Parvinder Kaur

Yeast Phytases: Present Scenario and Future Perspectives

ABSTRACT Phytases hydrolyze phytates to liberate soluble and thus readily utilizable inorganic phosphate. Although phytases are produced by various groups of microbes, yeasts being simple eukaryotes and mostly non-pathogenic with proven probiotic benefits can serve as ideal candidates for phytase research. The full potential of yeast phytases has not, however, been exploited. This review focuses attention on the present status of knowledge on the production, characterization, molecular characteristics, and cloning and over-expression of yeast phytases. Several potential applications of the yeast phytases in feeds and foods, and in the synthesis of lower myo-inositol phosphates are also discussed.

Effect of 1-Oxo-5β, 6β-Epoxy-Witha-2-Ene-27-Ethoxy-Olide Isolated from the Roots of Withania somnifera on Stress Indices in Wistar Rats

OBJECTIVE Isolation of biologically active fractions and compounds from the roots of Withania somnifera, a plant used extensively as a constituent of rasayana, in Ayurveda and to test their adaptogenic activity on stress indices using the cold-hypoxia-restraint (C-H-R) model. DESIGN Bioactivity-guided fractionation of an aqueous extract of the roots of Withania somnifera led to the isolation of a new species of withanolide 1-oxo-5beta, 6beta-epoxy-witha-2-ene-27-ethoxy-olide. Structure elucidation, was carried out using proton nuclear magnetic resonance, infrared (IR), ultraviolet (UV), and mass spectroscopic analysis. Stress-related indices were evaluated, namely serum creatine phosphokinase (CPK) activity, serum lactate dehydrogenase (LDH) activity, serum corticosterone levels, and serum lipid peroxidation (LPO) levels. RESULTS There was a significant decrease in a serum CPK, LDH, and LPO levels in animals pretreated with (1) fraction-I (20 mg/kg body weight), (2) 1-oxo-5beta, 6beta-epoxy-witha-2-ene-27-ethoxy-olide (2.5 mg/kg body weight) in comparison to control when subjected to C-H-R stress. CONCLUSIONS The results show that the a new species of withanolide, 1-oxo-5beta, 6beta-epoxy-witha-2-ene-27-ethoxy-olide (compound-1) could prove to be an effective agent to counteract C-H-R stress.

Pphy - a cell-bound phytase from the yeast Pichia anomala: molecular cloning of the gene PPHY and characterization of the recombinant enzyme.

The Pichia anomala gene PPHY, which codes for a cell-bound phytase, was isolated from genomic DNA by PCR, using oligonucleotide sequences derived from the N-terminal region of the purified phytase protein (Pphyp) and a degenerate primer derived from conserved sequences of yeast and fungal phytases as primers. The gene harbours an ORF of 1389bp, encoding a 462-amino-acid protein. The deduced amino acid sequence has similarity, to a varied extent, with those of phosphatases from Pichia stipitis (62%), Candida dubliniensis (51%), Candida albicans (51%), Arxula adeninivorans (35%) and phytases from Debaryomyces castellii (50%) and Pichia fabianii (39%). The sequence contains the phytase consensus heptapeptide motif (-Arg-His-Gly-X-Arg-X-Pro-) as well as two phosphohistidine signature motifs found in histidine acid phosphatases. After transformation of PPHY into the yeasts Saccharomyces cerevisiae, A. adeninivorans and Hansenula polymorpha, the last species was selected as the most suitable for synthesis of recombinant Pphyp. The cell-bound enzyme activities produced by wild-type P. anomala and transgenic H. polymorpha strains bearing the PPHY gene placed under the control of the inducible H. polymorpha-derived FMD promoter were characterized. In both cases, a molecular mass of approximately 380kDa was determined for the native enzyme (corresponding to a hexamer); the pH and temperature optima for the activity were 4.0 and 60 degrees C, respectively. The enzyme was active on phytic acid, p-nitrophenylphosphate, glucose-6-phosphate, ADP, sodium pyrophosphate, AMP, 1-naphthylphosphate and ATP. Based on the K(m)/K(cat) and further biochemical parameters, the enzyme was classified as a cell-bound phytase with acid phosphatase activity and not as acid phosphatase, despite its strong similarity to the latter class of enzymes. The yeast biomass containing phytase has been demonstrated to be useful as a feed additive in poultry and aquaculture, and dephytinization of foods and feeds.

