Carrageenans are sulfated galactans found in the cell walls of numerous red seaweeds (Rhodophyta). They are classified according to the number and the position of sulfate ester groups and the occurrence of 3,6-anhydro-galactose. Although the carrageenan biosynthesis pathway is not fully understood, it is usually accepted that the last step consists of the formation of a 3,6-anhydro ring found in k-and i-carrageenans through the enzymatic conversion of D-galactose-6-sulfate or D-galactose-2,6-disulfate occurring in m-and n-carrageenan, respectively. We purified two enzymes, sulfurylase I (65 kD) and sulfurylase II (32 kD), that are able to catalyze the conversion of n-into i-carrageenan. We compared their sulfate release rates (i.e. arising from the formation of the anhydro ring) with the viscosity of the solution and demonstrated two distinct modes of action. In addition, we found that some mixtures of sulfurylase I and II lead to the formation of carrageenan solutions with unexpectedly low viscosities. We discuss the implication of these findings for the assembly of a densely aggregated matrix in red algal cell walls.Agars and carrageenans are the most abundant components of the cell walls in numerous red algae (Rhodophyta) and can represent up to 50% of algal dry weight. These sulfated galactans are densely packed in the cell wall in a three-dimensional solid network of pseudocrystalline fibers, which assemble during the deposition of cell wall macromolecules (Craigie, 1990).