02417nas a2200217 4500008004100000022001400041245016500055210006900220260007300289300001400362490000900376520153500385653003201920653002701952653001901979653001101998653003402009100001602043700001402059856012602073 2006 eng d a0304-416500aBiochemical characterization of a low molecular weight aspartic protease inhibitor from thermo-tolerant bacillus licheniformis: kinetic interactions with pepsin0 aBiochemical characterization of a low molecular weight aspartic  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcDEC a1845-18560 v17603 a

The present article reports a low molecular weight aspartic protease inhibitor, API, from a newly isolated thermo-tolerant Bacillus licheniformis. The inhibitor was purified to homogeneity as shown by rp-HPLC and SDS-PAGE. API is found to be stable over a broad pH range of 2-11 and at temperature 90 degrees C for 2 1/2 h. It has a Mr (relative molecular mass) of 1363 Da as shown by MALDI-TOF spectra and 1358 Da as analyzed by SDS-PAGE The amino acid analysis of the peptide shows the presence of 12 amino acid residues having Mr of 1425Da. The secondary structure of API as analyzed by the CD spectra showed 7% alpha-helix, 49% beta-sheet and 44% aperiodic structure. The Kinetic studies of Pepsin-API interactions reveal that API is a slow-tight binding competitive inhibitor with the IC(50) and K(i) values 4.0nM and (3.83 nM - 5.31 nM) respectively. The overall inhibition constant K(i)* value is 0.107 +/- 0.015 nM. The progress curves are time-dependent and consistent with slow-tight binding inhibition: E+I reversible arrow (k(4), k(5)) EI reversible arrow (k(6), k(7)) EI*. Rate constant k(6) = 2.73 +/- 0.32 s(-1) reveals a fast isomerization of enzyme-inhibitor complex and very slow dissociation as proved by k(7)=0.068 +/- 0.009s(-1). The Rate constants from the intrinsic tryptophanyl fluorescence data is in agreement with those obtained from the kinetic analysis; therefore, the induced conformational changes were correlated to the isomerization of EI to EI*. (c) 2006 Elsevier B.V. Ail rights reserved.

10aAspartic protease inhibitor10aBacillus licheniformis10aenzyme kinetic10aPepsin10aSlow-tight binding inhibition1 aKumar, Ajit1 aRao, Mala uhttps://library.ncl.res.in/content/biochemical-characterization-low-molecular-weight-aspartic-protease-inhibitor-thermo-102062nas a2200265 4500008004100000022001400041245021000055210006900265260009500334300001200429490000800441520103600449653000601485653003701491653003301528653001301561653001101574653000901585653000901594653002401603653000901627100002101636700001401657856012501671 2006 eng d a0006-291X00aConformation and microenvironment of the active site of a low molecular weight 1,4-beta-D-glucan glucanohydrolase from an alkalothermophilic thermomonospora sp.: involvement of lysine and cysteine residues0 aConformation and microenvironment of the active site of a low mo a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcAUG a428-4320 v3473 a

Conformation and microenvironment at the active site of 1,4-beta-D-glucan glucanohydrolase was probed with fluorescent chemo-affinity labeling using o-phthalaldehyde. OPTA has been known to form a fluorescent isoindole derivative by cross-linking the proximal thiol and amino groups of cysteine and lysine. Modification of lysine of the enzyme by TNBS and of cysteine residue by PHMB abolished the ability of the enzyme to form an isoindole derivative with OPTA. Kinetic analysis of the TNBS and PHMB-modified enzyme suggested the presence of essential lysine and cysteine residues, respectively, at the active site of the enzyme. The substrate protection of the enzyme with carboxymethylcellulose (CMC) confirmed the involvement of lysine and cysteine residues in the active site of the enzyme. Multiple sequence alignment of peptides obtained by tryptic digestion of the enzyme showed cysteine is one of the conserved amino acids corroborating the chemical modification studies. (c) 2006 Elsevier Inc. All rights reserved.

10a110a4-beta-D-glucan glucanohydrolase10aactive site microenvironment10aCysteine10alysine10aOPTA10aPHMB10aThermomonospora sp.10aTNBS1 aJagtap, Sharmili1 aRao, Mala uhttps://library.ncl.res.in/content/conformation-and-microenvironment-active-site-low-molecular-weight-14-beta-d-glucan-001693nas a2200193 4500008004100000022001400041245010400055210006900159260007100228300001400299490000700313520095300320100001801273700002401291700002301315700001901338700001401357856012801371 2006 eng d a8756-793800aPreparation and characterization of urea-formaldehyde-pepsin bioconjugate: a new biocatalyst system0 aPreparation and characterization of ureaformaldehydepepsin bioco a1155 16TH ST, NW, WASHINGTON, DC 20036 USAbAMER CHEMICAL SOCcDEC a1585-15900 v223 a

This study describes the synthesis of urea formaldehyde (UF) microspheres by a dispersion polycondensation polymerization method. These microspheres with proper F/U molar ratio can provide highly reactive groups, capable of further condensation with the amino acid residues of enzyme/proteins. Presence of methylols groups in UF microspheres was confirmed by C-13 NMR study. Pepsin, a proteolytic enzyme, was immobilized on the UF microspheres to form bioconjugate system. As compared to the free enzyme in solution, the pepsin in the bioconjugate system exhibited significantly enhanced pH and temperature stability. The urea-formaldehydepepsin bioconjugate system also exhibited excellent proteolytic activity over eight successive reuse cycles with more than 50% of initial activity. A highlight of this new biocatalyst is the ease with which separation of this biocatalyst from the reaction medium may be achieved by mild centrifugation.

1 aVinod, V., P.1 aShinde, Sudhirkumar1 aD'Britto, Virginia1 aShukla, P., G.1 aRao, Mala uhttps://library.ncl.res.in/content/preparation-and-characterization-urea-formaldehyde-pepsin-bioconjugate-new-biocatalyst-002092nas a2200229 4500008004100000022001400041245010200055210006900157260006800226300001000294490000700304520127500311653002401586653001701610653001401627653001001641653002301651100002401674700003001698700001401728856012001742 2007 eng d a0006-359200aApplication of cellulases from an alkalothermophilic thermomonospora sp in biopolishing of denims0 aApplication of cellulases from an alkalothermophilic thermomonos a111 RIVER ST, HOBOKEN, NJ 07030 USAbJOHN WILEY & SONS INCcJAN a48-560 v963 a

Use of cellulase for denim washing is a standard eco-friendly technique to achieve desirable appearance and softness for cotton fabrics and denims. But enzymatic washing of denim till date involved acid cellulase (Trichoderma reesei) and neutral cellulase (Humicola isolens) the use of which has a drawback of backstaining of the indigo dye on to the fabric. Though it has been suggested that pH is a major factor in controlling backstaining there are no reports on use of cellulase under alkaline conditions for denim washing. In this study for the first time an alkali stable endoglucanase from alkalothermophilic Thermomonospora sp. (T-EG) has been used for denim biofinishing under alkaline conditions. T-EG is effective in removing hairiness with negligible weight loss and imparting softness to the fabric. Higher abrasive reactivity with lower backstaining was a preferred property for denim biofinishing exhibited by T-EG. The activities were comparable to acid and neutral cellulases that are being regularly used. The enzyme was also effective under non-buffering conditions which is an added advantage for use in textile industry. A probable mechanism of enzymatic finishing of cotton fabric has been represented based on the unique properties of T-EG.

