In vitro synthesis of cellulose II from a cytoplasmic membrane fraction of Acetobacter xylinum

Journal Article · Thu Oct 01 04:00:00 EDT 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.20.6985· OSTI ID:7027201

Bureau, T E; Brown, Jr, R M

The cytoplasmic and outer membranes of Acetobacter xylinum were isolated by discontinuous sucrose density ultracentrifugation. Both lysozyme and trypsin were required for efficient crude membrane separation. Primary dehydrogenases and NADH oxidase were used as cytoplasmic membrane markers, and 2-keto-3-deoxyoctulosonic acid was used to identify the outer membranes. Cellulose synthetase activity was assayed as the conversion of radioactivity from UDP-(/sup 14/C)glucose into an alkali-insoluble ..beta..-1,4-D-(/sup 14/C)glucan. This activity was predominantly found in the cytoplasmic membrane. The cellulose nature of the product was demonstrated by (i) enzymatic hydrolysis followed by TLC, (ii) methylation analysis followed by TLC, and (iii) GC/MS. Further, the weight-average and number-average degree of polymerization of the in vitro product, determined by high-performance gel permeation chromatography, were 4820 and 5270, respectively. In addition, x-ray diffraction analysis indicated that the in vitro product is cellulose II, which is in contrast to the in vivo product--namely, cellulose I.

• Research Organization: Univ. of Texas, Austin (USA)
• OSTI ID: 7027201
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:20; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
550602 -- Medicine-- External Radiation in Diagnostics-- (1980-)
59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
ALDEHYDES
BIOCHEMISTRY
BIOSYNTHESIS
CARBOHYDRATES
CARBON 14 COMPOUNDS
CELL CONSTITUENTS
CELL MEMBRANES
CELLULOSE
CENTRIFUGATION
CHEMICAL REACTIONS
CHEMISTRY
CHROMATOGRAPHY
COHERENT SCATTERING
DECOMPOSITION
DIFFRACTION
ENZYMATIC HYDROLYSIS
ENZYMES
GAS CHROMATOGRAPHY
GLUCOSE
GLYCOSYL HYDROLASES
GLYCOSYL TRANSFERASES
HEXOSES
HEXOSYL TRANSFERASES
HYDROLASES
HYDROLYSIS
IN VITRO
LABELLED COMPOUNDS
LYSIS
LYSOZYME
MEMBRANES
MONOSACCHARIDES
O-GLYCOSYL HYDROLASES
ORGANIC COMPOUNDS
PEPTIDE HYDROLASES
POLYSACCHARIDES
SACCHARIDES
SCATTERING
SEPARATION PROCESSES
SERINE PROTEINASES
SOLVOLYSIS
SYNTHESIS
THIN-LAYER CHROMATOGRAPHY
TRANSFERASES
TRYPSIN
ULTRACENTRIFUGATION
X-RAY DIFFRACTION