to productions in fecal homogenates without L acidophilus and B bifidum and were mainly associated with the specific mono-, di-, and polysaccharides added to the incubate. B bifidum was cultured from ileostomic contents in eight of nine ileostomists after oral administration (2.4 x 10(10) cells), but did not influence the concentrations and productions of DL-lactate and short-chain fatty acids in the ileostomic outputs and incubates. Large amounts of ingested lactic acid bacteria (4.2 x 10(10) cells) did not ameliorate lactose malabsorption measured by the breath-hydrogen test in 12 lactose malabsorbers. This study shows that ingested lactic acid bacteria are indeed present in the colon, but it does not support the theory that they change the pattern of colonic fermentation or the degree of intestinal lactose malabsorption.

Adult↗

beta-Galactosidase from Aspergillus niger in adult lactose malabsorption: a double-blind crossover study.

An assessment was made of the efficacy of a beta-galactosidase, obtained from Aspergillus niger and added to intact milk, in decreasing lactose malabsorption and intolerance. Sixteen adult patients with malabsorption and intolerance to this sugar were studied in a double-blind crossover study vs. placebo. A 5-hour hydrogen breath test was used to assess malabsorption of lactose contained in 400 ml milk. When compared with placebo, the addition of exogenous lactase to intact milk caused a statistically significant reduction in the maximum breath H2 concentration (P less than 0.01) and in the cumulative H2 excretion (P less than 0.005). In the same way, the cumulative index for gastrointestinal intolerance was significantly lower (P less than 0.005) after the ingestion of lactase-added milk. This study demonstrates that enzyme replacement therapy, with beta-galactosidases obtained from Aspergillus niger, is effective in decreasing lactose malabsorption and its consequent intolerance in adult subjects with lactase deficiency.

Adult↗

Gut transit time and lactose malabsorption during phototherapy. II. A study using raw milk from the mothers of the infants.

Thirty newborn infants with normal birth weight suffering from uncomplicated hyperbilirubinemia were studied. 15 infants received ordinary phototherapy and 15 intensive phototherapy (blue double light). All infants received their mothers' milk or fresh milk from mothers of other newborn infants of the same age. All infants had normal lactose tolerance test during the phototherapy, except one infant receiving ordinary transit time was relatively long. The gut transit time was significantly shorter in the infants treated with intensive phototherapy than in those treated with ordinary phototherapy without there being any significant difference in the increase in blood glucose by lactose tolerance tests. It is concluded that lactose malabsorption is not the usual cause of the reduced gut transit time during phototherapy even if the infants receive their mothers' milk.

Bilirubin↗

Plasma insulin response to oral carbohydrate in patients with glucose and lactose malabsorption.

Plasma insulin levels were determined following oral glucose in 12 patients with adult coeliac disease, after oral lactose in four patients with alactasia, and in age-matched control subjects. In coeliac patients the insulin response was greater than expected from the small rise in blood sugar, and no correlation was found between plasma insulin and sugar levels at any period during the test. The separation of the plasma insulin curve from the blood sugar curve after glucose is in keeping with the concept that a factor responsible for stimulating insulin secretion is released from the gut during or after absorption of glucose. In patients with selective lactose malabsorption (alactasia) administration of lactose by mouth failed to elicit any insulin response, indicating that the insulin-releasing effect of the bowel is not activated merely by the presence of intraluminal carbohydrate.

Administration, Oral↗

[Genetic analysis of lactose-fermenting Salmonella typhimurium isolated in Rio de Janeiro].

Lactose fermenting Salmonella typhimurium are endemic in São Paulo, but not in Rio de Janeiro Two isolations are described from the latter city. These Rio de Janeiro strains have a plasmid of 7.4 megadaltons. These plasmids were not auto-transferable, were thermostable and were not eliminated by acridine orange. One of these strains arose from a plasmid that had the lactose operon repressed, leading us to speculate about the evolution of the lactose fermenting character in Brazilian Salmonella.

Brazil↗