02377nas a2200241   4500008004100000022001400041245011300055210006900168260007400237300001400311490000700325520145200332653004801784653001201832653002201844653002301866100001601889700001901905700002601924700002701950700002701977856013102004       2012                        eng d  a0301-485100aFunctional characterization and differential expression studies of squalene synthase from Withania somnifera0 aFunctional characterization and differential expression studies   aVAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDSbSPRINGERcSEP  a8803-88120 v393 a<p>Squalene synthase (SQS: EC 2.5.1.21) is a potential branch point regulatory enzyme and represents the first committed step to diverge the carbon flux from the main isoprenoid pathway towards sterol biosynthesis. In the present study, cloning and characterization of Withania somnifera squalene synthase (WsSQS) cDNA was investigated subsequently followed by its heterologous expression and preliminary enzyme activity. Two different types of WsSQS cDNA clones (WsSQS1and WsSQS2) were identified that contained an open reading frames of 1,236 and 1,242 bp encoding polypeptides of 412 and 414 amino acids respectively. Both WsSQS isoforms share 99 % similarity and identity with each other. WsSQS deduced amino acids sequences, when compared with SQS of other plant species, showed maximum similarity and identity with Capsicum annuum followed by Solanum tuberosum and Nicotiana tabacum. To obtain soluble recombinant enzymes, 24 hydrophobic amino acids were deleted from the carboxy terminus and expressed as 6X His-Tag fusion protein in Escherichia coli. Approximately 43 kDa recombinant protein was purified using Ni-NTA affinity chromatography and checked on SDS-PAGE. Preliminary activity of the purified enzymes was determined and the products were analyzed by gas chromatograph-mass spectrometer (GC-MS). Quantitative real-time PCR (qRT-PCR) analysis showed that WsSQS expresses more in young leaves than mature leaves, stem and root.</p>10aGas chromatograph-Mass Spectrometer (GC-MS)10aqRT-PCR10aSqualene synthase10aWithania somnifera1 aGupta, Neha1 aSharma, Poonam1 aKumar, R., J. Santosh1 aVishwakarma, Rishi, K.1 aKhan, Bashir, Mohammad  uhttp://library.ncl.res.in/content/functional-characterization-and-differential-expression-studies-squalene-synthase-withania-001772nas a2200229   4500008004100000022001400041245014100055210006900196260007500265300001200340490000700352520086000359653002401219653004301243653001201286653003401298653002201332100001901354700002401373700002601397856011901423       2013                        eng d  a0006-292800aCloning and sequence analysis of three variants of the gene encoding alkaline xylanase C from the alkaliphilic bacillus sp (NCL 87-6-10)0 aCloning and sequence analysis of three variants of the gene enco  a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGER/PLENUM PUBLISHERScOCT  a737-7490 v513 a<p>Alkaline xylanase C from the alkaliphilic Bacillus sp. (NCL 87-6-10) has a low molecular weight and alkaline pI and is cellulase-free, properties compatible with its use in the prebleaching of pulp. We report here the cloning and sequence analysis of three variants of the gene encoding xylanase C; xyl C1, xyl C2, and xyl C3. In phylogenetic analysis, the three xylanase C variants clustered into a single group along with other reported alkaline xylanases. Residues contributing to the alkaline pH were present in all three variants. DNA and protein sequence comparison of these variants with other reported alkaline xylanases revealed silent mutations, some of which are due to codon preference in the respective organisms. The recombinant Xyl C1 that was successfully expressed in E. coli BL21 (DE3) had properties similar to the native enzyme.</p>10aAlkaline xylanase C10aAlkaliphilic Bacillus sp (NCL 87-6-10)10aCloning10aE. coli BL21 (DE3) expression10aSequence analysis1 aSharma, Poonam1 aRele, Meenakshi, V.1 aKumar, Lalitha, Sunil  uhttp://library.ncl.res.in/content/cloning-and-sequence-analysis-three-variants-gene-encoding-alkaline-xylanase-c-0