ABSTRACT
The draft genome sequence of Lactobacillus plantarum Kanjika 2007, isolated from the South Indian staple, medicinal, and traditional food kanjika, is reported here. The whole genome consists of 3.16 Mb with a G+C content of 44.7% and 3,009 protein-coding genes, 78 tRNAs, and 4rRNAs (5S-23S-16S).
ABSTRACT
Accumulating evidence suggests that probiotic bacteria play a vital role in modulating various aspects integral to the health and well-being of humans. In the present study, probiotic attributes and the antioxidant, anti-inflammatory and neuromodulatory potential of Enterococcus faecium CFR 3003 were investigated by employing suitable model systems. E. faecium exhibited robust resistance to gastrointestinal stress conditions as it could withstand acid stress at pH 1.5, 2 and 3. The bacterium also survived at a bile salt concentration of 0.45 %, and better tolerance was observed towards pepsin and trypsin. E. faecium produced lactic acid as a major metabolic product, followed by butyric acid. Lyophilized cell-free supernatant (LCS) of E. faecium exhibited significant antioxidant capacity evaluated against 1,1-diphenyl-2-picryl-hydrazyl, ascorbate auto-oxidation, oxygen radical absorbance and reducing power. Interestingly, E. faecium, Lactobacillus rhamnosus GG MTCC 1408 and LCS showed a significant anti-inflammatory effect by negatively modulating TNF-α production and upregulating IL-10 levels in LPS-stimulated macrophage cell lines. In an in vivo mice model, the propensity of probiotic supplements to modulate endogenous oxidative markers and redox status in brain regions was assessed. Young mice provided with oral supplements (daily for 28âdays) of E. faecium and L. rhamnosus exhibited diminished oxidative markers in the brain and enhanced activities of antioxidant enzymes with a concomitant increase in γ-aminobutyric acid and dopamine levels. Collectively, our findings clearly suggest the propensity of these bacteria to protect against tissue damage mediated through free radicals and inflammatory cytokines. Although the underlying molecular mechanisms need further studies, it is tempting to speculate that probiotics confer a neuroprotective advantage in vivo against oxidative damage-mediated neurodegenerative conditions.
Subject(s)
Enterococcus faecium/physiology , Probiotics , Animals , Anti-Bacterial Agents/pharmacology , Antioxidants/metabolism , Antioxidants/pharmacology , Anxiety , Behavior, Animal , Cecum/microbiology , Cell Line , Enterococcus faecium/drug effects , Hydrogen-Ion Concentration , Interleukin-10/genetics , Interleukin-10/metabolism , Lacticaseibacillus rhamnosus , Macrophages/metabolism , Macrophages/microbiology , Male , Mice , Oxidative Stress , Pepsin A/pharmacology , Protein Carbonylation , Random Allocation , Time Factors , Trypsin/pharmacology , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Prebiotic oligosaccharides are demonstrated to confer a wide spectrum of physiological benefits during pregnancy. In view of this, focused attempts are being directed towards understanding their role as modulators of brain chemistry and behavior. Epidemiological studies have identified that exposure to neurotoxins during prenatal/early life can profoundly impact neurodevelopment/function. In this context, we have tested the hypothesis that a combination of prebiotic supplements during gestation has the propensity to attenuate acrylamide (ACR) induced oxidative impairments, mitochondrial dysfunction and neurotoxicity in maternal and fetal brain of rats. To achieve this, pregnant dams given oral supplements of a combination of fructo- and xylooligosaccharides (FOS + XOS, 3 g/kg/day) during gestation days (GD 0-19) were exposed to ACR (200 ppm in drinking water, GD 6-19). The behavioral analysis revealed that ACR dams fed prebiotics displayed higher exploratory behavior in the open field test. The prenatal evaluation showed that ACR-induced decrements of placental/fetal weights were markedly restored with prebiotic feeding. Prebiotics significantly offset markers of oxidative stress, restored enzymic antioxidants, cholinergic and mitochondrial function in the maternal and fetal brain. Concomitantly, prebiotics restored ACR-induced depletion in the levels of dopamine and γ-aminobutyric acid in the maternal cortex that positively correlated with cecal bacterial numbers. Collectively, these data suggest that prenatal prebiotic oligosaccharide supplements protect developing brain against oxidative stress-mediated neurotoxicity. While the underlying mechanism/s by which prebiotics abrogate the impact of neurotoxicants in the developing brain merits further studies, we speculate that it may be mediated predominantly through attenuation of oxidative stress and proliferation of enteric microbiota.
