PROTEINASE INHIBITORS

Tyrosinase has an important role in melanin formation, is responsible for the production of colour pigments of skin, hair, and eye. In the presents study, tyrosinase was isolated from Mushrooms, isolation of enzyme was done by acetone precipitation procedure and precipitation of enzyme was done with ammonium sulphate precipitation method. Plants selected for extraction were Azadirachta indica (Neem), Manikara zapota (Chiku), Annona squamosa (Sitaphal), and Hibiscus Rosa-sinesis (China rose). For phytochemical screening Alkaloids-Mayer's Test, Flavonoids (Shinoda Test, Alkaline Reagent Test), sugar (Benedict's reagent Test), Glycosides (Borntrager's Test), Phenolic compounds Test (Ferric chloride Test, Gelatin Test, Lead Acetate Test). Mushroom tyrosinase inhibitory assay was determined by the spectroscopic method. The study shows the tyrosinase inhibitory activity of selected medicinal plants.

Исследование проведено у трех групп больных (45 человек) острым катаральным риносинуситом с применением в комплексе лечения мукоактивных препаратов. Установлено, что данная патология характеризуется острофазным повышением антитрипсиновой активности в сыворотке крови и в синоназальном секрете. При сравнительном анализе результатов применения мукоактивных препаратов различных групп показано, что при данной форме воспаления терапевтический эффект выше, если использовать препарат BNO-101, который на фоне нормализации клинических симптомов приводил к снижению активности эластазо-и трипсиноподобных протеаз, а также к нормализации активности ингибиторов протеиназ в синоназальном секрете. Библиогр. 12 назв. Табл. 1. Ключевые слова: острое катаральное воспаление, риносинусит, мукоактивные препараты, неспецифические протеиназы, ингибиторы неспецифических протеиназ.

The prime purpose of the Market Segment Lead (MSL) is ensuring the development of superior acrosscrops portfolio that will address grower/ customer needs based on Target Product Profile developed in conjunction with the business functions. The MSL function will represent the Seeds Development (SD)organization in the market and be responsible for translating market needs and business plan into resource and strategy. Manage performance, priorities and career development of all members of the MST. Collaborate with SD functions and portfolio managers to develop resource plan that will support the business objectives for the region. Perform annual quantitative pipeline assessment to support the investment in certain crop/ region. • Develop resource plan and budget forecast (excl. budget management) to support market needs. This includes seed shipments, compliance, grower/ field contracts, labour and other activities aiming to secure product development in the market across all crops. • Engage with the budget planning process, pipeline review and 5Y plan exercise.

0The fall armyworm (Spodoptera frugiperda) is an important pest insect due to high degree of polyphagia. In order to better understand its adaptation mechanism against plant protease inhibitors, bioassays were carried supplementing diet with the Kunitz trypsin inhibitor from Entada acaciifolia seeds (EATI). In vitro assays showed an increase of proteolytic activity in EATI-fed larvae midgut. Moreover, the trypsin enzymes showed insensitivity to inhibition with EATI. In order to understand what genes were overexpressed after chronic exposition to EATI, quantitative RT-PCR analyses were performed and revealed an increase in transcription of two trypsin genes, suggesting its participation in insensitivity of midgut trypsins. Another important result was the expression of one chymotrypsin gene, which is not expressed in control fed-larvae but induced in EATI-fed larvae. New regions of higher molecular weight showing proteolytic activity were visualized in inhibitor-fed larvae by zymography gel electrophoresis, proposing that the new enzymes expressed in response of inhibitor dietary would be formatting oligomers. This is a characteristic also observed in other pest insects that adapt to feed in plant protease inhibitors diet. Additional assays revealed that trypsins from EATI-fed larvae also became insensitive against Kunitz and Bowman-Birk inhibitors from soybean. This result suggests a possible involvement of the same S. frugiperda genes in adaptation against Kunitz and Bowman-Birk inhibitors in their host plants.

A new trypsin inhibitor (EATI) was isolated from Entada acaciifolia (Benth.) seeds. EATI is a competitive inhibitor with a molecular mass of 20 kDa and an inhibition stoichiometry of 1:1 for bovine trypsin. The dissociation constant (K i) calculated was 1.75 nmol/L, displaying a high affinity between enzyme and inhibitor. Both Native PAGE and RP-HPLC revealed that EATI is composed of four isoinhibitors that share the amino acid composition and the amino-terminal sequence homolog to Kunitz-type inhibitors. EATI is stable to denaturation by heat (up to 70 • C), pH (2-10), urea (8 mol/L) and its inhibitory activity was unaltered in different concentrations of DTT (up to 100 mmol/L). CD analysis revealed that EATI in reduced form underwent structural modifications associated with a decrease in thermal and pH stabilities, suggesting that their disulfide bonds are not involved in the structuring of its reactive site, but are important for maintenance of its conformational stability. This behavior makes EATI one of the few inhibitors described in the literature with high DTT resistance.

A new trypsin inhibitor (EATI) was isolated from Entada acaciifolia (Benth.) seeds. EATI is a competitive inhibitor with a molecular mass of 20 kDa and an inhibition stoichiometry of 1:1 for bovine trypsin. The dissociation constant (K i) calculated was 1.75 nmol/L, displaying a high affinity between enzyme and inhibitor. Both Native PAGE and RP-HPLC revealed that EATI is composed of four isoinhibitors that share the amino acid composition and the amino-terminal sequence homolog to Kunitz-type inhibitors. EATI is stable to denaturation by heat (up to 70 • C), pH (2-10), urea (8 mol/L) and its inhibitory activity was unaltered in different concentrations of DTT (up to 100 mmol/L). CD analysis revealed that EATI in reduced form underwent structural modifications associated with a decrease in thermal and pH stabilities, suggesting that their disulfide bonds are not involved in the structuring of its reactive site, but are important for maintenance of its conformational stability. This behavior makes EATI one of the few inhibitors described in the literature with high DTT resistance.

