The roles of oxygen and alginate-lyase in determining the molecular weight of alginate produced by Azotobacter vinelandii
An Azotobacter vinelandii mutant lacking alginate-lyase (SML2) and the wild type (ATCC 9046) were used to discriminate between the roles of the polymerase complex and alginate-lyase in the synthesis of alginate in cultures conducted under controlled dissolved oxygen tension (DOT). To avoid the presence of pre-synthesized alginates, all cultures were inoculated with washed cells. For cultures carried out at 3% DOT using the mutant, a well defined family of alginates of high mean molecular weight (MMW) were obtained (985 kDa). Under 1% and 5% DOT, the mutant produced unique families of alginates with lower MMW (150 and 388 kDa). A similar behavior was observed using the wild type: a production of well defined families of alginates of high MMW at 3% DOT (1,250 kDa) and lower MMW at 1% and 5% DOT (370 and 350 kDa). At the end of the ATCC 9046 fermentations, alginate was depolymerized by the action of lyases. Overall, the evidence indicated that polymerization of alginate is carried out by producing families of polysaccharide in a narrow MMW range, and that it is highly dependent on DOT. The role of alginate-lyase (present in the wild type) is restricted to a post-polymerization step.
This work was partially financed by DGAPA-UNAM (grant IN218201) and CONACyT (grant 39955). M.A. Trujillo-Roldán is indebted to DGEP-UNAM (Ph.D. scholarship) and to Foundation "Francisco José de Caldas" COLCIENCIAS-Colombia (M.Sc. scholarship). The authors thank Alejandra Rodríguez for assistance in conducting some of the cultures and to Dr. Carlos Peña, Dr. Guillermo Gosset and Dr. Gerardo Saucedo for helpful discussions and criticisms.
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