02410nas a2200337   4500008004100000022001400041245011600055210006900171260009200240300001200332490000700344520123200351653002701583653002301610653001401633653001601647653002501663653001101688653002701699653001301726100002801739700002201767700002101789700002001810700002401830700002301854700002201877700002001899700002601919856012701945       2008                        eng d  a0958-315700aComparison of Metarhizium isolates for biocontrol of helicoverpa armigera (Lepidoptera : Noctuidae) in chickpea0 aComparison of Metarhizium isolates for biocontrol of helicoverpa  a4 PARK SQUARE, MILTON PARK, ABINGDON OX14 4RN, OXON, ENGLANDbTAYLOR & FRANCIS LTDcFEB  a809-8280 v183 a<p>Metarhizium isolates from soil (53) and insect hosts (10) were evaluated for extracellular production of cuticle degrading enzyme (CDE) activities such as chitinase, chitin deacetylase (CDA), chitosanase, protease and lipase. Regression analysis demonstrated the relation of CDE activities with Helicoverpa armigera mortality. On basis of this relation, ten isolates were selected for further evaluation. Subsequently, based on LT(50) of the 10 isolates towards H. armigera, five isolates were selected. Out of these five isolates, three were selected on the basis of higher conidia production (60-75 g/kg rice), faster sedimentation time (ST(50)) (2.3-2.65 h in 0.1% (w/v) Tween 80) and lower LC(50) (1.4- 5.7 x 10(3) conidia/mL) against H. armigera. Finally, three Metarhizium isolates were selected for the molecular fingerprinting using ITS sequencing and RAPD patterning. All three isolates, M34412, M34311 and M81123, showed comparable RAPD patterns with a 935G primer. These were further evaluated for their field performance against H. armigera in a chickpea crop. The percent efficacies with the three Metarhizium isolates were from 65 to 72%, which was comparable to the chemical insecticide, endosulfan (74%).</p>10aappressorium formation10achitin deacetylase10achitinase10achitosanase10aHelicoverpa armigera10aLipase10aMetarhizium anisopliae10aProtease1 aKulkarni, Shuklangi, A.1 aGhormade, Vandana1 aKulkarni, Girish1 aKapoor, Manisha1 aChavan, Santosh, B.1 aRajendran, Armugam1 aPatil, Sanjay, K.1 aShouche, Yogesh1 aDeshpande, Mukund, V.  uhttps://library.ncl.res.in/content/comparison-metarhizium-isolates-biocontrol-helicoverpa-armigera-lepidoptera-noctuidae-002914nas a2200337   4500008004100000022001400041245014400055210006900199260009200268300001200360490000700372520169100379653002702070653002302097653001402120653002502134653002702159653001102186653002702197653001302224100002302237700002802260700002202288700002402310700002102334700002402355700002102379700002002400700002602420856013002446       2008                        eng d  a0958-315700aEffect of repeated in vitro sub-culturing on the virulence of metarhizium anisopliae against helicoverpa armigera (Lepidoptera : Noctuidae)0 aEffect of repeated in vitro subculturing on the virulence of met  a4 PARK SQUARE, MILTON PARK, ABINGDON OX14 4RN, OXON, ENGLANDbTAYLOR & FRANCIS LTDcMAY  a337-3550 v183 a<p>The effect of repeated conidial sub-culturing of Metarhizium anisopliae on its virulence against Helicoverpa armigera (Hbner) was studied. The LT(50) observed against third instar larvae of H. armigera for the first sub-culture was 3.4 days; it increased to 4.5 and 5.6 days for the 20th and the 40th sub-cultures, respectively. The LT50 values after passage of the 40th sub-culture on H. armigera decreased to 4.4 and 3.7 days for the 40th (first in vivo) and the 40th (fifth in vivo) passages, respectively. Similarly, the LC(50) of M. anisopliae towards third instar larvae of H. armigera increased from the first sub-culture (0.17x10(4)) to (3.0x10(4)) for the 40th conidial transfers on potato dextrose agar and again decreased to 0.74x10(4) and 0.23x10(4) in the 40th (first in vivo) and the 40th (fifth in vivo) passage, respectively. Similar trends for LC(50) and LT(50) values were seen when sugarcane woolly aphid, Ceratovacuna lanigera Zehntner was used as a host. Significant variation in appressorium formation and cuticle-degrading enzyme production such as chitinase, chitin deacetylase, chitosanase and protease during subsequent sub-culturing and passage through H. armigera was observed. Though there was no effect on internal transcribed spacer (ITS) sequence pattern, interestingly, in randomly amplified polymorphic DNA (RAPD), significant differences in the band intensities and in the banding pattern for different sub-cultures of M. anisopliae were observed. As stable virulence towards the insect pest is desirable for commercialisation of a mycoinsecticide, such changes in virulence due to repeated in vitro transfer need to be monitored and minimised.