Vinod Chhokar

Effect of Additives on the Activity of Tannase from Aspergillus awamori MTCC9299

Tannase from Aspergillus awamori MTCC 9299 was purified using ammonium sulfate precipitation followed by ion-exchange chromatography. A purification fold of 19.5 with 13.5% yield was obtained. Temperature of 30 °C and pH of 5.5 were found optimum for tannase activity. The effects of metals and organic solvents on the activity of tannase were also studied. Metal ions Mg+2, Mn+2, Ca+2, Na+, and K+ stimulated the tannase activity, while Cu+2, Fe+3, and Co+2 acted as inhibitors of the enzyme. The addition of organic solvents like acetic acid, isoamylalcohol, chloroform, isopropyl alcohol, and ethanol completely inhibited the enzyme activity. However, butanol and benzene increased the enzyme activity.

Recent Advances in Industrial Application of Tannases: A Review

Tannin acyl hydrolase (E.C. 3.1.1.20) commonly referred as tannase, is a hydrolytic enzyme that catalyses the hydrolysis of ester bonds present in gallotannins, ellagitannins, complex tannins and gallic acid esters. Tannases are the important group of botechnologically relevant enzymes distributed throughout the animal, plant and microbial kingdoms. However, microbial tannases are currently receiving a great deal of attention. Tannases are extensively used in food, feed, pharmaceutical, beverage, brewing and chemical industries. Owing to its diverse area of applications, a number of patents have been appeared in the recent past. The present review pretends to present the advances and perspectives in the industrial application of tannase with special emphasis on patents.

Effect of garlic (Allium sativum L.) extract on degree of hydration, fructose, sulphur and phosphorus contents of rat eyelens and intestinal absorption of nutrients.

Influence of aqueous garlic extract on degree of hydration, fructose, sulphur and phosphorus contents of rat eyelens and intestinal absorption of nutrients were assessed. Inclusion of garlic extract in culture medium containing glucose and xylose inhibited the hydration of rat eyelens, whereas galactose evinced the reverse trend. Aqueous garlic extract in general decreased the concentration of fructose and phosphorus, whereassulphur concentration increased when rat eyelenses, were incubated with galactose and xylose. Garlic extract inhibited intestinal absorption of glutamic acid, sucrose and glucose to different extents. The rate of absorption of glutamic acid was found to be considerably higher than that of glucose and sucrose.

Recent Advances in Phytoremediation Technology

The rapid increase in the environmental contaminants due to various anthropogenic activities has become a serious issue worldwide. New and efficient measures are explored to remove or contain the threat from the increasing levels of environmental pollution. Plant-based soil and water remediation (phytoremediation) is one such method which can prove to be a sustainable and promising treatment to remediate environmental problems. Phytoremediation exploits the abilities of green plants to uptake, stabilize, or metabolize the pollutants. Moreover, it is a cost-effective and environmentally safe approach as compared to conventional methods to solve the problems of soil and water pollution. Phytoremediation technique has been successfully applied to treat various contaminated sites and pollutants such as heavy metals, dyes, fly ash, hydrocarbons etc. and furthermore, research is underway for exploring new ways to improve the phytoremediation process.

Bioremediation of Tannery Wastewater

Tannery effluent is a serious environmental threat due to its high chemical levels which include salinity, organic load (chemical oxygen load or demand, biological oxygen demand), inorganic matter, dissolved and suspended solids, ammonia, total Kjeldahl nitrogen, sulfide, chromium, chloride, sodium and other salt residues, heavy metals, etc. These components present in the effluent affect agriculture, human beings and livestock. Exposure to chromium and other pollutants in tannery effluent increases the risk of dermatitis, ulcer, nasal septum perforation and lung cancer. The environmental protection regulations stipulate that industries are not allowed to emit sulfide and chromium in the wastewater. Thus, removal of these high-strength toxic chemicals from the wastewater is very important. Treatment of tannery wastewater is carried out by physical, chemical, biological, or combination of these methods. Biological treatment of wastewater is more favorable and cost effective as compared to other physiochemical methods. A number of bioremediation strategies have been reported in the recent past showing their potential in the treatment of tannery effluent. The present review summarizes the recent advances in bioremediation of tannery effluent.

