Rye bran as fermentation matrix boosts in situ dextran production by Weissella confusa compared to wheat bran.

The consumption of fiber-rich foods such as cereal bran is highly recommended due to its beneficial health effects. Pre-fermentation of bran with lactic acid bacteria can be used to improve the otherwise impaired flavor and textural qualities of bran-rich products. These positive effects are attributed to enzymatic modification of bran components and the production of functional metabolites like organic acids and exopolysaccharides such as dextrans. The aim of this study was to investigate dextran production in wheat and rye bran by fermentation with two Weissella confusa strains. Bran raw materials were analyzed for their chemical compositions and mineral content. Microbial growth and acidification kinetics were determined from the fermentations. Both strains produced more dextran in rye bran in which the fermentation-induced acidification was slower and the acidification lag phase longer than in wheat bran. Higher dextran production in rye bran is expected to be due to the longer period of optimal pH for dextran synthesis during fermentation. The starch content of wheat bran was higher, which may promote isomaltooligosaccharide formation at the expense of dextran production. W. confusa Cab3 produced slightly higher amounts of dextran than W. confusa VTT E-90392 in all raw materials. Fermentation with W. confusa Cab3 also resulted in lower residual fructose content which has technological relevance. The results indicate that wheat and particularly rye bran are promising matrices for producing technologically significant amounts of dextran, which facilitates the use of nutritionally valuable raw bran in food applications.

Structure modeling and functional analysis of recombinant dextransucrase from Weissella confusa Cab3 expressed in Lactococcus lactis.

The dextransucrase gene from Weissella confusa Cab3, having an open reading frame of 4.2 kb coding for 1,402 amino acids, was amplified, cloned, and expressed in Lactococcus lactis. The recombinant dextransucrase, WcCab3-rDSR was expressed as extracellular enzyme in M17 medium with a specific activity of 1.5 U/mg which after purification by PEG-400 fractionation gave 6.1 U/mg resulting in 4-fold purification. WcCab3-rDSR was expressed as soluble and homogeneous protein of molecular mass, approximately, 180 kDa as analyzed by SDS-PAGE. It displayed maximum enzyme activity at 35°C at pH 5.0 in 50 mM sodium acetate buffer. WcCab3-rDSR gave Km of 6.2 mM and Vm of 6.3 µmol/min/mg. The characterization of dextran synthesized by WcCab3-rDSR by Fourier transform infrared and nuclear magnetic resonance spectroscopic analyses revealed the structural similarities with the dextran produced by the native dextransucrase. The modeled structure of WcCab3-rDSR using the crystal structures of dextransucrase from Lactobacillus reuteri (protein data bank, PDB id: 3HZ3) and Streptococcus mutans (PDB id: 3AIB) as templates depicted the presence of different domains such as A, B, C, IV, and V. The domains A and B are circularly permuted in nature having (ß/α)8 triose phosphate isomerase-barrel fold making the catalytic core of WcCab3-rDSR. The structure superposition and multiple sequence alignment analyses of WcCab3-rDSR with available structures of enzymes from family 70 GH suggested that the amino acid residue Asp510 acts as a nucleophile, Glu548 acts as a catalytic acid/base, whereas Asp621 acts as a transition-state stabilizer and these residues are found to be conserved within the family.

Luminescence nanothermometry using a trivalent lanthanide co-doped perovskite.

This study investigates in detail the laser-mediated upconversion emission and temperature-sensing capability of (Ca0.99-a Yb0.01Er a )TiO3. Samples were prepared at different concentrations to observe the effect of erbium on upconversion while increasing its concentration and keeping all the other parameters constant. Doping is a widespread technological process which involves incorporating an element called a dopant in a lower ratio to the host lattice to derive hybrid materials with desired properties. The (Ca0.99-a Yb0.01Er a )TiO3 perovskite nanoparticles were synthesized via a sol-gel technique. The frequency upconversion was performed using a 980 nm laser diode excitation source. X-ray diffractometry (XRD) confirmed that the synthesized samples are crystalline in nature and have an orthorhombic structure. The temperature-sensing ability was examined using the fluorescence intensity ratio (FIR) algorithm of two emission bands (2H11/2 → 4I15/2 and 4S3/2 → 4I15/2) of the Er3+ ion. Temperature-dependent upconversion luminescence is observed over a broad temperature range of 298-623 K. The maximum sensor sensitivity obtained is 6.71 × 10-3 K-1 at 110°.