10aalkaline conditions10abiopolishing10aCellulase10adenim10aThermomonospora sp1 aAnish, Ramakrishnan1 aRahman, Mohammad, Safikur1 aRao, Mala uhttps://library.ncl.res.in/content/application-cellulases-alkalothermophilic-thermomonospora-sp-biopolishing-denims01831nas a2200229 4500008004100000022001400041245006500055210006500120260009500185300001400280490000600294520104900300653001701349653001701366653002301383100002101406700002101427700001401448700002101462700001901483856009901502 2007 eng d a1533-488000aBacterial enzyme mediated biosynthesis of gold nanoparticles0 aBacterial enzyme mediated biosynthesis of gold nanoparticles a25650 NORTH LEWIS WAY, STEVENSON RANCH, CA 91381-1439 USAbAMER SCIENTIFIC PUBLISHERScDEC a4369-43770 v73 a

Development of synthesis methods for anisotropic metal nanoparticles is of considerable interest due to their remarkable optoelectronic properties. Various shapes ranging from rods to cubes to tetrapods and prisms may be obtained by chemical methods. Here we show that anisotropic gold nanoparticles can be synthesized biologically by the bacterium Actinobacter spp. when challenged with gold chloride in the presence of Bovine serum albumin (BSA). We also observed that synthesis of gold nanoparticles occur with simultaneous induction of the protease enzyme secreted by the bacterium in the presence of BSA. The presence of BSA helps to enhance the rate of gold nanoparticles biosynthesis and may also impart some shape control. Controlling simple experimental conditions like incubation temperature and presence or absence of oxygen have drastic effect on the reaction rate and the morphology of the particles. Various assay experiments show that the presence of enzyme protease can act as a reducing as well as shape directing agent.

10aBiosynthesis10abovine serum10agold nanoparticles1 aBharde, Atul, A.1 aKulkarni, Aarohi1 aRao, Mala1 aPrabhune, Asmita1 aSastry, Murali uhttps://library.ncl.res.in/content/bacterial-enzyme-mediated-biosynthesis-gold-nanoparticles-002040nas a2200241 4500008004100000022001400041245015900055210006900214260010300283300001400386490000700400520107700407653003401484653001301518653001701531653002101548653002301569653002401592653001301616100002401629700001401653856013101667 2007 eng d a0300-908400aBiochemical characterization of a novel beta-1-3, 1-4 glucan 4-glucanohydrolase from thermomonospora sp having a single active site for lichenan and xylan0 aBiochemical characterization of a novel beta13 14 glucan 4glucan a23 RUE LINOIS, 75724 PARIS, FRANCEbELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIERcDEC a1489-14970 v893 a

A bifunctional high molecular weight (Mr, 64,500 Da) beta-1-3, 1-4 glucan 4-glucanohydrolase was purified to homogeneity from Thermomonospora sp., exhibiting activity towards lichenan and xylan. A kinetic method was used to analyze the active site that hydrolyzes lichenan and xylan. The experimental data was in agreement with the theoretical values calculated for a single active site. Probing the conformation and microenvironment at active site of the enzyme by fluorescent chemo-affinity label, OPTA resulted in the formation of an isoindole derivative with complete inactivation of the enzyme to hydrolyse both lichenan and xylan confirmed the results of kinetic method. OPTA forms an isoindole derivative by cross-linking the proximal thiol and amino groups. The modification of cysteine and lysine residues by DTNB and TNBS respectively abolished the ability of the enzyme to form an isoindole derivative with OPTA, indicating the participation of cysteine and lysine in the formation of isoindole complex. (C) 2007 Elsevier Masson SAS. All rights reserved.

10a1-4 glucan 4-glucanohydrolase10abeta-1-310abifunctional10aO-phthaialdehyde10asingle active site10aThermomonospora sp.10aXylanase1 aAnish, Ramakrishnan1 aRao, Mala uhttps://library.ncl.res.in/content/biochemical-characterization-novel-beta-1-3-1-4-glucan-4-glucanohydrolase-thermomonospora-002197nas a2200217 4500008004100000022001400041245017500055210006900230260007900299300001200378490000900390520129600399653001501695653002401710653001901734653003601753653003201789100002101821700001401842856012301856 2007 eng d a0096-300300aBiochemical characterization of an aspartic protease from vigna radiata: kinetic interactions with the classical inhibitor pepstatin implicating a tight binding mechanism0 aBiochemical characterization of an aspartic protease from vigna  a360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USAbELSEVIER SCIENCE INCcMAY a619-6270 v17743 a

{Aspartic proteases are the focus of recent research interest in understanding the physiological importance of this class of enzymes in plants. This is the first report of an aspartic protease from the seeds of Vigna radiata. The aspartic protease was purified to homogeneity by fractional ammonium sulfate precipitation and pepstatin-A agarose affinity column. It was found to have a molecular weight of 67,406 Da by gel filtration chromatography. SDS-PAGE analysis revealed the presence of a heterodimer with subunits of molecular weights of 44,024 and 23,349 Da respectively. The enzyme was pH stable with the amino acid analysis confirming the molecular weight of the protein. The substrate cleavage site as analyzed by using the synthetic substrate was found to be the Phe-Tyr bond. The kinetic interactions of the enzyme were studied with the universal inhibitor, pepstatin A. This is the first report on the interactions of a plant aspartic protease with pepstatin-A, an inhibitor from a microbial source. A competitive one-step mechanism of binding is observed. The progress curves are time-dependent and consistent with tight binding inhibition. The K(i) value of the reversible complex of pepstatin with the enzyme was 0.87 mu M whereas the overall inhibition constant K(i)*

10aant colony10aglobal optimization10ametaheuristics10amultimodal continuous functions10aparticle swarm optimization1 aKulkarni, Aarohi1 aRao, Mala uhttps://library.ncl.res.in/content/biochemical-characterization-aspartic-protease-vigna-radiata-kinetic-interactions-001071nas a2200241 4500008004100000022001400041245009200055210006900147260007100216300001200287490000700299520024200306653002100548653000900569653002400578653001300602100002400615700001900639700002100658700001600679700001400695856012000709 2007 eng d a1525-779700aEncapsulation of endoglucanase using a biopolymer Gum arabic for its controlled release0 aEncapsulation of endoglucanase using a biopolymer Gum arabic for a1155 16TH ST, NW, WASHINGTON, DC 20036 USAbAMER CHEMICAL SOCcJAN a368-3720 v983 a

{Gum Arabic, a biodegradable natural polymer was used as a matrix to encapsulate endoglucanase from Thermomonospora sp. The modified enzyme retained complete biocatalytic activity and exhibited a shift in the optimum temperature [

10aEnzyme catalysis10aOPTA10aThermomonospora sp.10aXylanase1 aRamakrishnan, Anish1 aPandit, Nilesh1 aBadgujar, Mahesh1 aBhaskar, C.1 aRao, Mala uhttps://library.ncl.res.in/content/encapsulation-endoglucanase-using-biopolymer-gum-arabic-its-controlled-release-001496nas a2200217 4500008004100000022001400041245011700055210006900172260007900241300001000320490000800330520068300338653002101021653000901042653002401051653001301075100002501088700001901113700001401132856013201146 2007 eng d a1431-673000aGlycine-assisted enhancement of 1,4-beta-D-xylan xylanohydrolase activity at alkaline pH with a pH optimum shift0 aGlycineassisted enhancement of 14betaDxylan xylanohydrolase acti aGENTHINER STRASSE 13, D-10785 BERLIN, GERMANYbWALTER DE GRUYTER GMBHcJAN a61-650 v3883 a

This is the first report describing the enhancement of xylanase activity by the neutral amino acid glycine. Xylanase activity is increased seven-fold at alkaline pH in the presence of glycine and its pH optimum is shifted from pH 7 to 8 without using any protein engineering techniques. Analysis of the steady-state kinetics revealed that glycine in the reaction mixture increases the K-m and k(cat) values of the enzyme. Chemoaffinity labeling and studies using glycine esters indicate an involvement of the carboxylate ion of glycine in enhancing xylanase catalytic activity. A novel possible mechanism for the glycine-assisted catalytic action of xylanase is proposed.

