01594nas a2200193   4500008004100000245013500041210006900176260000800245300001400253490000700267520084500274100001901119700002201138700002001160700003101180700003701211700002401248856012801272       2013                        eng d00aSelf-assembly to function: design, synthesis and broad spectrum antimicrobial properties of short hybrid E-vinylogous lipopeptides0 aSelfassembly to function design synthesis and broad spectrum ant  cNOV  a8468-84740 v563 aNonribosomal E-vinylogous gamma-amino acids are widely present in many peptide natural products and have been exploited as inhibitors for serine and cysteine proteases. Here, we are reporting the broad spectrum antimicrobial properties and self-assembled nanostructures of various hybrid lipopeptides composed of 1:1 alternating alpha-and E-vinylogous residues. Analysis of the results revealed that self-assembled nanostructures also play a significant role in the antimicrobial and hemolytic activities. In contrast to the alpha-peptide counterparts, vinylogous hybrid peptides displayed excellent antimicrobial properties against various bacterial and fungal strains. Peptides that adopted nanofiber structures displayed less hemolytic activity, while peptides that adopted nanoneedle structures displayed the highest hemolytic activity.1 aShankar, Shiva1 aBenke, Sushil, N.1 aNagendra, Narem1 aSrivastava, Prabhakar, Lal1 aThulasiram, Hirekodathakallu, V.1 aGopi, Hosahudya, N.  uhttp://library.ncl.res.in/content/self-assembly-function-design-synthesis-and-broad-spectrum-antimicrobial-properties-short02514nas a2200277   4500008004100000022001400041245011700055210006900172260000800241300001100249490000800260520162800268653001301896653002201909653001201931653001201943653002201955100001501977700001901992700002002011700001902031700002002050700001902070700002502089856012202114       2025                        eng d  a1046-592800aSimple freeze-thaw based method for efficient purification of recombinant human proinsulin from inclusion bodies0 aSimple freezethaw based method for efficient purification of rec  cMAR  a1066450 v2273 a<p>
	Insulin is a pivotal peptide hormone essential for regulating glucose homeostasis. It has been known for over 100 years, but its production and purification methods are still under improvement. Escherichia coli based bacterial expression system is primarily used for insulin production. The human insulin protein expressed in bacteria usually forms inclusion bodies, complicating the purification process. Traditionally, insulin purification is a timeconsuming process involving urea-based denaturation methods, and various refolding techniques, followed by extensive chromatographic methods. Here, we report an easy and efficient purification of human proinsulin involving freeze-thaw based solubilization method. The extracted proinsulin inclusion bodies are treated with different concentrations of urea, followed by a freeze-thaw based solubilization. The freezing was carried out at various temperatures, mainly -80 degrees C, -20 degrees C, and -196 degrees C to determine the optimum condition for solubilization. Highest solubilization of proinsulin from the inclusion body was achieved with 0.5M urea and -20 degrees C. Further Nickel NTA-based purification was performed, and the purified protein was characterized for disulfide mapping by high-resolution mass spectrometer (HRMS). We also performed glucose uptake assays to validate the functional properties of purified proinsulin. This freeze-thaw based mild solubilization approach is a fast and effective method for getting bioactive proinsulin, which will help further design better purification and processing strategies for insulin production.</p>
10aDiabetes10aDisulfide mapping10aglucose10ainsulin10aMass spectrometry1 aRajesh, S.1 aJathar, Swaraj1 aBanarjee, Reema1 aSharma, Monika1 aPalkar, Shivani1 aShankar, Shiva1 aKulkarni, Mahesh, J.  uhttp://library.ncl.res.in/content/simple-freeze-thaw-based-method-efficient-purification-recombinant-human-proinsulin