Potato leaves infected with Phytophthora infestans produced a serine protease inhibitor (PLPKI) with specificity for microbial proteases. Sequencing of the first twenty residues at the NH 2-terminus of the mature PLPKI polypeptide demonstrated that PLPKI is a novel member of the potato protease inhibitor I family. PLPKI inhibited the activity of extracellular proteases produced by two pathogens of potato, P. infestans and Rhizoctonia solani, but was inactive against proteases secreted into the culture media by the binucleate Rhizoctonia N2, a non-pathogenic fungus for potato. Western blot analysis showed a positive correlation between the levels of PLPKI and the degree of horizontal resistance, showing its highest accumulation in clone OKA 5632.11, which has been described as highly resistant. This correlation, together with the ability of PLPKI to completely abolish the secreted serine protease activity of P. infestans, suggests that PLPKI may have an active role in protecting potato plants from this pathogenic oomycete and that it could be used as a suitable biochemical marker to help breeders in the selection of cultivars with high degree of horizontal resistance.

American Trypanosomiasis or Chagas disease is a prevalent, neglected and serious debilitating illness caused by the kinetoplastid protozoan parasite Trypanosoma cruzi. The current chemotherapy is limited only to nifurtimox and benznidazole, two drugs that have poor efficacy in the chronic phase and are rather toxic. In this scenario, more efficacious and safer drugs, preferentially acting through a different mechanism of action and directed against novel targets, are particularly welcome. Cruzipain, the main papain-like cysteine peptidase of T. cruzi, is an important virulence factor and a chemotherapeutic target with excellent pre-clinical validation evidence. Here, we present the identification of new Cruzipain inhibitory scaffolds within the GlaxoSmithKline HAT (Human African Trypanosomiasis) and Chagas chemical boxes, two collections grouping 404 non-cytotoxic compounds with high antiparasitic potency, drug-likeness, structural diversity and scientific novelty. We have adapted a...

Potato leaves infected with Phytophthora infestans produced a serine protease inhibitor (PLPKI) with specificity for microbial proteases. Sequencing of the first twenty residues at the NH 2-terminus of the mature PLPKI polypeptide demonstrated that PLPKI is a novel member of the potato protease inhibitor I family. PLPKI inhibited the activity of extracellular proteases produced by two pathogens of potato, P. infestans and Rhizoctonia solani, but was inactive against proteases secreted into the culture media by the binucleate Rhizoctonia N2, a non-pathogenic fungus for potato. Western blot analysis showed a positive correlation between the levels of PLPKI and the degree of horizontal resistance, showing its highest accumulation in clone OKA 5632.11, which has been described as highly resistant. This correlation, together with the ability of PLPKI to completely abolish the secreted serine protease activity of P. infestans, suggests that PLPKI may have an active role in protecting potato plants from this pathogenic oomycete and that it could be used as a suitable biochemical marker to help breeders in the selection of cultivars with high degree of horizontal resistance.

Defensive proteins, such as polyphenol oxidase (PPO) and trypsin inhibitor (TI), are induced by herbivore wounding and exogenous methyl jasmonate application in various plant species. This study was conducted to measure induction of PPO and TI in radish, sweet pepper, tomato, and water spinach plants following herbivore wounding (I), methyl jasmonate application (M), and a combination of the two treatments (M + I). The effect of induced responses was also examined against third instar Spodoptera litura Fab. PPO activity was induced in radish by treatment I only; in sweet pepper, by treatments I and M; in tomato, by treatments I, M, and M + I; and in water spinach, by treatments M and M + I. The activity of TI was enhanced 1.2-1.4-fold in radish, sweet pepper, and tomato by M and M + I treatments, whereas in water spinach, it was enhanced 1.2-fold by all 3 treatments. The relative growth rate (RGR) of S. litura was reduced by 53% on radish plants following M treatment only. It was reduced by 37% and 42% on sweet paper plants following M and M + I treatment, respectively. RGR was significantly reduced on test tomato plants following I, M, and M + I treatments. The RGR of S. litura was unaffected on water spinach plants following any treatment. Collectively, the results of this study indicated that induction of plant defensive proteins in response to S. litura feeding or exogenous methyl jasmonate application varied among plant species, which further affected the induced plant resistance to the caterpillars.

Potato leaves infected with Phytophthora infestans produced a serine protease inhibitor (PLPKI) with specificity for microbial proteases. Sequencing of the first twenty residues at the NH 2-terminus of the mature PLPKI polypeptide demonstrated that PLPKI is a novel member of the potato protease inhibitor I family. PLPKI inhibited the activity of extracellular proteases produced by two pathogens of potato, P. infestans and Rhizoctonia solani, but was inactive against proteases secreted into the culture media by the binucleate Rhizoctonia N2, a non-pathogenic fungus for potato. Western blot analysis showed a positive correlation between the levels of PLPKI and the degree of horizontal resistance, showing its highest accumulation in clone OKA 5632.11, which has been described as highly resistant. This correlation, together with the ability of PLPKI to completely abolish the secreted serine protease activity of P. infestans, suggests that PLPKI may have an active role in protecting potato plants from this pathogenic oomycete and that it could be used as a suitable biochemical marker to help breeders in the selection of cultivars with high degree of horizontal resistance.