10aEnzyme catalysis10aOPTA10aThermomonospora sp.10aXylanase1 aVathipadiekal, Vinod1 aVerma, Anamika1 aRao, Mala uhttps://library.ncl.res.in/content/glycine-assisted-enhancement-14-beta-d-xylan-xylanohydrolase-activity-alkaline-ph-ph-optimum01499nas a2200205 4500008004100000022001400041245013100055210006900186260007300255300001200328490000700340520067200347653003401019653002801053653002701081100002101108700002501129700001401154856012501168 2008 eng d a0141-813000apH Induced structural alterations in an aspartic protease from Vigna radiata indicating an alkali induced molten globule state0 apH Induced structural alterations in an aspartic protease from V aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcNOV a373-3760 v433 a

pH-dependent transitions in secondary and tertiary structure are described for a plant aspartic protease from Vigna radiata. The enzyme was pH stable with pH optima of 3.0. The Lineweaver Burk analysis at various pH yielded pK(a) values of 3.3 and 4.29 indicating acidic amino acids at the active site of the enzyme. The structural changes exemplified compact secondary structure collapsed tertiary structure and exposure of hydrophobic patches at pH 10. The changes at pH 10 are typical of a molten globule state. This alkali induced molten globule is novel since acid induced molten globule state is more reported. (C) 2008 Elsevier B.V. All rights reserved.

10aAlkali induced molten globule10aPlant aspartic protease10aStructural transitions1 aKulkarni, Aarohi1 aGaikwad, Sushama, M.1 aRao, Mala uhttps://library.ncl.res.in/content/ph-induced-structural-alterations-aspartic-protease-vigna-radiata-indicating-alkali-002543nas a2200205 4500008004100000022001400041245015800055210006900213260007900282300001400361490000700375520170700382653002202089653002002111653002402131653001802155100002102173700001402194856012902208 2009 eng d a0196-978100aDifferential elicitation of an aspartic protease inhibitor: regulation of endogenous protease and initial events in germination in seeds of vigna radiata0 aDifferential elicitation of an aspartic protease inhibitor regul a360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USAbELSEVIER SCIENCE INCcDEC a2118-21260 v303 a

Plant aspartic proteases are of recent origin with their physiological significance in crucial processes emerging. Reports on the significance of aspartic protease inhibitors and their endogenous proteases in seeds of plants are scanty. This paper reports the purification of an aspartic protease inhibitor from the seeds of Vigna radiata, its control of the endogenous aspartic protease and their subsequent role in the early germination events. The role of the aspartic protease inhibitor and the enzyme in initial stages of germination of V. radiata has been tracked by differential timed expression and germination assays. The expression pattern revealed maximum expression of the inhibitor in the dormant seeds while the enzyme was predominant in the germinating seeds. Their expression patterns and interactions indicate their significance in initiation of germination. The expression of other classes of proteases was monitored during germination and a model predicting the events occurring during proteolysis of the storage protein in germination is hypothesized. The inhibitor was a linear, hydrophobic, pH stable and thermostable peptide with molecular weight of 1660 Da. The purified inhibitor showed a pI of 4.36 with the sequence as AEIYN KDGNK LDLYG. The inhibitor was found to be stable in a broad range of pH from 2 to 10 with an optimum of 3.0. The half-life of VrAPI at 100 degrees C was 30 min whereas the maximum activity was observed at 37 degrees C. The initial kinetic analysis of the inhibitor against the enclogenous protease showed an IC50 value of 11 nM while the value of the inhibition rate constant K-i was 34 x 10(-9) M. (C) 2009 Elsevier Inc. All rights reserved.

10aAspartic protease10aPlant proteases10aProtease inhibitors10aVigna radiata1 aKulkarni, Aarohi1 aRao, Mala uhttps://library.ncl.res.in/content/differential-elicitation-aspartic-protease-inhibitor-regulation-endogenous-protease-and-002180nas a2200205 4500008004100000022001400041245018700055210006900242260007500311300001200386490000700398520133500405653000801740653001401748653002801762653002001790100002101810700001401831856012901845 2009 eng d a1053-050900aFluorescence study on interactions of alpha-crystallin with the molten globule state of 1, 4-beta-D-glucan glucanohydrolase from thermomonospora sp induced by guanidine hydrochloride0 aFluorescence study on interactions of alphacrystallin with the m a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGER/PLENUM PUBLISHERScNOV a967-9730 v193 a

In this paper, the interaction between alpha- crystallin and molten globule structure of 1,4-beta-D-Glucan Glucohydrolase (TSC) from an alkalothermophilic Thermomonospora sp. was investigated mainly by fluorescence quenching spectra, circular dichroism and three dimensional fluorescence spectra under simulative physiological conditions. Denaturation studies using GdnCl indicated that TSC folds through a partially folded state that resembles molten globule at 1.8 M GdnCl. The chaperone activity of alpha- crystallin was employed to study refolding of TSC. Here we studied the refolding of GdnCl denatured TSC from its molten globule state (TSC-m complex) in the presence and absence of alpha-crystallin to elucidate the molecular mechanism of chaperone-mediated in vitro folding. Our results, based on intrinsic tryptophan fluorescence and ANS binding studies, suggest that alpha-crystallin formed a complex with a putative intermediate molten globule - like intermediate in the refolding pathway of TSC. Reconstitution of the active TSC was observed on cooling the alpha-crystallin aEuro cent TSC -m complex to 4A degrees C. Addition of alpha-crystallin to the molten globule - like intermediate of TSC (TSC-m complex) complex initiated the refolding of TSC with 69 % recovery of the biological activity of the enzyme.

10aANS10aCellulase10aGuanidine hydrochloride10aProtein folding1 aJagtap, Sharmili1 aRao, Mala uhttps://library.ncl.res.in/content/fluorescence-study-interactions-alpha-crystallin-molten-globule-state-1-4-beta-d-glucan-001877nas a2200241 4500008004100000022001400041245012800055210006900183260009900252300001400351490000800365520092800373653002601301653002501327653002601352653002601378653002201404100002001426700002301446700002501469700001401494856012701508 2009 eng d a0960-852400aHydrolysis of cellulose derived from steam exploded bagasse by penicillium cellulases: comparison with commercial cellulase0 aHydrolysis of cellulose derived from steam exploded bagasse by p aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcDEC a6679-66810 v1003 a

A complete cellulase from Penicillium pinophilum was evaluated for the hydrolysis of a-cellulose derived from steam exploded sugarcane bagasse and other cellulosic substrates. alpha-Cellulose at 1% substrate concentration was completely hydrolyzed by Penicillium cellulase within 3 h wherein at 10% the hydrolysis was 100% within 24 h with an enzyme loading of 10 FPU/g. The hydrolysate yielded glucose as major end product as analyzed by HPLC. Under similar conditions, hydrolysis of Sigmacell (microcrystalline cellulose), CP-123 (pulverized cellulose powder) and ball milled Solka Floc were 42%, 56% and 52%, respectively. Further the hydrolysis performance of Penicillium sp. cellulase is compared with Trichoderma reesei cellulase (Accellerase (TM) 1000) from Genencore. The kinetics of hydrolysis with respect to enzyme and substrate concentration will be presented. (C) 2009 Elsevier Ltd. All rights reserved.