Subject(s)
Acrylamide/toxicity , Fructose/chemistry , Maternal Exposure , Maternal-Fetal Exchange/drug effects , Oligosaccharides/pharmacology , Oxidative Stress/drug effects , Xylose/chemistry , Animals , Cecum , Feeding Behavior/drug effects , Female , Male , Oligosaccharides/chemistry , Prebiotics , Pregnancy , Rats , Rats, Wistar , Weight Gain/drug effectsABSTRACT
Fructooligosaccharides (FOSs) were prepared from sucrose using fungal fructosyl transferase (FTase) obtained from Aspergillus oryzae MTCC 5154. The resulting mixture consisted of glucose (28-30%), sucrose (18-20%) and fructooligosaccharides (50-54%) as indicated by HPLC analysis. Identification of oligomers present in the mixture of fructooligosaccharides was carried out using NMR spectroscopy and LC-MS. No compounds other than mono-, di-, tri-, tetra- and pentasaccharides were identified in the FOS mixture prepared using FTase. NMR and LC-MS spectra proved the absence of any toxic microbial metabolites of Aspergillus species in FOS thereby emphasizing its safe use as a food ingredient. Animal studies conducted on streptozotocin-induced diabetic rats suggested that the use of FOS as an alternative non-nutrient sweetener is without any adverse effects on various diabetes-related metabolic parameters. Despite the high free-sugar content associated with it, FOS did not further aggravate the hyperglycemia and glucosuria in diabetic animals, even at 10% levels. On the other hand, by virtue of its soluble fibre effect, it has even alleviated diabetic-related metabolic complications to a certain degree.
Subject(s)
Fructose/chemistry , Oligosaccharides/chemistry , Sweetening Agents/chemistry , Animals , Aspergillus/enzymology , Diabetes Mellitus , Food Additives/chemistry , Hexosyltransferases/metabolism , Hyperglycemia/diet therapy , Magnetic Resonance Spectroscopy , Mass Spectrometry , Oligosaccharides/administration & dosage , Rats , Sucrose/metabolism , Sweetening Agents/administration & dosageABSTRACT
Fructosyl transferase (FTase) production by Aspergillus oryzae CFR 202 was carried out by solid-state fermentation (SSF), using various agricultural by-products like cereal bran, corn products, sugarcane bagasse,cassava bagasse (tippi) and by-products of coffee and tea processing. The FTase produced was used for the production of fructo-oligosaccharides (FOS), using 60% sucrose as substrate. Among the cereal bran used, rice bran and wheat bran were good substrates for FTase production by A. oryzae CFR 202. Among the various corn products used, corn germ supported maximum FTase production, whereas among the by-products of coffee and tea processing used, spent coffee and spent tea were good substrates, with supplementation of yeast extract and complete synthetic media. FTase had maximum activity at 60 degrees C and pH 6.0. FTase was stable up to 40 degrees C and in the pH range 5.0-7.0. Maximum FOS production was obtained with FTase after 8 h of reaction with 60% sucrose. FTase produced by SSF using wheat bran was purified 107-fold by ammonium sulphate precipitation (30-80%), DEAE cellulose chromatography and Sephadex G-200 chromatography. The molecular mass of the purified FTase was 116.3 kDa by SDS-PAGE. This study indicates the potential for the use of agricultural by-products for the efficient production of FTase enzyme by A. oryzae CFR 202 in SSF, thereby resulting in value addition of those by-products.
Subject(s)
Aspergillus oryzae/metabolism , Hexosyltransferases/biosynthesis , Hexosyltransferases/isolation & purification , Ammonium Sulfate/chemistry , Cellulose/metabolism , Chemical Fractionation , Chromatography, Gel , Chromatography, Ion Exchange , Coffee/metabolism , Culture Media/chemistry , Dietary Fiber/metabolism , Electrophoresis, Polyacrylamide Gel , Enzyme Stability , Fermentation , Hexosyltransferases/metabolism , Hydrogen-Ion Concentration , Molecular Weight , Oligosaccharides/metabolism , Peptones/metabolism , Sucrose/metabolism , Tea/metabolism , Temperature , Time Factors , Zea mays/metabolismABSTRACT
Immobilized lipase from Rhizomucor miehei (Lipozyme IM-20) was used to catalyze the esterification reaction between isovaleric acid and ethanol to synthesize ethyl isovalerate in n-hexane. Response surface methodology based on five-level four-variable central composite rotatable design was employed to optimize four important reaction variables such as enzyme/substrate E/S ratio, substrate concentration, incubation time, and temperature affecting the synthesis of ethyl isovalerate. The optimum conditions predicted for achieving maximum ester yield (500 mM) are as follows: E/S ratio, 48.41 g/mol; substrate concentration, 1 M; reaction time, 60 h; temperature, 60 degrees C. The predicted value matched well with experimentally obtained value of 487 mM.
