title: Purification and Properties of a Novel Xanthan Depolymerase from a Salt-Tolerant Bacterial Culture, HD1.
abstract: A novel xanthan depolymerase (endo-beta-1,4-glucanase) was isolated from a salt-tolerant bacteria culture (HD1) grown on xanthan. The depolymerase was purified 55-fold through chromatography on ion-exchange and molecular sieve columns, including high-performance liquid chromatography. The purified enzyme fraction was homogeneous as judged by polyacrylamide gel electrophoresis. The molecular weight of this enzyme is 60,000. Optimum pH and temperature for xanthan depolymerase activity were around 5 and 30 to 35 degrees C, respectively. The enzyme was not stable at a temperature higher than 45 degrees C. The activation energy calculated from an Arrhenius plot was 6.40 kcal (26.78 kJ). The enzyme molecule contains no sugar moiety. The amino acid composition of the enzyme protein was determined. Xanthan depolymerase cleaves the endo-beta-1,4-glucosidic linkage of the xanthan molecule, freeing reducing groups of some sugars and decreasing viscosity of the polymer solution. Only the backbones of beta-1,4-linked glucans with side chains or other substituents were cleaved. No monosaccharide was produced by the action of this enzyme. The oligosac-charide(s) in the low-molecular weight fraction consisted of 15 to 58 monosaccharide units. The enzymic reaction resulted in the decrease in weight-average molecular weight of xanthan from 6.5 x 10 to 8.0 x 10 in 0.5 h. This enzyme alone could not degrade xanthan to a single or multiple pentasaccharide unit(s). Results suggest that there may be regions inside the xanthan molecule that are susceptible to the attack of this enzyme. Xanthan depolymerase activity was not inhibited by many chemicals, including thiols, antioxidants, chlorinated hydrocarbons, metal-chelating agents, and inorganic compounds, except ferric chloride and arsenomolybdate. Many biocides were tested and found not to be inhibitory. Conditions used in enhanced oil recovery operations, i.e., the presence of formaldehyde, Na(2)S(2)O(4), 2,2-dibromo-3-nitrilopropionamide, and an anaerobic environment, did not inhibit xanthan depolymerase activity.
authors.affiliation: Corporate Research Sciences Laboratory, Exxon Research and Engineering Co., Annandale, New Jersey 08801.
authors.forename: C T
authors.initials: CT
authors.lastname: Hou
authors.forename: N
authors.initials: N
authors.lastname: Barnabe
authors.forename: K
authors.initials: K
authors.lastname: Greaney