10aAccellerase (TM) 100010aEnzymatic hydrolysis10aHigh beta-glucosidase10aPenicillium cellulase10aSugarcane bagasse1 aSingh, Rajkumar1 aVarma, Anjanikumar1 aLaxman, Ryali, Seeta1 aRao, Mala uhttps://library.ncl.res.in/content/hydrolysis-cellulose-derived-steam-exploded-bagasse-penicillium-cellulases-comparison-001982nas a2200241 4500008004100000022001400041245016500055210006900220260009900289300001400388490000800402520103100410653001801441653001201459653001801471653002601489653002501515100001801540700001801558700002101576700001401597856012901611 2010 eng d a0960-852400aBiocatalytic approach for the utilization of hemicellulose for ethanol production from agricultural residue using thermostable xylanase and thermotolerant yeast0 aBiocatalytic approach for the utilization of hemicellulose for e aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcJUL a5366-53730 v1013 a

A hydrolysis of 62% and 50% for OSX (Oat spelt xylan) and WBH (Wheat bran hemicellulose) were obtained in 36 h and 48 h using Accellerase(TM) 1000 at 50 degrees C wherein thermostable xylanase from alkalothermophilic Thermomonospora sp. yielded 67% (OSX) in 3 h and 58% (WBH) in 24 h at 60 degrees C, favouring a reduction in process time and enzyme dosage. The rate of hydrolysis with thermostable xylanase was increased by 20% with the addition of nonionic surfactant tween 80 or biosurfactant sophorolipid. The simultaneous saccharification and fermentation (SSF) of OSX and WBH using thermostable xylanase and D. hansenii in batch cultures produced 9.1 g/L and 9.5 g/L of ethanol, respectively and had a shorter overall process time than the separate hydrolysis and fermentation (SHF). The immobilized yeast cells in Ca-alginate matrix produced ethanol with a yield of 0.46 g/g from hemicellulosic hydrolysates and were reused six times with 100% fermentation efficiency. (C) 2010 Elsevier Ltd. All rights reserved.

10aBiosurfactant10aEthanol10aHemicellulose10aThermostable xylanase10aThermotolerant yeast1 aMenon, Vishnu1 aPrakash, Gyan1 aPrabhune, Asmita1 aRao, Mala uhttps://library.ncl.res.in/content/biocatalytic-approach-utilization-hemicellulose-ethanol-production-agricultural-residue-002367nas a2200241 4500008004100000022001400041245016600055210006900221260007300290300001200363490000800375520144300383653001201826653002201838653001501860653002701875653002501902653001801927100001801945700001801963700001401981856013001995 2010 eng d a0168-165600aEnzymatic hydrolysis and ethanol production using xyloglucanase and debaromyces hansenii from tamarind kernel powder: galactoxyloglucan predominant hemicellulose0 aEnzymatic hydrolysis and ethanol production using xyloglucanase  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcAUG a233-2390 v1483 a

The hydrolysis and ethanol production from tamarind kernel powder (TKP), a rich source of galactoxyloglucan (GXG) was investigated for the first time using xyloglucanase and thermotolerant Debaromyces hansenii. The acid hydrolysis of TKP with 2N H(2)SO(4) at 120 degrees C for 30 min yielded an overall saccharification of 94% based on the total available carbohydrate content and further fermentation at 40 degrees C with thermotolerant D hansenii produced an ethanol yield of 0.35 g/g. A maximum hydrolysis of 55 and 78% for GXG was obtained in 48 h at 50 degrees C using Thermomonospora xyloglucanase (TXy) and accellerase (TM) 1000, respectively. The synergistic effect of beta-galactosidase and xyloglucanase was demonstrated by the exogenous addition of beta-galactosidase to TXy which improved the overall hydrolysis of GXG by 30%. The rate of hydrolysis of GXG with TXy and accellerase was increased by 15-20% in the presence of chemical surfactants (tween 80 and toluene) or protein additive (BSA). The fermentation of enzymatic hydrolysates of GXG by TXy and accellerase with free cells at 40 degrees C produced an ethanol yield of 0.39 and 0.41 g/g whereas with immobilized cells produced 0.45 and 0.43 g/g, respectively, with a theoretical conversion efficiencies of 78-88%. The immobilized yeast cells were reused six times at 40 degrees C with 100% fermentation efficiency. (C) 2010 Elsevier B.V. All rights reserved.

10aEthanol10aGalactoxyloglucan10aHydrolysis10aTamarind kernel powder10aThermotolerant yeast10aXyloglucanase1 aMenon, Vishnu1 aPrakash, Gyan1 aRao, Mala uhttps://library.ncl.res.in/content/enzymatic-hydrolysis-and-ethanol-production-using-xyloglucanase-and-debaromyces-hansenii-002308nas a2200241 4500008004100000022001400041245015900055210006900214260007900283300001000362490000700372520137500379653002001754653001901774653001401793653001401807653001901821100002501840700003401865700002301899700001401922856013001936 2010 eng d a0196-978100aMolecular cloning, over expression, and activity studies of a peptidic HIV-1 protease inhibitor: designed synthetic gene to functional recombinant peptide0 aMolecular cloning over expression and activity studies of a pept a360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USAbELSEVIER SCIENCE INCcJAN a16-210 v313 a

The aspartic protease inhibitor (ATBI) purified from a Bacillus sp. is a potent inhibitor of several proteases including recombinant HIV-1 protease, pepsin, and fungal aspartic protease. In this study, we report the cloning, and over expression of a synthetic gene coding for ATBI in Escherichia coli and establish a purification Protocol. The ATBI molecule consists of eleven amino acids and is peptidic in nature. We used the peptide sequence data of ATBI to synthesize complementary oligonucleotides, which were annealed and subsequently cloned in-frame with the gene for glutathione-S-transferase (GST). The expression of the resulting fusion protein was induced in E. coli BL21-A1 cells using arabinose. The recombinant peptide was Purified using a reduced glutathione column, and cleaved with Factor Xa to remove the GST tag. The resultant product was further purified to homogeneity using RP-HPLC. Mass spectroscopy analysis revealed that the purified peptide had a molecular weight of 1186 Da which matches the theoretical molecular weight of the amino acids present in the synthetic gene. The recombinant peptide was found to be active in vitro against HIV-1 protease, pepsin, and fungal aspartic protease. The protocol described in this study may be used to clone pharmaceutically important peptide molecules. (C) 2009 Elsevier Inc. All rights reserved.

10aGateway cloning10aHIV-1 protease10ainhibitor10aProteases10aSynthetic gene1 aVathipadiekal, Vinod1 aUmasankar, Perunthottathu, K.1 aPatole, Milind, S.1 aRao, Mala uhttps://library.ncl.res.in/content/molecular-cloning-over-expression-and-activity-studies-peptidic-hiv-1-protease-inhibitor-002979nas a2200217 4500008004100000022001400041245015900055210006900214260007300283300001200356490000900368520210000377653003202477653002702509653001602536653002002552653003402572100001602606700001402622856012502636 2010 eng d a0304-416500aNovel bifunctional peptidic aspartic protease inhibitor inhibits chitinase A from Serratia marcescens: kinetic analysis of inhibition and binding affinity0 aNovel bifunctional peptidic aspartic protease inhibitor inhibits aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcMAY a526-5360 v18003 a

Background: Chitinase inhibitors have chemotherapeutic potential as fungicides, pesticides and antiasthmatics. The majority of chitinase inhibitors reported are natural products like argifin, argifin linear fragments, argadin, allosamidin and disulfide-cyclized peptides. Here, we report a novel peptidic inhibitor API (Aspartic Protease Inhibitor), isolated from Bacillus licheniforrnis that inhibits chitinase A (ChiA) from Serratia marcescens. Methods: The binding affinity of API with ChiA and type of inhibition was determined by the inhibition kinetics assays. Fluorescence and CD spectroscopic analysis and chemical modification of API with different affinity reagents elucidated the mechanism of binding of API with ChiA. Results and conclusions: The peptide has an amino acid sequence N-Ile(1)-Cys(2)-Glu(3)-Ala(4)-Glu(5)-His(6)-Lys(7)-Trp(8)-Gly(9)-Asp(10) -Tyr(11)-Leu(12)-Asp(13)-C. The ChiA-API kinetic interactions reveal noncompetitive, irreversible and tight binding nature of API with I-50= 600 nM and K-i = 510 nM in the presence of chromogenic substrate pnitrophenyl-N,N'-diacetyl-beta-chitobioside[p-NP-(GIcNAc)(2)]. The inhibition progress curves show a two-step slow tight binding inhibition mechanism with the rate constant k(5) = 8.7 +/- 1 x 10(-3) s(-1) and k(6) = 7.3 +/- 0.6 x 10(-5) s(-1). CD-spectra and tryptophanyl fluorescence analysis of ChiA incubated with increasing API concentrations confirms conformational changes in enzyme structure which may be due to irreversible denaturation of enzyme upon binding of API. Chemical modifications by WRK abolished the anti-chitinase activity of API and revealed the involvement of carboxyl groups in the enzyme inactivation. Abolished isoindole fluorescence of OPTA-labeled ChiA demonstrates the irreversible denaturation of ChiA upon incubation with API for prolonged time and distortion of active site of the enzyme. General significance: The data provide useful information that could lead to the generation of drug-like, natural product-based chitinase inhibitors. (C) 2010 Elsevier B.V. All rights reserved.

