Melibiose

About: Melibiose is a research topic. Over the lifetime, 1002 publications have been published within this topic receiving 27300 citations. The topic is also known as: Melibiose.

Gal4 Mutations That Separate the Transcriptional Activation and Gal80-Interactive Functions of the Yeast Gal4 Protein

Abstract: The carboxy-terminal 28 amino acids of the Saccharomyces cerevisiae transcriptional activator protein GAL4 execute two functions--transcriptional activation and interaction with the negative regulatory protein, GAL80. Here we demonstrate that these two functions are separable by single amino acid changes within this region. We determined the sequences of four GAL4C-mutations, and characterized the abilities of the encoded GAL4C proteins to activate transcription of the galactose/melibiose regulon in the presence of GAL80 and superrepressible GAL80S alleles. One of the GAL4C mutations can be compensated by a specific GAL80S mutation, resulting in a wild-type phenotype. These results support the idea that while the GAL4 activation function tolerates at least minor alterations in the GAL4 carboxyl terminus, the GAL80-interactive function is highly sequence-specific and sensitive even to single amino acid alterations. They also argue that the GAL80S mutations affect the affinity of GAL80 for GAL4, and not the ability of GAL80 to bind inducer.

Influence of Lipids with and without Other Cottonseed Reserve Materials on Aflatoxin B1 Production by Aspergillus flavus

Abstract: Cottonseed storage lipids (primarily triglycerides), in either crude or refined form, were found to support growth and aflatoxin B(1) production by Aspergillus flavus. When lipids were removed from ground whole cottonseed by petroleum ether extraction, aflatoxin production dropped by more than 800-fold. Reconstitution of the lipid-extracted ground whole seed with a crude preparation of cottonseed lipids restored aflatoxin production to the previous levels. Fungal utilization of the three major cottonseed reserve materials, raffinose, triglycerides (refined cottonseed oil), and cottonseed storage protein, was monitored in vitro over a 7 day fermentation period. The fermentation medium contained the reserve compounds in proportions approximating those found in mature cottonseed. A. flavus rapidly converted raffinose to fructose and melibiose, presumably by action of invertase, and then hydrolyzed the melibiose. These simple sugars apparently supported initial growth and aflatoxin B(1) production. Raffinose and the resulting melibiose were nearly exhausted by day 2. Fungal hydrolysis of triglycerides began as exhaustion of carbohydrate approached. After day 2, rapid catabolism of the released fatty acids began and coincided with glucose regeneration through gluconeogenesis, which peaked on day 6. The fungus did not preferentially utilize specific fatty acids. A. flavus also produced a number of storage metabolites, including arabitol, erythritol, mannitol, and trehalose. Mannitol was produced in much higher concentrations than the other storage metabolites. Selective use of simple carbohydrates by A. flavus to drive aflatoxin production may suggest strategies for reducing vulnerability of cottonseed to aflatoxin contamination.

Thermovenabulum ferriorganovorum gen. nov., sp. nov., a novel thermophilic, anaerobic, endospore-forming bacterium.

Abstract: A thermophilic, anaerobic, spore-forming bacterium (strain Z-9801T) was isolated from a terrestrial hydrothermal source in the Uzon caldera on the Kamchatka peninsula. Cells of strain Z-9801T were straight, sometimes branched rods, 0.5-0.6 microm in diameter and 1.5-7.0 microm in length, with peritrichous flagella. The temperature range for growth was 45-76 degrees C, with an optimum at 63-65 degrees C. The pH range for growth was 4.8-8.2, with an optimum at 6.7-6.9. The substrates utilized by strain Z-9801T included peptone, yeast extract, beef extract, Casamino acids, starch, pyruvate, melibiose, sucrose, fructose, maltose, xylose and ribose. The fermentation products from melibiose were ethanol, acetate, H2 and CO2. Strain Z-9801T used H2 in the presence of Fe(III) and an organic electron donor. Strain Z-9801T reduced Fe(III), Mn(IV), nitrate, fumarate, sulfite, thiosulfate, elemental sulfur and 9,10-anthraquinone 2,6-disulfonate. The G+C content of strain Z-9801T DNA was 36 mol%. 16S rDNA sequence analysis revealed that the isolated organism forms a separate branch within the Bacillus/Clostridium group. On the basis of physiological properties and phylogenetic analysis, it is proposed that strain Z-9801T (= DSM 14006T = UNIQEM 210T) should be assigned to a novel species of a new genus, Thermovenabulum ferriorganovorum gen. nov., sp. nov.