Defensive proteins, such as polyphenol oxidase (PPO) and trypsin inhibitor (TI), are induced by herbivore wounding and exogenous methyl jasmonate application in various plant species. This study was conducted to measure induction of PPO and TI in radish, sweet pepper, tomato, and water spinach plants following herbivore wounding (I), methyl jasmonate application (M), and a combination of the two treatments (M + I). The effect of induced responses was also examined against third instar Spodoptera litura Fab. PPO activity was induced in radish by treatment I only; in sweet pepper, by treatments I and M; in tomato, by treatments I, M, and M + I; and in water spinach, by treatments M and M + I. The activity of TI was enhanced 1.2-1.4-fold in radish, sweet pepper, and tomato by M and M + I treatments, whereas in water spinach, it was enhanced 1.2-fold by all 3 treatments. The relative growth rate (RGR) of S. litura was reduced by 53% on radish plants following M treatment only. It was reduced by 37% and 42% on sweet paper plants following M and M + I treatment, respectively. RGR was significantly reduced on test tomato plants following I, M, and M + I treatments. The RGR of S. litura was unaffected on water spinach plants following any treatment. Collectively, the results of this study indicated that induction of plant defensive proteins in response to S. litura feeding or exogenous methyl jasmonate application varied among plant species, which further affected the induced plant resistance to the caterpillars.

Protease inhibitors (PIs) are deployed in the plant kingdom as storage proteins or peptides, regulators of endogenous proteases, and plant protection agents against insect pests and pathogen attack. In humans, they are identified as chemopreventive agents against a range of cancers and have potential as drug to treat an array of disease associated with aberrant activity of proteases. The present investigation reports PIs activity data from 30 medicinal plants. The screening for PIs activity was done by dot blot assay using X-ray film coated with gelatin. Among screened seed extracts, Albizia lebbeck, Raphanus sativus, Mucuna pruriens, Achyranthes aspera, and Coffea arabica showed high inhibitory activities with trypsin protease. Most of seed extracts exhibited moderate activity, whereas Ocimum sanctum showed moderate to low activity against trypsin. The presence of varied protein content is reported from all seed extracts with highest in A. lebbeck (50.0 ± 3.4 mg/ml). The data produ...

An artificial diet for mass rearing of the tobacco caterpillar Spodoptera litura (Fabricius) from the neonate to adult stage was developed at 27^18C, 65^5% RH and 16:8 h scoto/photo-phase regime. The diet ingredients consisted of wheat germ (26.0 g), kidney bean flour (51.3 g), chickpea flour (56.0 g), yeast powder (31.6 g), casein (15.2 g), L-ascorbic acid (3.2 g), cholesterol (0.5 g), two multivitamin multimineral capsules, one vitamin E capsule, ABDEC drops (2 ml), castor oil (1 ml), methyl-p-hydroxybenzoate (1.8 g), sorbic acid (1.3 g), streptomycin sulphate (0.25 g), formaldehyde solution (2 ml), agar-agar (16.4 g) and 820 ml distilled water. The new artificial diet successfully supported the growth and development for more than ten generations with enhanced reproductive potential. The mean biological parameters based on rearing of ten continuous generations showed higher pupation (89.2%), emergence (97.2%), survival (86.6%) and fecundity (2486.2 eggs) as compared to the most preferred natural food, castor leaf whose respective recorded values are 80, 75, 60% and 480 eggs. The cost of 1 l of the diet was approximately US$ 2.00 only, on which 200 neonate larvae can be reared. The artificial diet developed is suitable for mass rearing S. litura successfully throughout the year for laboratory and field experimentation as well as for commercial production of nuclear polyhedrosis viruses (Sl NPV).

Damage to foliage of the tomato, Lycopersicon esculentum, causes the induction of proteinase inhibitors and of the oxidative enzymes polyphenol oxidase, peroxidase, and lipoxygenase. The time courses of induction of these proteins by feeding of two caterpillar species (Manduca sexta and Helicoverpa zea) were studied in a series of experiments. In another series of experiments, the effects of plant age on the inducibility of these proteins were studied. In the time course experiments, induction of proteinase inhibitors and oxidative enzymes in the damaged leaflet was rapid, with higher protein activities evident in damaged leaflets within 12-24 h of damage, depending on the enzyme and the species of insect used to damage the plant. Systemic induction of proteinase inhibitors was also rapid, but systemic induction of polyphenol oxidase was delayed relative to systemic induction of proteinase inhibitors, possibly because high constitutive polyphenol oxidase activities obscured expression of systemic induction at earlier time points. Lipoxygenase and peroxidase were not induced systemically. Induction of all proteins persisted for at least 21 days. In the phenology experiments, inducibility of all proteins decreased in magnitude and was less consistent as plants aged. The results of these experiments exemplify the numerous constraints on induction in tomato plants. Knowledge of these physiological constraints is important to an understanding of the ecological role and causal basis of induced resistance.

Sugarcane woolly aphid, Ceratovacuna lanigera is one of the important pests of the sugarcane which is responsible for the low production of the sugarcane in India. The incidences of pest itself suggest importance of study on this pest. Carbohydrate metabolizing digestive enzyme, α-amylase was studied from this insect. Biochemical characterization of this enzyme and screening of natural inhibitors will prove vital strategies against infestation. As initial step to this goal here we report the detection of α-amylase of Ceratovacuna lanigera by starch agar well diffusion assay. Biochemical parameters like pH, temperature and effect of substrate concentration were studied and compared with α-amylases from other sources. The optimum pH of α-amylase of Ceratovacuna lanigera was found to be pH 8 in contrast to human salivary α-amylases and fungal α-amylases those shows optimum activity at pH 7 and pH 6 respectively. Maximum activity of α-amylase was found to be at 40°C. Protein profile by Native-PAGE showed banding pattern with varied intensities. The α-amylase of Ceratovacuna lanigera detected in the present study needs further exploration for its use in better management of sugarcane woolly aphid.