10aAspartic protease inhibitor10aBacillus licheniformis10aChitinase A10aEnzyme kinetics10aSlow tight binding inhibition1 aKumar, Ajit1 aRao, Mala uhttps://library.ncl.res.in/content/novel-bifunctional-peptidic-aspartic-protease-inhibitor-inhibits-chitinase-serratia-001496nas a2200229 4500008004100000022001400041245010300055210006900158260010200227300001400329490000600343520060500349653002400954653001800978653003300996653002901029653001301058100001401071700002301085700002601108856013201134 2010 eng d a1930-212600aProduction of single cell protein, essential amino acids, and xylanase by penicillium janthinellum0 aProduction of single cell protein essential amino acids and xyla aCAMPUS BOX 8005, RALEIGH, NC 27695-8005 USAbNORTH CAROLINA STATE UNIV DEPT WOOD & PAPER SCIcOCT a2470-24770 v53 a

Microbial biomass having 46% crude protein content and enriched with essential amino acids as well as extracellular xylanase activity (100-150 IU/ml) was produced by an efficient fungal strain, Penicillium janthinellum (NCIM St-F-3b). Optimization studies for maximum xylanase and biomass production showed that the fungus required a simple medium containing bagasse hemicellulose as carbon source and ammonium sulphate as the nitrogen source. Therefore bagasse, which is a waste product of the sugar industry, can be efficiently used in microbioal biomass protein preparation for animal feed.

10aBagasse hydrolysate10aCrude protein10aMicrobial biomass production10aPenicillium janthinellum10aXylanase1 aRao, Mala1 aVarma, Anjanikumar1 aDeshmukh, Sumedha, S. uhttps://library.ncl.res.in/content/production-single-cell-protein-essential-amino-acids-and-xylanase-penicillium-janthinellum-002048nas a2200241 4500008004100000022001400041245014500055210006900200260007300269300001200342490000700354520117600361653001201537653001501549653001301564653001401577653001401591653002501605100001801630700001901648700001401667856012501681 2011 eng d a0378-382000aBioethanol production from renewable polymer lichenan using lichenase from an alkalothermophilic thermomonospora sp and thermotolerant yeast0 aBioethanol production from renewable polymer lichenan using lich aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcMAR a401-4060 v923 a

Biomass feedstocks available decentrally will be more commodious for localized biorefinery approach than the exhaustive large scale and centralized plants driven by cost intensive technology. Lichen is present in a wide range of habitats in a distributed manner. A maximum hydrolysis of 73%-76% for lichenan from Cetraria islandica, Usnea barbata and Parmelia sp. were obtained in 24 h using lichenase from an alkalothermophilic Thermomonospora sp. wherein the hydrolysis was 100% with commercial enzyme Accellerase (TM) 1000. The synergistic role of beta-glucosidase in lichenan hydrolysis was demonstrated by the exogenous addition of beta-glucosidase to Thermomonospora lichenase which resulted in complete hydrolysis. The hydrolysates of lichenan obtained using Accellerase or a cocktail of Thermomonospora lichenase and beta-glucosidase when fermented with free cells of Saccharomyces at 40 degrees C produced an ethanol yield of 0.45 g/g-0.48 g/g with theoretical conversion efficiencies of 93%-96%. The Ca-alginate immobilized yeast cells were reused eight times at 40 degrees C with 100% fermentation efficiency. (C) 2010 Elsevier B.V. All rights reserved.

10aEthanol10aHydrolysis10aLichenan10aLichenase10aSynergism10aThermotolerant yeast1 aMenon, Vishnu1 aDivate, Rupesh1 aRao, Mala uhttps://library.ncl.res.in/content/bioethanol-production-renewable-polymer-lichenan-using-lichenase-alkalothermophilic-002101nas a2200253 4500008004100000022001400041245015000055210006900205260009900274300001400373490000800387520111000395653001901505653003601524653004101560653001201601653001101613100001801624700002301642700002101665700002001686700001401706856012701720 2011 eng d a0960-852400aMicrobial production of xylitol from D-xylose and sugarcane bagasse hemicellulose using newly isolated thermotolerant yeast Debaryomyces hansenii0 aMicrobial production of xylitol from Dxylose and sugarcane bagas aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcFEB a3304-33080 v1023 a

A thermotolerant yeast capable of fermenting xylose to xylitol at 40 C was isolated and identified as a strain of Debaryomyces hansenii by ITS sequencing. This paper reports the production of xylitol from D-xylose and sugarcane bagasse hemicellulose by free and Ca-alginate immobilized cells of D. hansenii. The efficiency of free and immobilized cells were compared for xylitol production from D-xylose and hemicellulose in batch culture at 40 degrees C. The maximum xylitol produced by free cells was 68.6 g/L from 100 g/L of xylose, with a yield of 0.76 g/g and volumetric productivity 0.44 g/L/h. The yield of xylitol and volumetric productivity were 0.69 g/g and 0.28 g/L/h respectively from hemicellulosic hydrolysate of sugarcane bagasse after detoxification with activated charcoal and ion exchange resins. The Ca-alginate immobilized D. hansenii cells produced 73.8 g of xylitol from 100 g/L of xylose with a yield of 0.82 g/g and volumetric productivity of 0.46 g/L/h and were reused for five batches with steady bioconversion rates and yields. (C) 2010 Elsevier Ltd. All rights reserved.

10aImmobilization10aSugarcane bagasse hemicellulose10aThermotolerant Debaryomyces hansenii10axylitol10aXylose1 aPrakash, Gyan1 aVarma, Anjanikumar1 aPrabhune, Asmita1 aShouche, Yogesh1 aRao, Mala uhttps://library.ncl.res.in/content/microbial-production-xylitol-d-xylose-and-sugarcane-bagasse-hemicellulose-using-newly-001855nas a2200217 4500008004100000022001400041245009400055210006900149260009900218300001200317490000700329520104200336653002201378653002401400653001701424653002201441100001801463700001401481700002501495856011701520 2011 eng d a1359-511300aPurification and characterization of an alkaline protease by a new strain of Beauveria sp0 aPurification and characterization of an alkaline protease by a n aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcFEB a579-5850 v463 a

A fungal culture isolated from animal dung was identified as a new strain of Beauveria sp MTCC 5184 based on 18S rDNA and ITS nucleotide sequence homology. The fungal isolate secretes alkaline protease active at pH 9 and 50 degrees C. The alkaline protease from Beauveria sp (BAP) was purified to homogeneity with 10.2-folds increase in specific activity and 38.6% recovery. The molecular mass and isoelectric point of the protease were found to be 29 kDa and 9.3, respectively. The N-terminal sequence of the BAP showed only partial homology with subtilisin like proteases from other fungi. The enzyme was stable up to 40 degrees C and pH 3-11. The protease was inhibited by Cd(2+), Hg(2+) and Mn(2+). The activity was totally lost in the presence of 1 mM PMSF suggesting it to be a serine protease. The protease showed maximum activity with casein followed by haemoglobin and BSA. The purified protease is able to separate the endothelial cells and can be used in animal cell culture. (C) 2010 Elsevier Ltd. All rights reserved.

