Much progress was made in the straightforward and eco-friendly enzymatic synthesis of shorter cellulose chains (oligocellulose). Here, we report the determination of a molar mass distribution of the oligocellulose synthesized from cellobiose (CB) and α-glucose 1-phosphate by reverse phosphorolysis, using enzymes cellodextrin phosphorylase from Clostridium stercorarium or Clostridium thermocellum as catalyst. The oligocellulose molar mass distribution was analyzed using three different methods: (1)H NMR spectroscopy, matrix assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-ToF MS) and size exclusion chromatography (SEC). The molar mass distribution of the synthesized oligocellulose was only dependent on the concentration of cellobiose used in the reaction. Data obtained from MALDI-ToF MS and SEC were almost identical and showed that oligocellulose synthesized using 10 mM CB has an average degree of polymerization (DPn) of ∼7, while a DPn of ∼14 was achieved when 0.2 mM CB was used in the reaction. Because of solvent limitation in SEC analysis, MALDI-ToF MS was shown to be the technique of choice for accurate, easy and fast oligocellulose molar mass distribution determination.
DNA preparations from nuclear polyhedrosis virus (NPV) of Galleria mellonella L. (GmL) were fractionated in high ionic strength neutral sucrose gradient. This procedure allowed a separation of supercoiled infectious DNA molecules with contour length of 48--52 microns from infectious open ring DNA molecule, and noninfectious linear DNA molecules of the same size. In addition a heterogeneity of supercoiled DNA molecules was detected. Covalently closed DNA molecules did not contain protein or ribonucleotide ligands which could be digested by pronase or pancreatic RNase treatment. It is concluded from data on the infectivity of different molecular forms of DNA and reassociation kinetics studies, that the genome of GmL NPV is a unique ring nucleotide sequence with a molecular weight of about 90--100 X 10(6).
Foreign Science and Technology Center US Armay Materiel CQimeo1-Department o. thea Atuy INFECTIOUSNESS OF VIRAL DEOXY MJCLEIC ACIDS a. 06CRIPTIvIR Novae (I"* efft.wt ad kawh.9 de00)
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