Here, we report multiple molecular forms of Albizia lebbeck trypsin inhibitors (AlTIs) by using a simple and sensitive gel X-ray film contact print technique. About 17 AlTIs were detected in the seed extracts of A. lebbeck. Two groups of AlTIs-1 major (10 AlTIs; slow migration on the gel) and 1 minor (7 AlTIs; fast migration on the gel) were identified. The former was specific only toward trypsin. However, the latter was specific toward both trypsin and Helicoverpa armigera gut proteinases (HaGPs). The most potent AlTI (AlTI 13) was purified to assess its in vivo bioefficacy toward HaGPs. Purification was achieved using (NH 4) 2 SO 4 fractionation, Sephadex G-100 column chromatography, and preparative native-polyacrylamide gel electrophoresis (PAGE). The dose dependent bioefficacies of AlTIs in the (NH 4) 2 SO 4 F 3 fractions (0.1%, 0.5%, and 1%) were approximately 79%, 83%, and 90%, respectively, resulting in reductions in the average larval weight of H. armigera. Artificial diet containing a single dose of AlTI 13 (5 μg/g diet) reduced the larval weight by about 76%, with 60% mortality. The half-maximal inhibitory concentrations (IC 50) of AlTI 13 for trypsin and HaGPs were 0.14 and 0.17 μmol/ml, respectively. The optimum conditions for AlTI 13 were pH 8 and temperatures ranging from 35 to 40°C. Reducing sodium dodecyl sulfate-PAGE analysis indicated that~28 kDa Kunitz-like trypsin inhibitor was present. Thus, we showed that AlTIs, particularly, AlTI 13 of A. lebbeck could be used as a transgene macromolecule to markedly increase insect resistance in genetically engineered plants.

Heliothis amigera was reared on four varieties of gram, namely Radhe, Desi, K,,, and K,. Growth was studied under laboratory conditions from January to March 1986. The weight acquired by the mature larvae showed variation among the tested food plants. Weight of the pupa also varied in response to rearing on different varieties of gram. Radhe induced the quickest pupation. The emergence of the insect varied from 61-92% among the tested varieties of gram. Fecundity also differed markedly from variety to variety.

The gene of the 85 kDa cysteine protease inhibitor (CPI) potato multicystatin (PMC), was PCR-cloned and expressed under the control of the UEP1 promoter in chrysanthemum (Dendranthema grandiflora). Over 30 independent transgenic plant lines were analysed for PMC expression in florets by immunoassay and a selection of those for papain inhibitor activity (PIA). A significant correlation between PIA activity and PMC expression levels was established demonstrating that the highest expressers raised the concentration of PIA to 0.28 pmol/µg from an endogenous background level of 0.15 pmol/µg. On this basis, the PMC gene was estimated to be expressed at a level of 0.13% of total protein. Some of the transgenic lines exhibited up to 5-fold higher levels of PIA (0.82 pmol/µg protein). This did not correlate with the immunological data, however, and may be the result of frequently occurring somaclonal variation. A non-choice bioassay of 7-10 days on whole flowers was carried out to study the effect of PMC on the fecundity of Western Flower Thrips in terms of the number of larvae produced. No correlation between the reduction in the oviposition rate and PMC expression could be established, which may be due to the relatively low expression level of PMC in chrysanthemum. The transgenic lines with the highest levels of endogenous PIA also had the lowest thrips reproduction rates, but further experiments are required to exclude artefacts caused by t he aberrant phenotype.

The gene of the 85 kDa cysteine protease inhibitor (CPI) potato multicystatin (PMC), was PCR-cloned and expressed under the control of the UEP1 promoter in chrysanthemum (Dendranthema grandiflora). Over 30 independent transgenic plant lines were analysed for PMC expression in florets by immunoassay and a selection of those for papain inhibitor activity (PIA). A significant correlation between PIA activity and PMC expression levels was established demonstrating that the highest expressers raised the concentration of PIA to 0.28 pmol/µg from an endogenous background level of 0.15 pmol/µg. On this basis, the PMC gene was estimated to be expressed at a level of 0.13% of total protein. Some of the transgenic lines exhibited up to 5-fold higher levels of PIA (0.82 pmol/µg protein). This did not correlate with the immunological data, however, and may be the result of frequently occurring somaclonal variation. A non-choice bioassay of 7-10 days on whole flowers was carried out to study the effect of PMC on the fecundity of Western Flower Thrips in terms of the number of larvae produced. No correlation between the reduction in the oviposition rate and PMC expression could be established, which may be due to the relatively low expression level of PMC in chrysanthemum. The transgenic lines with the highest levels of endogenous PIA also had the lowest thrips reproduction rates, but further experiments are required to exclude artefacts caused by t he aberrant phenotype.

Area of genomics revolves around the study of genes and gene functioning. It is broadly sub categorized as structural genomics, functional genomics, comparative genomics, epigenomics, translational genomics and pharmacogenomics. Functional genomics, highlighted in this account, links the organism’s genome to its function and phenotype. It could be further subdivided into proteomics, transcriptomics and metabolomics and could be studied by reverse genomic echniques like RNAi and mutagenesis. In agriculture, functional genomics finds its application in generation of crop plants with highly improved agronomic traits so that the exponentially increasing population is well fed, today and in near future. With high through-put and next generation approaches, functional genomics might help in circumventing hindrance in achieving food security. Though crucial, this approach involves meticulous planning and efficient policy making along with brotherhood from whole scientific fraternity in ach...