10aAlkaline Protease10aAnimal cell culture10aBeauveria sp10aSequence homology1 aShankar, Shiv1 aRao, Mala1 aLaxman, Ryali, Seeta uhttps://library.ncl.res.in/content/purification-and-characterization-alkaline-protease-new-strain-beauveria-sp-002277nas a2200229 4500008004100000022001400041245011800055210006900173260010700242300001200349490000700361520138000368653004301748653003401791653001701825653001701842653001801859100001601877700001801893700001401911856012201925 2012 eng d a1431-065100aBiochemical characterization of a novel thermostable xyloglucanase from an alkalothermophilic thermomonospora sp.0 aBiochemical characterization of a novel thermostable xyloglucana aCHIYODA FIRST BLDG EAST, 3-8-1 NISHI-KANDA, CHIYODA-KU, TOKYO, 101-0065, JAPANbSPRINGER JAPAN KKcJAN a135-1460 v163 a

Xyloglucanase from an extracellular culture filtrate of alkalothermophilic Thermomonospora sp. was purified to homogeneity with a molecular weight of 144 kDa as determined by SDS-PAGE and exhibited specificity towards xyloglucan with apparent K (m) of 1.67 mg/ml. The enzyme was active at a broad range of pH (5-8) and temperatures (40-80A degrees C). The optimum pH and temperature were 7 and 70A degrees C, respectively. The enzyme retained 100% activity at 50A degrees C for 60 h with half-lives of 14 h, 6 h and 7 min at 60, 70 and 80A degrees C, respectively. The kinetics of thermal denaturation revealed that the inactivation at 80A degrees C is due to unfolding of the enzyme as evidenced by the distinct red shift in the wavelength maximum of the fluorescence profile. Xyloglucanase activity was positively modulated in the presence of Zn2+, K+, cysteine, beta-mercaptoethanol and polyols. Thermostability was enhanced in the presence of additives (polyols and glycine) at 80A degrees C. A hydrolysis of 55% for galactoxyloglucan (GXG) from tamarind kernel powder (TKP) was obtained in 12 h at 60A degrees C and 6 h at 70A degrees C using thermostable xyloglucanases, favouring a reduction in process time and enzyme dosage. The enzyme was stable in the presence of commercial detergents (Ariel), indicating its potential as an additive to laundry detergents.

10aAlkalothermophilic Thermomonospora sp.10aBiotechnological applications10aPurification10aThermostable10aXyloglucanase1 aPol, Dipali1 aMenon, Vishnu1 aRao, Mala uhttps://library.ncl.res.in/content/biochemical-characterization-novel-thermostable-xyloglucanase-alkalothermophilic-001950nas a2200241 4500008004100000022001400041245010000055210006900155260007900224300001400303490000800317520106300325653001301388653002401401653002501425653002701450653002601477100002201503700002001525700002501545700001401570856012401584 2012 eng d a0273-228900aEffect of alkali pretreatment on the structural properties and enzymatic hydrolysis of corn cob0 aEffect of alkali pretreatment on the structural properties and e a999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USAbHUMANA PRESS INCcDEC a1806-18190 v1683 a

An effective alkali pretreatment which affects the structural properties of cellulose (corn cob) has been studied. The pretreatment of corn cob was carried out with different combinations of alkali at varying temperatures. The most effective pretreatment of corn cob was achieved with 1 % alkali at 50 A degrees C in 4 h. The crystallinity index (CrI) and specific surface area (SSA) of untreated corn cob was 39 % and 0.52 m(2)/g wherein after alkali pretreatment CrI decreased to 15 % and SSA increased to 3.32 m(2)/g. The fungal organism was identified as Penicillium pinophilum on the basis of ITS sequence. At 5 % substrate concentration using a complete cellulase from Penicillium pinophilum the hydrolysis of untreated corn cob with 5, 10 and 20 FPU/g enzyme loadings were 11 %, 13 % and 16 %, whereas after alkali treatment the hydrolysis increased to 78 %, 90 % and 100 %, respectively. Further hydrolytic potential of commercial cellulases viz. Accellerase (TM) 1,000, Palkofeel-30 and Palkocel-40 were investigated under similar conditions.

10aCorn cob10aCrystallinity index10aEnzymatic hydrolysis10aPenicillium pinophilum10aSpecific surface area1 aSahare, Padmavati1 aSingh, Rajkumar1 aLaxman, Ryali, Seeta1 aRao, Mala uhttps://library.ncl.res.in/content/effect-alkali-pretreatment-structural-properties-and-enzymatic-hydrolysis-corn-cob-001772nas a2200217 4500008004100000022001400041245014500055210006900200260009900269300001200368490000800380520088800388653002201276653002701298653002101325653002701346653001801373100001801391700001401409856013101423 2012 eng d a0960-852400aInhibition of xyloglucanase from an alkalothermophilic thermomonospora sp by a peptidic aspartic protease inhibitor from Penicillium sp VM240 aInhibition of xyloglucanase from an alkalothermophilic thermomon aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcNOV a390-3990 v1233 a

A bifunctional inhibitor from Penicilliurn sp VM24 causing inactivation of xyloglucanase from Thermomonospora sp and an aspartic protease from Aspergillus saitoi was identified. Steady state kinetics studies of xyloglucanase and the inhibitor revealed an irreversible, non-competitive, two-step inhibition mechanism with IC50 and K-i; values of 780 and 500 nM respectively. The interaction of o-phthalaldehyde (OPTA)-labeled xyloglucanase with the inhibitor revealed that the inhibitor binds to the active site of the enzyme. Far- and near-UV spectrophotometric analysis suggests that the conformational changes induced in xyloglucanase by the inhibitor may be due to irreversible denaturation of enzyme. The bifunctional inhibitor may have potential as a biocontrol agent for the protection of plants against phytopathogenic fungi. (C) 2012 Elsevier Ltd. All rights reserved.

10aAspartic protease10aBifunctional inhibitor10aBiocontrol agent10aInactivation mechanism10aXyloglucanase1 aMenon, Vishnu1 aRao, Mala uhttps://library.ncl.res.in/content/inhibition-xyloglucanase-alkalothermophilic-thermomonospora-sp-peptidic-aspartic-protease-001799nas a2200157 4500008004100000022001400041245012500055210006900180260011800249300001400367490000800381520108800389100001801477700001401495856013201509 2012 eng d a1350-087200aLow-molecular-mass aspartic protease inhibitor from a novel penicillium sp.: implications in combating fungal infections0 aLowmolecularmass aspartic protease inhibitor from a novel penici aMARLBOROUGH HOUSE, BASINGSTOKE RD, SPENCERS WOODS, READING RG7 1AG, BERKS, ENGLANDbSOC GENERAL MICROBIOLOGYcJUL a1897-19070 v1583 a

A low-molecular-mass aspartic protease inhibitor was isolated from a novel Penicillium sp. The inhibitor was purified to homogeneity, as shown by reversed-phase HPLC and SDS-PAGE. The M-r of the inhibitor was 1585 and the amino acid composition showed the presence of D, D, D, E, A, K, L, Y, H, I and W residues. The steady-state kinetic interactions of Aspergillus saitoi aspartic protease with the inhibitor revealed the reversible, competitive, time-dependent tight-binding nature of the inhibitor, with IC50 and K-i values of 1.8 and 0.85 mu M, respectively. Fluorescence spectroscopy and circular dichroism analysis showed that inactivation of the enzyme was due to binding of the inhibitor to the active site. The inhibitor was found to inhibit mycelial growth and spore germination of Aspergillus fumigatus and Aspergillus niger in vitro with MIC values of 1.65 and 0.30 mu g ml(-1), respectively. This study will potentially open the way towards the development of a tight-binding peptidic inhibitor against fungal aspartic proteases to combat human fungal infections.