Subject(s)
Lipase/chemistry , Rhizomucor/chemistry , Valerates/chemical synthesis , Analysis of Variance , Catalysis , Enzymes, Immobilized , Esterification , Ethanol/chemistry , Hemiterpenes , Hexanes , Mathematical Computing , Pentanoic Acids/chemistry , Substrate Specificity , Temperature , Time FactorsABSTRACT
The effects of important reaction parameters for enhancing isoamyl acetate formation through lipase-catalyzed esterification of isoamyl alcohol were investigated in this study. Increase in substrate (acid) concentration led to decrease in conversions. A critical enzyme concentration of 3 g l(-1) was detected for a substrate concentration of 0.06 M (each of alcohol and acid). Solvents with partition coefficient higher than 1000 (log P>3.0) supported enzyme activity to give high conversions. Acetic acid at higher concentrations could not be esterified easily probably owing to its role in lowering the microaqueous pH of the enzyme. Extraneous water/buffer addition decreased the isoamyl acetate yields slightly ( approximately 10%) at 0.005-0.01% v/v of the reaction mixture and drastically (>40%) at above 0.01% v/v. Buffer saturation of the organic solvent employed improved esterification (upto two-fold), particularly at moderately higher substrate concentrations (>0.18 M). Employing acetic anhydride instead of acetic acid resulted in a two-fold increase in the yields (at 0.25 M substrate). Use of excess nucleophile (alcohol) concentration by increasing the alcohol/acid molar ratio resulted in higher conversions in shorter duration (upto eight-fold even at 1.5 M acetic acid). Yields above 80% were achieved with substrate concentrations as high as 1.5 M and more than 150 g l(-1) isoamyl acetate concentrations were obtained employing a relatively low enzyme concentration of 10 g l(-1). The operational stability of lipase was also observed to be reasonably high enabling ten reuses of the biocatalyst.
Subject(s)
Enzymes, Immobilized/metabolism , Lipase/metabolism , Pentanols/chemical synthesis , Rhizomucor/enzymology , Biotechnology/methods , Buffers , Enzyme Stability , Enzymes, Immobilized/chemistry , Lipase/chemistry , Pentanols/metabolism , SolventsABSTRACT
The lactone 6-pentyl-alpha-pyrone has a characteristic coconut aroma and is produced by Trichoderma species. A study on the fermentative production of 6-pentyl-alpha-pyrone in both surface and submerged conditions by Trichoderma harzianum was carried out. Maximum concentrations of 455 mg/l and 167 mg/l after 96 h and 48 h of fermentation in surface and submerged conditions, respectively, were obtained without using any additional recovery operations. The resultant yields are higher than those previously reported in the literature, which may be attributable to strain characteristics in combination with the choice of fermentation conditions employed in the present study. Enough scope exists for further improvement in the yields by optimizing the cultural and nutritional parameters.
Subject(s)
Pyrones/metabolism , Trichoderma/metabolism , Biomass , Culture Media , Fermentation , Lactones/metabolism , Trichoderma/growth & developmentABSTRACT
In this study, an attempt has been made to maximize the lipid production using response surface methodology (RMS). Levels of nitrogen, carbon, and inoculum were chosen as factors. The results indicated that inoculum level was very important in lipid production, followed by carbon and nitrogen levels. At higher levels of inoculum, the strain was found to be more tolerant to higher concentrations of sugar, and significantly increased lipid production was noticed. Through the fitted models of second order, as per RSM, carbon at 10.24%, nitrogen at 0.37 g/L, and inoculum at the 20% level resulted in maximum biosynthesis of lipids.
ABSTRACT
Comprehensive studies of pure colonies of Streptomyces fradiae in the production of xylose isomerase by submerged fermentation at shake flask level revealed poor culture stability with respect to enzyme production, biomass formation, degree of pigmentation, quantity of glucose and xylose utilization, level of enzyme in cell-free culture broth and final pH of the fermentation medium. The results serve to stress obligatory evaluation of culture stability of Streptomyces strains in determining their suitability for use in developing fermentation processes for commercial exploitation.