The present investigation on the impact of fungicides on the growth and development of Spodoptera litura was carried out in the laboratory of the Department of Entomology, SKUAST-Jammu, during the year 2012-13. Base-line toxicity of two fungicides viz., mancozeb and ridomil MZ were evaluated against S. litura in the laboratory. The results showed that fungicides significantly influenced the growth and development of S. litura. The larval development duration was significantly short, and it was an average (14.61± 0.30), (16.28±0.66) days, when treated with ridomil, and mancozeb respectively. The results show that fungicides can serve a practical tool to reduce the S.litura and may assume a greater role in integrated programs showed to manage insect pests and pathogens.

Abstract: This paper focuses on India's emerging role in global bioinformatics market. Bioinformatics is growing as an independent discipline and is fundamental to the growth of biotechnology. India has achieved remarkable success in the software industry. It would add nine per cent to India's GDP by 2010. With India's strong presence in world software market, India is expected to play a leading role in the bioinformatics revolution. This paper highlights Indian and global bioinformatics scenario. It also discusses bioinformatics challenges and ...

Plant protease inhibitors (PPIs) are compounds which play a potent defensive role against pest and pathogens, thereby protecting the plants. In this work we tested different plant extracts to identify extracts containing inhibitor against trypsin activity. Plant extracts were made by homogenizing soaked leaves/seeds/root modification of plants in bicarbonate buffer, pH 9.0.The homogenates were centrifuged at 10,000 rpm at 4°C for 10 minutes. The supernatant containing the soluble proteins were used for protease inhibition assay. Protease inhibition assay was carried out in a total volume of 1 ml containing 67 µl trypsin (Bovine), 25 µl plant extract, 511 µl Tris buffer, 67µl NaCl and 330 µg Nα-Benzoyl-DL-Arginine-P-Nitro Anilide (BAPNA) as substrate. On screening 19 extracts from different plants, we found 9 of them inhibited trypsin activity greater than 40%. The percentage inhibition of these plants are Garcinia xanthochymus (96.7± 0.4 %), Datura stramonium (86.1±1.6%), Ricinus co...

The cotton bollworm, Helicoverpa armigera Hubner (Lep: Noctuidae), is an economically important pest of numerous major food crops worldwide. Protease inhibitors from plants, expressed constitutively in transgenic crops, have potential for pest management as an alternative to chemical pesticides. In this study, a protease inhibitor was isolated, purified, and characterized from Datura metel L. seeds. The purity of the isolated inhibitor was confirmed by reverse-phase high-performance liquid chromatography, and activity staining showed one major peak and one clear activity band for the protein. Electrophoretic studies following gel filtration and ion-exchange chromatography revealed two and one bands for purified proteins, respectively. Partial biochemical characterizations of the purified inhibitor were determined. Maximum inhibitory activity was observed at 40–45°C (optimal temperature) when tested against gut extracts of fourth to sixth instar H. armigera larvae.Thermo-stability of...

Protease inhibitors (PIs) are deployed in the plant kingdom as storage proteins or peptides, regulators of endogenous proteases, and plant protection agents against insect pests and pathogen attack. In humans, they are identified as chemopreventive agents against a range of cancers and have potential as drug to treat an array of disease associated with aberrant activity of proteases. The present investigation reports PIs activity data from 30 medicinal plants. The screening for PIs activity was done by dot blot assay using X-ray film coated with gelatin. Among screened seed extracts, Albizia lebbeck, Raphanus sativus, Mucuna pruriens, Achyranthes aspera, and Coffea arabica showed high inhibitory activities with trypsin protease. Most of seed extracts exhibited moderate activity, whereas Ocimum sanctum showed moderate to low activity against trypsin. The presence of varied protein content is reported from all seed extracts with highest in A. lebbeck (50.0 ± 3.4 mg/ml). The data produced in the present investigation could be helpful for further exploration of PIs as therapeutic agent.

Protease inhibitors (PIs) are deployed in the plant kingdom as storage proteins or peptides, regulators of endogenous proteases, and plant protection agents against insect pests and pathogen attack. In humans, they are identified as chemopreventive agents against a range of cancers and have potential as drug to treat an array of disease associated with aberrant activity of proteases. The present investigation reports PIs activity data from 30 medicinal plants. The screening for PIs activity was done by dot blot assay using X-ray film coated with gelatin. Among screened seed extracts, Albizia lebbeck, Raphanus sativus, Mucuna pruriens, Achyranthes aspera, and Coffea arabica showed high inhibitory activities with trypsin protease. Most of seed extracts exhibited moderate activity, whereas Ocimum sanctum showed moderate to low activity against trypsin. The presence of varied protein content is reported from all seed extracts with highest in A. lebbeck (50.0 ± 3.4 mg/ml). The data produced in the present investigation could be helpful for further exploration of PIs as therapeutic agent.

This paper focuses on India’s emerging role in global bioinformatics market. Bioinformatics is growing as an independent discipline and is fundamental to the growth of biotechnology. India has achieved remarkable success in the software industry. It would add nine per cent to India’s GDP by 2010. With India’s strong presence in world software market, India is expected to play a leading role in the bioinformatics revolution. This paper highlights Indian and global bioinformatics scenario. It also discusses bioinformatics challenges and opportunities and examines India’s competitive position in bioinformatics by using S.W.O.T. analysis framework.