1 aMenon, Vishnu1 aRao, Mala uhttps://library.ncl.res.in/content/low-molecular-mass-aspartic-protease-inhibitor-novel-penicillium-sp-implications-combating-002491nas a2200253 4500008004100000022001400041245010500055210006900160260007900229300001200308490000800320520154900328653002101877653002101898653003301919653001701952653002001969100002101989700002702010700002602037700001402063700002802077856013202105 2012 eng d a0273-228900aProduction, purification, and characterization of a beta-glucosidase of penicillium funiculosum NCL10 aProduction purification and characterization of a betaglucosidas a999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USAbHUMANA PRESS INCcJUL a959-9720 v1673 a

Penicillium funiculosum NCL1, a filamentous fungus, produced significantly higher levels of beta-glucosidase. The effect of initial pH, incubation temperature, and different carbon sources on extracellular beta-glucosidase production was studied in submerged fermentation. At 30 A degrees C with initial pH 5.0, enzyme production was increased by 48-fold upon induction with paper mill waste, as compared to commercial cellulose powder. In zymogram analysis, four isoforms of beta-glucosidases were observed with wheat bran whereas a minimum of one isoform was observed with other carbon sources. A major beta-glucosidase (Bgl3A) with the apparent molecular weight of similar to 120 kDa, induced by paper mill waste, was purified 19-fold to homogeneity, with a specific activity of 1,796 U/mg. Bgl3A was a monomeric glycoprotein with 29% of neutral carbohydrate content. It showed optimum activity at pH 4.0 and 5.0, optimum temperature at 60 A degrees C, and exhibited a half-life of 1 h at 60 A degrees C. K (m) of Bgl3A was found to be 0.057 mM with p-nitrophenyl beta-d-glucoside and V (max) was 1,920 U/mg. The purified enzyme exhibited glucose tolerance with a K (i) of 1.5 mM. Bgl3A readily hydrolyzed glucosides with beta-linkage. Bgl3A activity was enhanced (156%) by Zn2+ and was not affected by other metal cations and reagents. The supplementation of Bgl3A (5 U/mg) with Trichoderma reesei cellulase complex (5 FPU/mg) resulted in about 70% of enhanced glucose production, which emphasizes the industrial importance of Bgl3A.

10abeta-Glucosidase10aPaper mill waste10aPenicillium funiculosum NCL110aPurification10aThermostability1 aRamani, Gurusamy1 aMeera, Balasubramanian1 aVanitha, Chinnathambi1 aRao, Mala1 aGunasekaran, Paramasamy uhttps://library.ncl.res.in/content/production-purification-and-characterization-beta-glucosidase-penicillium-funiculosum-ncl1-002494nas a2200229 4500008004100000022001400041245010100055210006900156260011000225300001200335490000700347520160200354653002701956653002801983653001802011653003302029653001702062100001602079700002502095700001402120856013002134 2012 eng d a0301-120800aPurification and biochemical characterization of endoglucanase from Penicillium pinophilum MS 200 aPurification and biochemical characterization of endoglucanase f aDR K S KRISHNAN MARG, PUSA CAMPUS, NEW DELHI 110 012, INDIAbNATL INST SCIENCE COMMUNICATION-NISCAIRcJUN a189-1940 v493 a

Cellulases find increasing prominence in sustainable production of fuel and feedstock from lignocellulosic biomass. The purification and biochemical characterization of individual components of cellulase complex is important to understand the mechanism of their action for the solubilization of crystalline cellulose. In this study, an extra-cellular endoglucanase isolated from culture filtrate of Penicillium pinophilum MS 20 was purified to homogeneity by ammonium sulphate precipitation, ion-exchange chromatography and gel filtration. The purified endoglucanase (specific activity 69 U/mg) was a monomeric protein with molecular mass of 42 kDa, as determined by SDS-PAGE. The endoglucanase was active over a broad range of pH (4-7) with maximum activity at pH 5 and showed optimum temperature of 50 degrees C. It retained 100% activity at 50 degrees C for 6 h and half- lives of 4 h and 3 h at 60 degrees C and 70 degrees C, respectively. The kinetic constants for the endoglucanase determined with carboxymethyl cellulose as substrate were V-max of 72.5 U/mg and apparent K-m of 4.8 mg/ml. The enzyme also showed moderate activity towards H3PO4 swollen cellulose and p-nitrophenyl beta-D-glucoside, but no activity towards filter paper, Avicel and oat spelt xylan. The activity was positively modulated by 47, 32 and 25% in the presence of Co2+, Zn2+ and Mg2+, respectively to the reaction mixture. The wide pH stability (4-7) and temperature stability up to 50 degrees C of endoglucanase makes the enzyme suitable for use in cellulose saccharification at moderate temperature and pH.

10aBiochemical properties10aCarboxymethyl cellulose10aEndoglucanase10aPenicillium pinophilum MS 2010aPurification1 aPol, Dipali1 aLaxman, Ryali, Seeta1 aRao, Mala uhttps://library.ncl.res.in/content/purification-and-biochemical-characterization-endoglucanase-penicillium-pinophilum-ms-20-001448nas a2200193 4500008004100000022001400041245010500055210006900160260007300229300001200302490000700314520068200321653003201003653002501035653003401060100001801094700001401112856012801126 2012 eng d a0141-813000aSlow-tight binding inhibition of pepsin by an aspartic protease inhibitor from streptomyces sp MBR040 aSlowtight binding inhibition of pepsin by an aspartic protease i aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcJUL a165-1740 v513 a

The present article reports a low molecular weight aspartic protease inhibitor from a Streptomyces sp. MBR04 exhibiting a two-step inhibition mechanism against pepsin. The kinetic interactions revealed a reversible, competitive, slow-tight binding inhibition with an IC50 and K-i values of 4.5 nM and 4 nM respectively. The conformational changes induced upon inhibitor binding to pepsin was monitored by far and near UV analysis, demonstrated that the inhibitor binds to the active site and causes inactivation. Chemical modification of the inhibitor with WRK and TNBS abolished the antiproteolytic activity of the inhibitor. (c) 2012 Elsevier B.V. All rights reserved.

10aAspartic protease inhibitor10aLow molecular weight10aSlow-tight binding inhibition1 aMenon, Vishnu1 aRao, Mala uhttps://library.ncl.res.in/content/slow-tight-binding-inhibition-pepsin-aspartic-protease-inhibitor-streptomyces-sp-mbr04-002760nas a2200229 4500008004100000022001400041245009800055210006900153260010600222300001200328490000700340520187700347653003602224653002502260653001902285653001802304653002602322653002502348100001802373700001402391856012502405 2012 eng d a0360-128500aTrends in bioconversion of lignocellulose: biofuels, platform chemicals & biorefinery concept0 aTrends in bioconversion of lignocellulose biofuels platform chem aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcAUG a522-5500 v383 a

Bioconversion of renewable lignocellulosic biomass to biofuel and value added products are globally gaining significant prominence. Market forces demonstrate a drive towards products benign to natural environment increasing the importance of renewable materials. The development of second generation bioethanol from lignocellulosic biomass serves many advantages from both energy and environmental point of views. Biomass an inexpensive feedstock considered sustainable and renewable, is an option with the potential to replace a wide diversity of fossil based products within the energy sector; heat, power, fuels, materials and chemicals. Lignocellulose is a major structural component of woody and non-woody plants and consists of cellulose, hemicellulose and lignin. The effective utilization of all the three components would play a significant role in the economic viability of cellulosic ethanol. Biomass conversion process involves five major steps, choice of suitable biomass, effective pretreatment, production of saccharolytic enzymes-cellulases and hemicellulases, fermentation of hexoses and pentoses and downstream processing. Within the context of production of fuels from biomass, pretreatment has come to denote processes by which cellulosic biomass is made amenable to the action of hydrolytic enzymes. The limited effectiveness of current enzymatic process on lignocellulose is thought to be due to the relative difficulties in pretreating the feedstocks. The present review is a comprehensive state of the art describing the advancement in recent pretreaments, metabolic engineering approaches with special emphasis on the latest developments in consolidated biomass processing, current global scenario of bioethanol pilot plants and biorefinery concept for the production of biofuels and bioproducts. (C) 2012 Elsevier Ltd. All rights reserved.