Metalloporphyrins—Applications and clinical significance

The fascinating structures of naturally occurring porphyrins and metalloporphyrins have been perfected by nature to give functional dyes par excellence. The important roles these tetrapyrrolic macrocycles play in vital biological processes, in particular photosynthesis (chlorophyll), oxygen transport (hemoglobin), oxygen activation (cytochrome), have led to their characterization as ‘pigments of life’. Because porphyrins possess extended π-electron systems and exhibit stability, they are finding use, to an increasing extent, in advanced materials, as components in organic metals, molecular wires, and other devices. In medicine, porphyrins are experiencing a renaissance due to the advent of photodynamic therapy of great promise in the treatment of cancer and dermatological diseases. The interdisciplinary interest porphyrins thus generate has provided the impetus to develop Novel-porphyrin like molecules anticipated to exhibit special properties, by structural variation of the tetrapyrrolic macrocycle, while maintaining a (4n+2)π main conjugation pathway.In addition to their esoteric application in science, porphyrins have been shown to have profound implications for therapeutic purposes. Their photosensitizing properties have led to their utilization in photodynamic therapy. Certain metalloporphyrins such as SnPP are being tested as drugs for the treatment of neonatal jaundice. Metalloporphyrins are serving as SOD mimetics to combat oxidative stress and a range of metalloporphyrin complexes have been proposed as contrast agents for magnetic resonance imaging

Production, characteristics and applications of the cell-bound phytase of Pichia anomala.

Among several yeasts isolated from dried flowers of Woodfordia fruticosa, Pichia anomala produced a high titre of cell-bound phytase. The optimization of fermentation variables led to formulation of media and selection of cultural variables that supported enhanced phytase production. The enzyme productivity was very high in fed batch fermentation in air-lift fermentor as compared to that in stirred tank fermentor. Amelioration in the cell-bound phytase activity was observed when yeast cells were permeabilized with Triton-X-100. The enzyme is thermostable and acid stable with broad substrate specificity, the characteristics that are desirable for enzymes to be used in the animal feed industry. The phytase-encoding gene was cloned and sequenced. The 3D structure of the enzyme was proposed by comparative modeling using phytase of Debaryomyces occidentalis (50% sequence identity) as template. When broiler chicks, and fresh water and marine fishes were fed with the feed supplemented with yeast biomass containing phytase, improvement in growth and phosphorus retention, and decrease in the excretion of phosphorus in the faeces were recorded. The cell-bound phytase of P. anomala could effectively dephytinize wheat flour and soymilk.

A biologically active constituent of withania somnifera (ashwagandha) with antistress activity.

In Ayurvedic medicine,Withania somnifera (Ashwagandha) is well known for its anti-stress activity. A passive rat experimental model, where the animals are subjected to multiple stress of cold, hypoxia, restraint (C-H-R) has been developed to evaluate adaptogenic properties of various fractions of W. somnifera root extracts. We have carried out extraction of roots of W. somnifera with water and further isolated one of the active constituents called compound X and also tested its anti-stress activity in C-H-R model. The effect of administration of W. somnifera water suspension (360 mg/Kg bw) and compound X (20mg/Kg bw) on the fall and recovery of colonic temperature was noted. There was an increase of ≈38% and ≈54% in the time taken to attain Trec 23°C by rats given a single dose of fresh aqueous suspension and biologically active constituent (Compound X) respectively, where as decrease in the recovery time to attain Trec 37°C is ≈13% and ≈33% respectively, as compared to control group. It is clear that rats treated with the fresh aqueous suspension and compound X ofWithania somnifera, could withstand the multiple stress of C-H-R better than control group.

Phase integral corrections to radio wave absorption and virtual height for model ionospheric layers

Abstract The phase integral solution of the wave equation is a fairly good approximation in computing the radio wave absorption and virtual height for a slowly varying ionosphere. In the present paper we have used different ionospheric electron density models for E- and F-region heights and constant electron collision frequency for calculating the phase integral corrections to absorption and virtual height. It is found that the maximum corrections to absorption and virtual height for O-mode are of the order of 6 per cent and 3 per cent respectively, whereas for the X-mode they are comparatively smaller. It is also found that with increasing geomagnetic dip angle the corrections for the O-mode increase while those for the X-mode decrease. The results for the O-mode are in good agreement with the approximate expressions for absorption and virtual height given by earlier workers. However, no such explicit relations are available for the X-mode.