Bioinformatics: Meaning and Definitions Bioinformatics is a highly interdisciplinary, using techniques and concepts from applied mathematics, informatics, statistics, chemistry, biochemistry, physics, linguistics and computer science to solve biological problems and is the interface between the biological and computational sciences. Bioinformatics is an integration of mathematical, statistical and computer methods to analyze biological, biochemical and biophysical data. It is the application of computer technology to the management and analysis of biological data such as the sequences of DNA (Deoxyribonucleic acid) and RNA (Ribonucleic acid) in genes and amino acids in proteins.

The gene of the 85 kDa cysteine protease inhibitor (CPI) potato multicystatin (PMC), was PCR-cloned and expressed under the control of the UEP1 promoter in chrysanthemum (Dendranthema grandiflora). Over 30 independent transgenic plant lines were analysed for PMC expression in florets by immunoassay and a selection of those for papain inhibitor activity (PIA). A significant correlation between PIA activity and PMC expression levels was established demonstrating that the highest expressers raised the concentration of PIA to 0.28 pmol/µg from an endogenous background level of 0.15 pmol/µg. On this basis, the PMC gene was estimated to be expressed at a level of 0.13% of total protein. Some of the transgenic lines exhibited up to 5-fold higher levels of PIA (0.82 pmol/µg protein). This did not correlate with the immunological data, however, and may be the result of frequently occurring somaclonal variation. A non-choice bioassay of 7-10 days on whole flowers was carried out to study the effect of PMC on the fecundity of Western Flower Thrips in terms of the number of larvae produced. No correlation between the reduction in the oviposition rate and PMC expression could be established, which may be due to the relatively low expression level of PMC in chrysanthemum. The transgenic lines with the highest levels of endogenous PIA also had the lowest thrips reproduction rates, but further experiments are required to exclude artefacts caused by t he aberrant phenotype.

Protease inhibitors (PIs) are deployed in the plant kingdom as storage proteins or peptides, regulators of endogenous proteases, and plant protection agents against insect pests and pathogen attack. In humans, they are identified as chemopreventive agents against a range of cancers and have potential as drug to treat an array of disease associated with aberrant activity of proteases. The present investigation reports PIs activity data from 30 medicinal plants. The screening for PIs activity was done by dot blot assay using X-ray film coated with gelatin. Among screened seed extracts, Albizia lebbeck, Raphanus sativus, Mucuna pruriens, Achyranthes aspera, and Coffea arabica showed high inhibitory activities with trypsin protease. Most of seed extracts exhibited moderate activity, whereas Ocimum sanctum showed moderate to low activity against trypsin. The presence of varied protein content is reported from all seed extracts with highest in A. lebbeck (50.0 ± 3.4 mg/ml). The data produ...

Proteolytic activity in whole insect extracts of the western flower thrips, Frankliniella occidentalis, was found to belong predominantly to the class of cysteine proteases. The pH optimum of the general proteolytic activity was determined to be 3.5, which is low when compared to other insects using cysteine proteases for protein digestion. The proteinaceous cysteine protease inhibitors chicken cystatin, potato cystatin and sea anemone equistatin inhibited in vitro more than 90% of the protease activity. To test in vivo the biological effect of such inhibitors on the oviposition rate of western flower thrips, recombinant potato cystatin and equistatin were fed to adult females. A gradual reduction in oviposition rate to about 45% of control was observed when reared on these PIs for a period of 5 days, with no increase in mortality. These results are discussed in the light of the application of protease inhibitors in transgenic plants to control this insect pest.

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Serine proteinase inhibitors, produced by plants, inhibiting protein digestion in herbivorous insects, can be used as one of the best ecofriendly IPM tactics. In this context, laboratory assay was conducted to identify the potential source of serine proteinase inhibitors from the seeds of 38 plant species corresponding to nine families. Among them, extracts from 20 species exhibited both trypsin and chymotrypsin inhibitory activity (TIA and CIA respectively) and 15 plant species possess TIA. The maximum TIA [9878 TIU (Trypsin Inhibitory Unit) g-1 of seed] was observed in Momordica balsamina seed extract. Erythrina variegata was found to possess both TIA (2737 TIU g-1 of seed) and CIA (2800 CIU Chymotrypsin Inhibitory Unit g-1 of seed). Seeds of two species may be utilized as a potential source of proteinase inhibitor in insect control programme. These presence of trypsin inhibitory activity in M. balsamina is reported for the first time.

Because the current HIV-1 therapy takes a high cost and associates with the appearance of multi-drug resistant HIV-1 strain, Indian traditional Ayurveda might become useful alternative. In this study, we screened for inhibitory activities of various extracts of five Indian plants for HIV-1 reverse transcriptase (RT). Water-extracts of leaf of Argemone mexicana strongly inhibited the DNA polymerase activity of HIV-1 RT, indicating that they contain substances that inhibit the activity. Neither thermal treatment at 100ºC nor proteinase K treatment of the extracts abolished the inhibitory activity, suggesting that the inhibitory substance is an organic compound rather than a protein.

