Formation of D-glucose, cellobiose, and other saccharides during synthesis of cellulose by Acetobacter acetigenum in a lactate-buffered glycerol medium.

Particulars of Microorganism and Its Cultivation-The strain of A. czcetigenum used was NCIB 8132 from the National Collection of Industrial Bacteria, Department of Scientific and Industrial Research, Teddington, Middlesex, England. The defined medium, made with distilled water, contained (grams per liter): KHZPOI, 2.0; MgS04.7 HSO, 1.0; FeS04.7 HZO, 0.01; (NH&HP04, 2.5; calcium n-pantothenate, 0.002; riboflavin, 0.002; biotin, 0.0001; lactic acid, 10.3; and glycerol, 30.0. After dissolution of the lactic acid and salts the pH value was adjusted to 4.6 with potassium hydroxide followed by sterilization at 10 pounds per square inch for 15 minutes. Finally the vitamins in 2 ml. of aqueous ethanol (5C% by volume) were added. Ten liters of this medium were distributed among 40 Glaxo flasks and the organism grown in “shake culture” at 30” for 21 days. Paper Chromatography-The treated metabolism fluids (see below) were examined for their carbohydrate contents by paper chromatography. Throughout this work the following mixtures were employed : Solvent I, n-propanol-water-ethyl acetate, 7 :2 : 1, by volume (5), and Solvent II, ethyl acetate-pyridine-water, 2: 1:2, by volume (6). Reducing sugars were detected by aniline hydrogen phthalate reagent (7), and for detection of ketoses the napthoresorcinol-trichloroacetic acid spray of Partridge and Westall (8) was employed. Aniline-diphenylamine phosphate reagent was used to locate ketoses and aldoses on the same chromatogram (9). The chromatographic technique of Bayly and Bourne (10) was also employed. In this technique the oligosaccharides, after conversion to their benzylamine derivatives, are irrigated with the