Characterization of Genes Involved in the Metabolism of α-Galactosides by Lactococcus raffinolactis

Abstract: Lactococcus raffinolactis, unlike most lactococci, is able to ferment alpha-galactosides, such as melibiose and raffinose. More than 12 kb of chromosomal DNA from L. raffinolactis ATCC 43920 was sequenced, including the alpha-galactosidase gene and genes involved in the Leloir pathway of galactose metabolism. These genes are organized into an operon containing aga (alpha-galactosidase), galK (galactokinase), and galT (galactose 1-phosphate uridylyltransferase). Northern blotting experiments revealed that this operon was induced by galactosides, such as lactose, melibiose, raffinose, and, to a lesser extent, galactose. Similarly, alpha-galactosidase activity was higher in lactose-, melibiose-, and raffinose-grown cells than in galactose-grown cells. No alpha-galactosidase activity was detected in glucose-grown cells. The expression of the aga-galKT operon was modulated by a regulator encoded by the upstream gene galR. The product of galR belongs to the LacI/GalR family of transcriptional regulators. In L. lactis, L. raffinolactis GalR acted as a repressor of aga and lowered the enzyme activity by more than 20-fold. We suggest that the expression of the aga operon in lactococci is negatively controlled by GalR and induced by a metabolite derived from the metabolism of galactosides.

Purification and characterization of the raffinose oligosaccharide chain elongation enzyme, galactan: galactan galactosyltransferase (GGT), from Ajuga reptans leaves

Abstract: Galactan: galactan galactosyltransferase (GGT), an enzyme involved in the biosynthesis of the long-chain raffinose family of oligosaccharides (RFOs) in Ajuga reptans, catalyses the transfer of an alpha-galactosyl residue from one molecule of RFO to another one resulting in the next higher RFO oligomer This novel galactinol (alpha-galactosyl-myo-inositol)-independent alpha-galactosyltransferase is responsible for the accumulation of long-chain RFOs in vivo Warm treatment (20 degrees C) of excised leaves resulted in a 34-fold increase of RFO concentration and a 200-fold increase of GGT activity after 28 days Cold treatment (10 degrees C/3 degrees C day/night) resulted in a 26- and 130-fold increase, respectively These data support the role of GGT as a key enzyme in the synthesis and accumulation of long-chain RFOs GGT was purified from leaves in a 4-step procedure which involved fractionated precipitation with ammonium sulphate as well as lectin affinity, anion exchange, and size-exclusion chromatography and resulted in a 200-fold purification Purified GGT had an isoelectric point of 47, a pH optimum around 5, and its transferase reaction displayed saturable concentration dependence for both raffinose (Km = 42 mM) and stachyose (Km = 58 mM) GGT is a glycoprotein with a 10% glycan portion The native molecular mass was 212 kDa as determined by size-exclusion chromatography Purified GGT showed one single active band after native PAGE or IEF separation, respectively, which separated into three bands on SDS-PAGE at 48 kDa, 66 kDa, and 60 kDa The amino acid sequence of four tryptic peptides obtained from the major 48-kDa band showed a high homology to plant alpha-galactosidase (EC 32122) sequences GGT differed, however, in its substrate specificity from alpha-galactosidases; it neither hydrolysed nor transferred alpha-galactosyl-groups from melibiose, galactinol, UDP-galactose, manninotriose, and manninotetrose Galactinol, sucrose, and galactose inhibited the GGT reaction considerably at 10-50 mM