Antimetabolic effect of purified proteinase inhibitor (F30-60) isolated from Adenanthera pavonina (ApPI F30-60) was evaluated on Spodoptera litura larva, by feeding five days old larva on the ApPI F30-60contaminated diet at 0.25, 0.50 and 1.00% for acute period (5 days) and later fed with untreated diet. The parameters of growth and development of S. litura were closely monitored throughout its life period. The results indicated that the ApPI 1.0% had significant influence on the reduction of S. litura larval weight(41.58%) pupal and adult weight (38.42 and 44.42%, respectively), prolonged the larval and pupal developmental period by 7.2 days and 3.0 days, respectively reduced the adult life span by 4.5 days. Only 60 per cent of the treated larva were pupated, 30 per cent of them were malformed as pupal-adult intermediates. Hence only 30 per cent were emerged as adult but they did not lay eggs. Acute feeding of ApPI 1% significantly interfered in the growth and development of S. litura. Hence, it is concluded that purified A. pavonina proteinase inhibitor (ApPI) is a potential and promising inhibitor of both chymotrypsin and trypsin in S. litura, which can be effectively exploited as bio insecticidal tool against this notorious polyphagous pest on a range of field and horticultural crops.

The potential trypsin inhibitor sources were identified from seeds of ten plants, based on the reaction of trypsin with synthetic substrate Nα-Benzoyl-D, L-arginine 4-nitroanilide hydrochloride (BApNA). All the tested ones contained various levels of protein (20.87 to 155.68 mg g-1 of seed) and trypsin inhibitor units (TIU) (245.77 to 11235.82 TIU g-1 seed). Among them, the Red lucky seed (Adenanthera pavonina L.) was identified as the richest source of trypsin inhibitor with 11235.82 TIU and 96.06 mg protein/ g of seed with specific activity of 116.97 TIU mg-1 protein. The next best ones the seeds of bitter gourd (Momordica charantia L.) and babul (Acacia nilotica L.) with the TIU g-1 seed as 9100.03 and 8690.72, protein content of 155.68 and 88.11 mg g-1 seed with the specific activity of 58.45 and 98.63 TIU mg-1 protein, respectively. This study is the first report of Zingiber officinale Rosc. as the source of trypsin inhibitor.

In the present study, twelve varieties of CHRYSANTHEMUM collected from the National Botanical Research Institute, Lucknow were evaluated under irrigated condition at Main Experiment Station, NDUA&T, Kumarganj, Faizabad during winter season (2011-12). A wide range of variation in the performance of the varieties were observed for various characters. Highest plant height (50.20 cm) and maximum number of florets per flower were observed in Suneel (339.67 cm). Genotype Dentiment showed maximum plant spread (34.74 cm), maximum number of flowers per plant (103.60 g), maximum flower diameter (10.40 cm), and highest weight of ten flowers (63.00 g). As for as suitability of particular genotypes is concerned, maximum number of primary branches in Jaya (10.30) and maximum flower vase life (16 days) in Jayanti was observed. Among all the 12 genotypes, few genotypes like Dentiment produced maximum flower yield (652.68 g per plant) Original Research Article

The inhibitory effects of human a 2 -macroglobulin (a 2 -M), a major plasma proteinase inhibitor, on human pepsin and gastricsin were investigated. The activities of pepsin and gastricsin towards a protein substrate (reduced and carboxymethylated ribonuclease A) were significantly inhibited by a 2 -M at pH 5.5, whereas those towards a peptide substrate (oxidized insulin B-chain) were scarcely inhibited. Under these conditions at pH 5.5, pepsin and gastricsin cleaved a 2 -M mainly at the His 694 -Ala 695 bond and Leu 697 -Val 698 bond, respectively, in the bait regions sequence of a 2 -M. The conformation of a 2 -M was also shown to be markedly altered upon inhibition of these enzymes as examined by native polyacrylamide gel electrophoresis and electron microscopy. These results show the entrapment and concomitant inhibition of those proteinases by a 2 -M.

The inhibitory effects of human alpha2-macroglobulin (alpha2-M), a major plasma proteinase inhibitor, on human pepsin and gastricsin were investigated. The activities of pepsin and gastricsin towards a protein substrate (reduced and carboxymethylated ribonuclease A) were significantly inhibited by alpha2-M at pH 5.5, whereas those towards a peptide substrate (oxidized insulin B-chain) were scarcely inhibited. Under these conditions at pH 5.5, pepsin and gastricsin cleaved alpha2-M mainly at the His694-Ala695 bond and Leu697-Val698 bond, respectively, in the bait regions sequence of alpha2-M. The conformation of alpha2-M was also shown to be markedly altered upon inhibition of these enzymes as examined by native polyacrylamide gel electrophoresis and electron microscopy. These results show the entrapment and concomitant inhibition of those proteinases by alpha2-M.

The inhibitory effects of human a 2 -macroglobulin (a 2 -M), a major plasma proteinase inhibitor, on human pepsin and gastricsin were investigated. The activities of pepsin and gastricsin towards a protein substrate (reduced and carboxymethylated ribonuclease A) were significantly inhibited by a 2 -M at pH 5.5, whereas those towards a peptide substrate (oxidized insulin B-chain) were scarcely inhibited. Under these conditions at pH 5.5, pepsin and gastricsin cleaved a 2 -M mainly at the His 694 -Ala 695 bond and Leu 697 -Val 698 bond, respectively, in the bait regions sequence of a 2 -M. The conformation of a 2 -M was also shown to be markedly altered upon inhibition of these enzymes as examined by native polyacrylamide gel electrophoresis and electron microscopy. These results show the entrapment and concomitant inhibition of those proteinases by a 2 -M.

Proteolytic activity in whole insect extracts of the western flower thrips, Frankliniella occidentalis, was found to belong predominantly
to the class of cysteine proteases. The pH optimum of the general proteolytic activity was determined to be 3.5, which is
low when compared to other insects using cysteine proteases for protein digestion. The proteinaceous cysteine protease inhibitors
chicken cystatin, potato cystatin and sea anemone equistatin inhibited in vitro more than 90% of the protease activity. To test in
vivo the biological effect of such inhibitors on the oviposition rate of western flower thrips, recombinant potato cystatin and
equistatin were fed to adult females. A gradual reduction in oviposition rate to about 45% of control was observed when reared
on these PIs for a period of 5 days, with no increase in mortality. These results are discussed in the light of the application of
protease inhibitors in transgenic plants to control this insect pest.