Molecular differentiation of Peroxysome proliferator activated receptor coactivator-1 among different breeds of Bubalus bubalis

Peroxysome proliferator activated receptor coactivator-1 gene (PPARGC1A) is a positional and functional candidate gene for milk fat yield. It has key role in energy, fat and glucose metabolism. Single nucleotide polymorphisms (SNPs) in Exon-8 of PPARGC1A are reported to be associated with milk fat yield in dairy cattle. In the present investigation PPARGC1A was partially amplified (around 767bp) by designing gene specific primer and confirm by sequencing the amplicon and its comparison with the PPARGC1A gene of bovine. Comparative study of PPARGC1A among different breeds of buffaloes reveals different level of mutations with respect to its gene sequence 0.013-1.69% and protein sequence 0.42% to 2.99%, Similarly the protein structures modeled from their sequences were compared by structural superposition that shows variations (RMSD) from 0.736 to 1.507. Furthermore, the sequences were used to generate a dendrogram. It reveals that Murrah and reference are very close to each other, similarly Toda, Bhadawari and Surti are closely related, whereas Pandharpuri is separated from both the cluster. Especially the variations are more at the binding site of this protein that may be the cause that different breeds have different percentage of milk fat. Further study is underway to detect polymorphism and associate them with milk fat related traits in buffalo.

Molecular Structure, Biological Functions, and Metabolic Regulation of Flavonoids

Flavonoids, constituting about 5–10% of the known secondary metabolites, are important for plant growth and development and thus can have a large impact on agricultural productivity. Plants are specialized in synthesizing and accumulating specific combinations of about 5000 flavonoids. The richness of flavonoids in plant sources cannot be underestimated and they are found closely associated with human daily dietary intake; hence, it is significant to understand the biochemistry of flavonoids. The flavonoid biosynthetic pathway is under tight developmental control, and multiple environmental factors like temperature lights stress and fertilizers affect the expression of flavonoid biosynthetic genes. Flavonoids exhibit a wide range of biological effects such as antibacterial, antiviral, antifungal, antiosteoporotic, antitumor, antithrombogenic, anti-inflammation, antiatherosclerotic, and antihepatotoxic activities. They also have a regulatory role on different hormones like estrogens, androgens, and thyroid hormones. Due to their diverse pharmacological activities, functional artificial polymeric flavonoids, flavonoid polymers, and amine-containing polymer-flavonoid conjugates have also been developed. As research advances in natural products, further achievements will certainly lead to a new age of flavonoids as either food supplements or pharmaceuticals.

Analytical profiling of mutations in quinolone resistance determining region of gyrA gene among UPEC

Mutations in gyrA are the primary cause of quinolone resistance encountered in gram-negative clinical isolates. The prospect of this work was to analyze the role of gyrA mutations in eliciting high quinolone resistance in uropathogenic E.coli (UPEC) through molecular docking studies. Quinolone susceptibility testing of 18 E.coli strains isolated from UTI patients revealed unusually high resistance level to all the quinolones used; especially norfloxacin and ciprofloxacin. The QRDR of gyrA was amplified and sequenced. Mutations identified in gyrA of E.coli included Ser83Leu, Asp87Asn and Ala93Gly/Glu. Contrasting previous reports, we found Ser83Leu substitution in sensitive strains. Strains with S83L, D87N and A93E (A15 and A26) demonstrated norfloxacin MICs ≥1024mg/L which could be proof that Asp87Asn is necessary for resistance phenotype. Resistance to levofloxacin was comparatively lower in all the isolates. Docking of 4 quinolones (ciprofloxacin, ofloxacin, levofloxacin and norfloxacin) to normal and mutated E.coli gyrase A protein demonstrated lower binding energies for the latter, with significant displacement of norfloxacin in the mutated GyrA complex and least displacement in case of levofloxacin.

