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Volume 271, Number 43, Issue of October 25, 1996 pp. 26843-26849
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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A Novel -(, , )-Glucanosyltransferase from the Cell Wall of Aspergillus fumigatus

(Received for publication, December 29, 1995, and in revised form, June 25, 1996)

Robbert P. Hartland , Thierry Fontaine , Jean-Paul Debeaupuis , Catherine Simenel ^¶ , Muriel Delepierre ^¶ and Jean-Paul Latgé

From  The Aspergillus Laboratory and the ^¶ Laboratory of Nuclear Magnetic Resonance, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France

Cell wall transferases utilizing -(, , )-glucan chains as substrates may play important roles in cell wall assembly and rearrangement, as -(, , )-glucan is a major structural component of the cell wall of many fungi. A novel -(, , )-glucanosyltransferase was purified to apparent homogeneity from an autolysate of the cell wall of Aspergillus fumigatus. The enzyme had a molecular mass of 49 kDa and contained approximately 5 kDa of N-linked carbohydrate. The enzyme catalyzed an initial endo-type splitting of a -(, , )-glucan molecule, followed by linkage of the newly generated reducing end to the nonreducing end of another -(, , )-glucan molecule. Laminarioligosaccharides of size G₁₀ and greater were donor substrates for the transferase. Laminarioligosaccharides of size G₅ and greater formed acceptors. The enzyme was able to reuse initial transferase products as donors and acceptors in extended incubations, resulting in the formation of increasingly larger transferase products until they became insoluble. The major initial products from an incubation of the transferase with borohydride-reduced G₁₁ (rG₁₁) were rG₆ and rG₁₆. ¹H NMR analysis of the rG₁₆ transferase product showed it was a laminarioligosaccharide, indicating that the enzyme forms a -(, , )-linkage during transfer. The enzyme may have a key function in vivo by allowing the integration of newly synthesized glucan into the wall and promoting cell wall expansion during cell growth.

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