CoQ10 a super-vitamin: review on application and biosynthesis.

Coenzyme Q10 (CoQ) or ubiquinone is found in the biological system which is synthesized by the conjugation of benzoquinone ring with isoprenoid chain of variable length. Coenzyme Q10 supplementation energizes the body and increases body energy production in the form of ATP and helps to treat various human diseases such as cardiomyopathy, muscular dystrophy, periodontal disease, etc. Reports of these potential therapeutic advantages of CoQ10 have resulted in its high market demand, which focus the researchers to work on this molecule and develop better bioprocess methods for commercial level production. At the moment, chemical synthesis, semi-synthetic method as well as bio-production utilizing microbes as biofactory are in use for the synthesis of CoQ10. Chemical synthesis involves use of cheap and easily available precursor molecules such as isoprenol, chloromethylquinone, vinylalane, and solanesol. Chemical synthesis methods due to the use of various solvents and chemicals are less feasible, which limits its application. The microbial production of CoQ10 has added advantages of being produced in optically pure form with high yield using inexpensive medium composition. Several bacteria, e.g., Agrobacterium, Paracoccus, Rhodobacterium, and yeast such as Candida, Rhodotorula are the potent ubiquinone producer. Some alternative biosynthetic pathway for designing of CoQ10 production coupled with metabolic engineering might help to increase CoQ10 production. The most common practiced strategy for strain development for commercial CoQ10 production is through natural isolation and chemical mutagenesis. Here, we have reviewed the chemical, semi-synthetic as well as microbial CoQ10 production in detail.

Synthesis, spectroscopic investigation, molecular docking and DFT studies of novel (2Z,4Z)-2,4-bis(4-chlorobenzylidene)-5-oxo-1-phenylpyrrolidine-3-carboxylic acid (BCOPCA).

The synthesized compound (2Z,4Z)-2,4-bis(4-chlorobenzylidene)-5-oxo-1-phenylpyrrolidine-3-carboxylic acid (BCOPCA) was characterised by Ultraviolet, FT-Infra Red, 1H, 13C Nuclear Magnetic Resonance and mass spectroscopy. The compound was further subjected to quantum chemical calculations at the level of density functional theory (DFT) using 6-31G (d,p) basis sets method with B3LYP and CAM-B3LYP hybrid functionals. The intramolecular interactions, polarizability, hyperpolarizability and nonlinear optical properties of the title compound were also incorporated in the study. The total first static hyperpolarizability (ß0 = 19.477 × 10-30 and 16.924 × 10-30 esu) value was also computed and indicated the title molecule as an interesting forthcoming NLO material. The other thermodynamic properties (entropy, heat capacity and zero vibrational energy) were also discussed. The study also includes NBO computations, complete vibrational assignments, Mulliken charges, UV-Visible spectral analysis and HOMO-LUMO energies. The regions of low and high electron density were obtained from MESP and ESP maps. The calculated parameters for BCOPCA using aforementioned functions are harmonious with the experimental findings. The in-vitro antimicrobial activity and molecular docking studies of BCOPCA were also done and showed a good correlation.

Optimization of Isomaltooligosaccharide Size Distribution by Acceptor Reaction of Weissella confusa Dextransucrase and Characterization of Novel α-(1→2)-Branched Isomaltooligosaccharides.