</p>10aappressorium formation10achitin deacetylase10achitinase10aHelicoverpa armigera10ain vitro sub-culturing10aLipase10aMetarhizium anisopliae10aProtease1 aNahar, Pallavi, B.1 aKulkarni, Shuklangi, A.1 aKulye, Mahesh, S.1 aChavan, Santosh, B.1 aKulkarni, Girish1 aRajendran, Armugham1 aYadav, Priya, D.1 aShouche, Yogesh1 aDeshpande, Mukund, V.  uhttps://library.ncl.res.in/content/effect-repeated-vitro-sub-culturing-virulence-metarhizium-anisopliae-against-helicoverpa-002516nas a2200265   4500008004100000022001400041245012500055210006900180260007300249300001400322490000900336520153000345653001901875653002101894653003601915653001301951653002301964653003201987100002202019700001902041700001902060700002102079700002002100856013002120       2013                        eng d  a0304-416500aComplementation of intramolecular interactions for structural-functional stability of plant serine proteinase inhibitors0 aComplementation of intramolecular interactions for structuralfun  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcNOV  a5087-50940 v18303 a<p>Background: Plant protease inhibitors (PIs) constitute a diverse group of proteins capable of inhibiting proteases. Among Pls, serine Pls (SPIs) display stability and conformational restrictions of the reactive site loop by virtue of their compact size, and by the presence of disulfide bonds, hydrogen bonds, and other weak interactions. Scope of review: The significance of various intramolecular interactions contributing to protein folding mechanism and their role in overall stability and activity of SPIs is discussed here. Furthermore, we have reviewed the effect of variation or manipulation of these interactions on the activity/stability of SPIs. Major conclusions: The selective gain or loss of disulfide bond(s) in SPIs can be associated with their functional differentiation, which is likely to be compensated by non-covalent interactions (hydrogen bonding or electrostatic interactions). Thus, these intramolecular interactions are collectively responsible for the functional activity of SPIs, through the maintenance of scaffold framework, conformational rigidity and shape complementarities of reactive site loop. General significance: Structural insight of these interactions will provide an in-depth understanding of kinetic and thermodynamic parameters involved in the folding and stability mechanisms of SPIs. These features can be explored for engineering canonical SPIs for optimizing their overall stability and functionality for various applications. (C) 2013 Elsevier B.V. All rights reserved.</p>10aDisulfide bond10ahydrogen bonding10aIntramolecular weak interaction10aProtease10aReactive site loop10aSerine proteinase inhibitor1 aJoshi, Rakesh, S.1 aMishra, Manasi1 aSuresh, C., G.1 aGupta, Vidya, S.1 aGiri, Ashok, P.  uhttps://library.ncl.res.in/content/complementation-intramolecular-interactions-structural-functional-stability-plant-serine-000635nas a2200193   4500008004100000022001400041245007400055210006900129260000800198300000900206490000700215653001300222653002100235653001300256100002500269700002600294700001900320856010200339       2014                        eng d  a2053-273300aChickpea Kunitz Inhibitor: a mechanistic basis for trypsin inhibition0 aChickpea Kunitz Inhibitor a mechanistic basis for trypsin inhibi  cMAR  aC2710 v7010aChickpea10aKunitz Inhibitor10aProtease1 aBendre, Ameya, Dipak1 aRamasamy, Sureshkumar1 aSuresh, C., G.  uhttps://library.ncl.res.in/content/chickpea-kunitz-inhibitor-mechanistic-basis-trypsin-inhibition02351nas a2200277   4500008004100000022001400041245010100055210006900156260005700225300001000282490000700292520140100299653001901700653000801719653002201727653002101749653001301770653002201783100002101805700002301826700002101849700002601870700002101896700002501917856013101942       2015                        eng d  a1572-388700aAcid stability of the kinetically stable alkaline serine protease possessing polyproline II fold0 aAcid stability of the kinetically stable alkaline serine proteas  a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGERcFEB  a60-670 v343 a<p>The kinetically stable alkaline serine protease from Nocardiopsis sp.; NprotI, possessing polyproline II fold (PPII) was characterized for its pH stability using proteolytic assay, fluorescence and Circular Dichroism (CD) spectroscopy, and Differential Scanning Calorimetry (DSC). NprotI was found to be functionally stable when incubated at pH 1.0, even after 24 h, while after incubation at pH 10.0, drastic loss in the activity was observed. The enzyme showed enhanced activity after incubation at pH 1.0 and 3.0, at higher temperature (50-60 A degrees C). NprotI maintained the overall PPII fold in broad pH range as seen using far UV CD spectroscopy. The PPII fold of NprotI incubated at pH 1.0 remained fairly intact up to 70 A degrees C. Based on the isodichroic point and T-m values revealed by secondary structural transitions, different modes of thermal denaturation at pH 1.0, 5.0 and 10.0 were observed. DSC studies of NprotI incubated at acidic pH (pH 1.0-5.0) showed T-m values in the range of 74-76 A degrees C while significant decrease in T-m (63.8 A degrees C) was observed at pH 10.0. NprotI could be chemically denatured at pH 5.0 (stability pH) only with guanidine thiocynate. NprotI can be classified as type III protein among the three acid denatured states. Acid tolerant and thermostable NprotI can serve as a potential candidate for biotechnological applications.