Abstract
The gene of the 85 kDa cysteine protease inhibitor (CPI) potato multicystatin (PMC), was
PCR-cloned and expressed under the control of the UEP1 promoter in chrysanthemum
(Dendranthema grandiflora). Over 30 independent transgenic plant lines were analysed
for PMC expression in florets by immunoassay and a selection of those for papain
inhibitor activity (PIA). A significant correlation between PIA activity and PMC
expression levels was established demonstrating that the highest expressers raised the
concentration of PIA to 0.28 pmol/mg from an endogenous background level of 0.15
pmol/mg. On this basis, the PMC gene was estimated to be expressed at a level of 0.13%
of total protein. Some of the transgenic lines exhibited up to 5-fold higher levels of PIA
(0.82 pmol/mg protein). This did not correlate with the immunological data, however, and
may be the result of frequently occurring somaclonal variation. A non-choice bioassay of
7-10 days on whole flowers was carried out to study the effect of PMC on the fecundity
of Western Flower Thrips in terms of the number of larvae produced. No correlation
between the reduction in the oviposition rate and PMC expression could be established,
which may be due to the relatively low expression level of PMC in chrysanthemum. The
transgenic lines with the highest levels of endogenous PIA also had the lowest thrips
reproduction rates, but further experiments are required to exclude artefacts caused by the
aberrant phenotype.

Most of the plant protection strategies are focused on selection and application of the natural proteinase inhibitors (PIs) against insect pests. In addition, PIs also play a vital role in medicine for treatment of immunity related diseases. PI activity exists mainly in seeds, leaves and flowers of plants. In search of novel PIs, 135 different plant tissue extracts (leaf, flower and seed) were screened for their PI (trypsin, chymotrypsin and Helicoverpa gut proteinase inhibitors) activities by using dot-blot assays. Most of the plant tissues screened revealed moderate PI activity, few showed low PI activity and very few of them showed strong PI activity against trypsin, chymotrypsin and Helicoverpa gut proteinases. The inhibitory potency of positive samples was further determined by solution assays. Five plants namely Arachis hypogaea, Vigna sinensis, Dolichos lablab, Phaseolus aureus and Cassia siamea showed higher activity which ranged from 22.91 to 58.33 %. Higher activities reco...

The inactivation of Trypanosoma cruzi proteinases by human h2-macroglobulin (h2-M), a major plasma proteinase inhibitor was studied. Evidences regarding the interaction between h2-M and proteolytic enzymes contained in crude cell-free extracts of T. cruzi were derived from electrophoretic and enzymatic assays. The former showed conformational and structural changes ocurring in h2-M, as judged by the appearance of transformed 'fast' form on native PAGE; generation of bands of : 90 kDa on reduced SDS-PAGE and formation of covalent complexes enzyme-inhibitor on SDS-PAGE. On the other hand, the total proteolytic activity on azocasein dropped significantly in the presence of h2-M, although partial activity was still maintained. The proteinases detected as a double band of 44 and 53 kDa on gelatin SDS-PAGE were also inhibited by h2-M. Results suggest that the study of specific interactions between h2-M and T. cruzi-proteinases, probably with cruzipain, could be biologically important in the fate of T. cruzi-infection and Chagas' disease. © 1997 Elsevier Science B.V.

Most of the plant protection strategies are focused on selection and application of the natural proteinase inhibitors (PIs) against insect pests. In addition, PIs also play a vital role in medicine for treatment of immunity related diseases. PI activity exists mainly in seeds, leaves and flowers of plants. In search of novel PIs, 135 different plant tissue extracts (leaf, flower and seed) were screened for their PI (trypsin, chymotrypsin and Helicoverpa gut proteinase inhibitors) activities by using dot-blot assays. Most of the plant tissues screened revealed moderate PI activity, few showed low PI activity and very few of them showed strong PI activity against trypsin, chymotrypsin and Helicoverpa gut proteinases. The inhibitory potency of positive samples was further determined by solution assays. Five plants namely Arachis hypogaea, Vigna sinensis, Dolichos lablab, Phaseolus aureus and Cassia siamea showed higher activity which ranged from 22.91 to 58.33 %. Higher activities recorded in the seed as compare to leaf and flower tissues. Dolichos lablab showed highest PI activity (58.33 %) followed by Cassia siamea (52.08 %). PI activity was found to be distributed unequally in ammonium sulfate (NH 2 SO 4 ) fractions. 8 methods of controlling this pest. Widespread applications of PIs in agriculture and medicine encourage us (leaves, seeds and flowers) to find novel source. Keeping this goal in mind we screened 140 different plant tissue extracts for PI activity and varied inhibition potential of these tissue extracts against trypsin, chymotrypsin and H. armigera gut proteinases (HGP) were reported.

Poor or improper storage facilities of seeds before sowing are responsible for destruction of a large part of our seed banks. Pre-harvest and post-harvest fungal infections destroy the viability and vigour of even good quality seeds during storage. If only 40% of seeds germinate on sowing, it will be a great economic loss indeed. A simple process like pre-sowing seed treatment with aqueous extracts of common wild plants readily available not only enhances better and vigorous growth ,but also can save up to 60% of seeds from damage due to fungal infections.