De novo sequencing, assembly and characterisation of Aloe vera transcriptome and analysis of expression profiles of genes related to saponin and anthraquinone metabolism

BackgroundAloe vera is a perennial, succulent, drought-resistant plant that exhibits many pharmacological characteristics such as wound healing ability against skin burns, anti-ulcer, anti-inflammatory, anti-tumor, anti-viral, anti-hypercholesterolemic, anti-hyperglycemic, anti-asthmatic and much more. Despite great medicinal worth, little genomic information is available on Aloe vera. This study is an initiative to explore the full-scale functional genomics of Aloe vera by generating whole transcriptome sequence database, using Illumina HiSeq technology and its progressive annotation specifically with respect to the metabolic specificity of the plant.ResultsTranscriptome sequencing of root and leaf tissue of Aloe vera was performed using Illumina paired-end sequencing technology. De novo assembly of high quality paired-end reads, resulted into 1,61,733 and 2,21,792 transcripts with mean length of 709 and 714 nucleotides for root and leaf respectively. The non-redundant transcripts were clustered using CD-HIT-EST, yielding a total of 1,13,063 and 1,41,310 unigenes for root and leaf respectively. A total of 6114 and 6527 CDS for root and leaf tissue were enriched into 24 different biological pathway categories using KEGG pathway database. DGE profile prepared by calculating FPKM values was analyzed for differential expression of specific gene encoding enzymes involved in secondary metabolite biosynthesis. Sixteen putative genes related to saponin, lignin, anthraquinone, and carotenoid biosynthesis were selected for quantitative expression by real-time PCR. DGE as well as qRT PCR expression analysis represented up-regulation of secondary metabolic genes in root as compared to leaf. Furthermore maximum number of genes was found to be up-regulated after the induction of methyl jasmonate, which stipulates the association of secondary metabolite synthesis with the plant’s defense mechanism during stress. Various transcription factors including bHLH, NAC, MYB were identified by searching predicted CDS against PlantTFdb.ConclusionsThis is the first transcriptome database of Aloe vera and can be potentially utilized to characterize the genes involved in the biosynthesis of important secondary metabolites, metabolic regulation, signal transduction mechanism, understanding function of a particular gene in the biology and physiology of plant of this species as well as other species of Aloe genus.

Quantification of Genomic DNA of 125 Chickpea (Cicer Arietinum L.) Genotypes

Chickpea (Cicer arietinum L., 2n=16) is an autogamous annual cool season grain legume cultivated in arid and semi-arid areas across the world.1 It is the third most important food legume in the world after dry beans and peas.2 Chickpea was one of the first grain legumes to be domesticated in the old world.3 This primordial crop probably originated 7000 years ago in an area of present-day south-eastern Turkey and adjoining areas of Syria, and spread from there to Middle East, South Asia and North Africa, where it become an imperative crop. The plant of chickpea stands between 20cm and 1m tall,4 with a genome size of 740Mbp,5,6 slightly less than the well-characterized tomato genome (950Mbp). Nearly 90% of the crop is cultivated under rainfed conditions mostly on receding soil moisture. Chickpea being a rabi crop is normally sown in the month of October and harvested in March, whereas in northeastern Australia, it is sown in May/ June and harvested from October to December.7 Chickpea has high nutritive value and serves as an important cheap source of protein in developing countries diet in addition to improving land fertility.8 The main use of chickpea seeds is for human consumption, especially for strictly vegetarian people, as they are free from any anti-nutritional factors and are rich in phosphorus, calcium and digestible proteins.9 They may be consumed whole as dhal/flour, or the juvenile shoots may be eaten as a vegetable.4 It is the most economical and easily available source of carbohydrates (57-60%), protein (19.5%), fats (1.4%), moisture (4.9-15.59%) and ash (4.8%)10 and it is a good source of calcium, magnesium, potassium, phosphorus, iron, zinc and manganese.11 Utilization of diverse germplasm is required to enhance the genetic variability of cultivars. The major objectives of chickpea breeding are to increase yield either by upgrading the genetic potential or by eliminating the effects of disease, pests and stress.12 Opportunity to generate favourable gene combinations can be provided by genetically diverse lines; thus, the probability of creating unique cultivars increases.13 Therefore, in the present study we extract and quantify the genomic DNA of 125 chickpea (cicer arietinum L.) genotypes. Materials and method