Long-chain isomaltooligosaccharides (IMOs) are promising prebiotics. IMOs were produced by a Weissella confusa dextransucrase via maltose acceptor reaction. The inputs of substrates (i.e., sucrose and maltose, 0.15-1 M) and dextransucrase (1-10 U/g sucrose) were used to control IMO yield and profile. According to response surface modeling, 1 M sucrose and 0.5 M maltose were optimal for the synthesis of longer IMOs, whereas the dextransucrase dosage showed no significant effect. In addition to the principal linear IMOs, a homologous series of minor IMOs were also produced from maltose. As identified by MS(n) and NMR spectroscopy, the minor trisaccharide contained an α-(1→2)-linked glucosyl residue on the reducing residue of maltose and thus was α-d-glucopyranosyl-(1→2)-[α-d-glucopyranosyl-(1→4)]-d-glucopyranose (centose). The higher members of the series were probably formed by the attachment of a single unit branch to linear IMOs. This is the first report of such α-(1→2)-branched IMOs produced from maltose by a dextransucrase.

Weissella confusa Cab3 dextransucrase: properties and in vitro synthesis of dextran and glucooligosaccharides.

Food-derived Weissella spp. have gained attention during recent years as efficient dextran producers. Weissella confusa Cab3 dextransucrase (WcCab3-DSR) was isolated applying PEG fractionation and used for in vitro synthesis of dextran and glucooligosaccharides. WcCab3-DSR had a molar mass of 178 kDa and was activated by Co(2+) and Ca(2+) ions. Glycerol and Tween 80 enhanced enzyme stability, and its half-life at 30°C increased from 10h to 74 h and 59 h, respectively. The (1)H and (13)C NMR spectral analysis of the produced dextran confirmed the presence of main chain α-(1→6) linkages with only 3.0% of α-(1→3) branching, of which some were elongated. An HPSEC analysis in DMSO revealed a high molecular weight of 1.8 × 10(7)g/mol. Glucooligosaccarides produced through the acceptor reaction with maltose, were analyzed with HPAEC-PAD and ESI-MS/MS. They were a homologous series of isomaltooligosaccharides with reducing end maltose units. To the best of our knowledge, this is a first report on native W. confusa dextransucrase.

16S rRNA-Based Identification of a Glucan-Hyperproducing Weissella confusa.

A gram-positive, nonmotile, irregular, short, rod-shaped new strain of Weissella confusa bacterium was isolated from fermented cabbage. The isolate was physiologically and biochemically characterised. The 16S rDNA was amplified by polymerase chain reaction (PCR). The isolate was identified as Weissella confusa (GenBank accession number: GU138518.1) based on nucleotide homology and phylogenetic analysis. The isolate produces glucansucrase when grown in sucrose-supplemented culture medium which catalyses glucan formation. This novel isolate possesses high capacity of industrial use due to its high productivity of glucan (34 mg/mL) as compared to other strains reported. The optimum temperature for glucansucrase production was 25°C. The shaking condition gave an enzyme activity of 6.1 U/mL which was 1.5 times higher than that given by static condition (4.1 U/mL). The temperature 35°C, pH 5.4, and ionic strength 10-20 mM were optimum for enzyme assay. This investigation unraveled the abundance of industrially valuable microflora of the north east India.

Optimization of Fermentation Medium for Enhanced Glucansucrase and Glucan Production from Weissella confusa

A glucan producing Weissella confusa isolated from fermented cabbage, as identified earlier, was used for optimization of its fermentation medium. The effects of various macronutrients such as sucrose, glucose as cosubstrate, yeast extract, beef extract, peptone, sodium acetate, K2HPO4 and Tween 80 were studied on glucansucrase and glucan production from Weissella confusa. The medium used as control gave 6.0 U/ml enzyme activity and 34 mg/ml glucan concentration. Sucrose (5 percent), glucose as cosubstrate (5 percent for glucansucrase and 3 percent for glucan production), Tween 80 (0.1 percent), yeast extract (1.5 percent), Peptone (2.0 percent) and K2HPO4 (1.5 percent) were effective nutrients displaying higher glucansucrase and glucan production giving 18.2, 18.0, 7.0, 6.4, 6.2 and 6.4 U/ml enzyme activity and 103, 100, 46, 41, 39 and 37 mg/ml glucan concentration, respectively. Sodium acetate and beef extract were not effective for enzyme and glucan production. The new strain Weissella confusa can be used for commercial production of glucansucrase and glucan using optimized medium.