</p>10aAcid stability10aDSC10aKinetic stability10aPolyproline fold10aProtease10aThermal unfolding1 aRohamare, Sonali1 aJavdekar, Vaishali1 aDalal, Sayli, A.1 aNareddy, Pavan, Kumar1 aSwamy, Musti, J.1 aGaikwad, Sushama, M.  uhttps://library.ncl.res.in/content/acid-stability-kinetically-stable-alkaline-serine-protease-possessing-polyproline-ii-fold-002446nas a2200277   4500008004100000022001400041245012200055210006900177260009900246300001200345490000700357520142700364653001801791653002701809653002201836653001301858653002301871100002101894700002501915700001801940700002101958700001701979700001701996700002402013856013102037       2015                        eng d  a1359-511300aCloning, expression and in silico studies of a serine protease from a marine actinomycete (Nocardiopsis sp NCIM 5124)0 aCloning expression and in silico studies of a serine protease fr  aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcMAR  a378-3870 v503 a<p>A serine protease (N. protease), from Nocardiopsis sp., was cloned and expressed in Escherichia coli and investigated for its potential kinetic stability. Protein expression using two vectors, pET-22b (+) and pET-39b (+) was compared based on proper folding and soluble expression of the protein. pET-39b (+) was found to be a better vector for soluble expression of this protease containing disulfide bonds. In silico studies were also carried out for N. protease. Homology modeling suggested N. protease to be a member of PA clan of proteases. The phylogenetic analysis showed relatedness of N. protease to kinetically stable proteases. Molecular docking studies performed exhibited interaction of a peptide substrate with catalytic pocket of the enzyme. High temperature MD simulations were performed on N. protease to study its unfolding behavior and comparisons were made with alpha LP. A novel approach to study `cooperativity' of protein unfolding was undertaken, wherein `P' value analysis based on phi and psi values of the protein was performed. Data showed sharper P value transition for alpha LP when compared to N. protease thus indicating relatively less kinetic stability of N. protease. Present study holds significance as the non-streptomycete actinomycetes group is least explored and ensures industrially important enzymes with exceptional stabilities. (C) 2015 Elsevier Ltd. All rights reserved.</p>10aActinomycetes10aCloning and expression10aKinetic stability10aProtease10aThermal simulation1 aRohamare, Sonali1 aGaikwad, Sushama, M.1 aJones, Dafydd1 aBhavnani, Varsha1 aPal, Jayanta1 aSharma, Ranu1 aChatterjee, Prathit  uhttps://library.ncl.res.in/content/cloning-expression-and-silico-studies-serine-protease-marine-actinomycete-nocardiopsis-sp-002335nas a2200253   4500008004100000022001200041245015200053210006900205260000900274300001000283490000700293520141700300653002501717653001301742653002301755653003401778100002601812700002201838700002401860700002401884700002301908700002001931856013001951       2018                        eng d  a965174800aTripeptides derived from reactive centre loop of potato type II protease inhibitors preferentially inhibit midgut proteases of Helicoverpa armigera0 aTripeptides derived from reactive centre loop of potato type II   cAPR   a17-250 v953 a<p>Potato type II protease inhibitors (Pin-II PIs) impede the growth of lepidopteran insects by inhibiting serine protease-like enzymes in the larval gut. The three amino acid reactive centre loop (RCL) of these proteinaceous inhibitors is crucial for protease binding and is conserved across the Pin-II family. However, the molecular mechanism and inhibitory potential of the RCL tripeptides in isolation of the native protein has remained elusive. In this study, six peptides corresponding to the RCLs of the predominant Pin-II PIs were identified, synthesized and evaluated for in vitro and in vivo inhibitory activity against serine proteases of the polyphagous insect, Helicoverpa armigera. RCL peptides with sequences PRN, PRY and TRE were found to be potent inhibitors that adversely affected the growth and development of H. armigera. The binding mechanism and differential affinity of the RCL peptides with serine proteases was delineated by crystal structures of complexes of the RCL peptides with trypsin. Residues P1 and P2 of the inhibitors play a crucial role in the interaction and specificity of these inhibitors. Important features of RCL peptides like higher inhibition of insect proteases, enhanced efficacy at alkaline gut pH, longer retention and high stability in insect gut make them suitable molecules for the development of sustainable pest management strategies for crop protection.</p>10aHelicoverpa armigera10aProtease10aprotease inhibitor10aReactive centre loop peptides1 aSaikhedkar, Nidhi, S.1 aJoshi, Rakesh, S.1 aBhoite, Ashwini, S.1 aMohandasan, Radhika1 aYadav, Amit, Kumar1 aGiri, Ashok, P.  uhttps://library.ncl.res.in/content/tripeptides-derived-reactive-centre-loop-potato-type-ii-protease-inhibitors-preferentially