10aConsolidated biomass processing10aLifecycle assessment10aLignocellulose10aPre-treatment10aSaccharifying enzymes10aValue-added products1 aMenon, Vishnu1 aRao, Mala uhttps://library.ncl.res.in/content/trends-bioconversion-lignocellulose-biofuels-platform-chemicals-biorefinery-concept-002381nas a2200265 4500008004100000022001400041245014900055210006900204260009900273300001000372490000700382520140800389653000601797653000801803653003301811653002301844653001201867653001501879653001401894653002501908100001901933700001801952700001401970856013101984 2013 eng d a0964-830500aApproach towards biocatalytic valorisation of barley beta-glucan for bioethanol production using 1,3-1,4 beta-glucanase and thermotolerant yeast0 aApproach towards biocatalytic valorisation of barley betaglucan  aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcAUG a81-860 v823 a

The exploitation of renewable resource containing polymers other than cellulose and hemicellulose are critically important for the feasibility of biofuel production. The potential of 1,3-1,4 glucan 4-gluconohydrolyase mediated saccharification of barley beta-glucan (BG) was investigated for ethanol production using thermotolerant Saccharomyces sp. A maximum hydrolysis of 71% was obtained in 24 h using in-house produced 1,3-1,4 beta-glucanase from an alkalothermophilic Thermomonospora sp. whereas the hydrolysis was 100% with Accellerase (TM) 1000. The synergistic effect of beta-glucosidase and 1,3-1,4 beta-glucanase was demonstrated by the exogenous addition of beta-glucosidase to Thermomonospora 1,3-1,4 beta-glucanase which resulted in complete hydrolysis of BG. The hydrolysates of BG obtained using Accellerase or a cocktail of Thermomonospora 1,3-1,4 beta-glucanase and beta-glucosidase when fermented with free cells of Saccharomyces at 40 degrees C produced an ethanol yield of 0.44 g g(-1) and 0.46 g g(-1) respectively and when fermented with immobilized cells produced a yield of 0.49 g g(-1). The Ca-alginate immobilized yeast cells were reused nine times at 40 degrees C with 100% fermentation efficiency. The economics of barley-to-fuel ethanol program will ameliorate if in addition to barley starch, beta-glucan is also utilized. (c) 2013 Elsevier Ltd. All rights reserved.

10a110a3-110a4 Glucan 4-gluconohydrolyase10aBarley beta-glucan10aEthanol10aHydrolysis10aSynergism10aThermotolerant yeast1 aDivate, Rupesh1 aMenon, Vishnu1 aRao, Mala uhttps://library.ncl.res.in/content/approach-towards-biocatalytic-valorisation-barley-beta-glucan-bioethanol-production-using-002035nas a2200229 4500008004100000022001400041245012600055210006900181260007500250300001200325490000700337520117600344653002701520653001601547653002001563653002701583653001401610653002301624100001801647700001401665856012601679 2013 eng d a1053-050900aMechanistic insights into the inhibition of endo-beta 1,4 xyloglucan hydrolase by a classical aspartic protease inhibitor0 aMechanistic insights into the inhibition of endobeta 14 xylogluc a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGER/PLENUM PUBLISHERScMAR a311-3210 v233 a

This is the first report of inactivation of xyloglucanase from Thermomonospora sp by pepstatin A, a specific inhibitor towards aspartic proteases. The steady state kinetics revealed a reversible, competitive, two-step inhibition mechanism with IC (50) and K (i) values of 3.5 +/- 0.5 mu M and 1.25 +/- 0.5 mu M respectively. The rate constants determined for the isomerization of EI to EI* and the dissociation of EI* were 14.5 +/- 1.5 x 10(-5) s(-1) and 2.85 +/- 1.2 x 10(-8) s(-1) respectively, whereas the overall inhibition constant K (i) (*) was 27 +/- 1 nM. The conformational changes induced upon inhibitor binding to xyloglucanase were monitored by fluorescence analysis and the rate constants derived were in agreement with the kinetic data. The abolished isoindole fluorescence of o-phthalaldehyde (OPTA)-labeled xyloglucanase and far UV analysis suggested that pepstatin binds to the active site of the enzyme. Our results revealed that the inactivation of xyloglucanase is due to the interference in the electronic microenvironment and disruption of the hydrogen-bonding network between the essential histidine and other residues involved in catalysis.

10a4 xyloglucan hydrolase10aEndo-beta 110aEnzyme kinetics10aInactivation mechanism10aPepstatin10aSlow-tight binding1 aMenon, Vishnu1 aRao, Mala uhttps://library.ncl.res.in/content/mechanistic-insights-inhibition-endo-beta-14-xyloglucan-hydrolase-classical-aspartic-002145nas a2200217 4500008004100000022001400041245014700055210006900202260007900271300001400350490000800364520126400372653002101636653002601657653002701683653003401710653002601744100001801770700001401788856012501802 2014 eng d a0273-228900aInteractions of a low molecular weight inhibitor from streptomyces sp. MBR04 with human cathepsin D: implications in mechanism of inactivation0 aInteractions of a low molecular weight inhibitor from streptomyc a999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USAbHUMANA PRESS INCcNOV a1705-17230 v1743 a

Cathepsin D, a lysosomal aspartic protease, is of potential interest as a target for drug design due to its implication in breast and ovarian cancer. The article reports a low molecular weight cathepsin D inhibitor from Streptomyces sp. MBR04. The M-r of the inhibitor was 1,078 Da as determined by MALDI-TOF, and the amino acid analysis showed the presence of Asp, Asp, Gly, Ala, Lys, Leu, Tyr, Trp residues. The steady-state kinetic interactions revealed reversible, competitive, slow-tight-binding nature of the inhibitor with an IC50 and K (i) values of 3.2 and 2.5 nM, respectively. The binding of the inhibitor with the enzyme and the subsequent conformational changes were monitored by exploiting the intrinsic fluorescence of the surface exposed Trp-54 residue. Based on the fluorescence and circular dichroism studies, we demonstrate that the inhibitor binds to the active site of cathepsin D and causes inactivation. All these kinetic, thermodynamic, and quenching studies suggest that the newly isolated peptidic inhibitor could be a potential scaffold to study and can be used to develop new potent therapeutic lead molecule for the development of drugs. The inhibitor will be significant as a potential lead molecule to target cathepsin D.

10aCancer treatment10aCathepsin D inhibitor10aInactivation mechanism10aSlow-tight-binding inhibition10aStreptomyces sp MBR041 aMenon, Vishnu1 aRao, Mala uhttps://library.ncl.res.in/content/interactions-low-molecular-weight-inhibitor-streptomyces-sp-mbr04-human-cathepsin-d-000523nas a2200157 4500008004100000022001400041245004400055210003900099260007500138300001200213490000700225100001400232700002300246700002100269856007500290 2016 eng d a0250-541X00aDr. V. Jagannathan (1921-2015) Obituary0 aDr V Jagannathan 19212015 Obituary a5 LAJPATRAI RD, ALLAHABAD 211002, INDIAbNATL ACAD SCIENCES INDIAcAPR a135-1360 v391 aRao, Mala1 aDeshpande, Vasanti1 aIngle, Prabhakar uhttps://library.ncl.res.in/content/dr-v-jagannathan-1921-2015-obituary