Isolation and Characterization of Microorganisms with Alternan Hydrolytic Activity

Purification of alternanase by affinity chromatography

“…NRRL B-21195 were obtained from the bacterial culture collection maintained at the National Center for Agricultural Utilization Research (Peoria, Ill.). The strain was grown at 30°C on a rotary shaker in a medium containing 1.7% Bacto tryptone, 0.3% Bacto Soytone (both from Difco, Detroit, Mich.), 0.5% NaCl, 0.25% K 2 HPO 4 , and 0.25% maltose; the pH of the medium was adjusted to 7.0 prior to autoclaving [4,16].…”

“…This soilderived bacteria was tentatively identified as a Bacillus sp. based on the formation of heat-tolerant spores and other diagnostic features [16]. Strain NRRL B-21195 constitutively produces alternanase, unlike other similar strains that require alternan in the medium for enzyme production.…”

Other microbial polysaccharides: pullulan, scleroglucan, elsinan, levan, alternant, dextran

“…mesenteroides NRRL B-1355. The native polymer can be produced in vivo by the intact bacterial culture or in vitro with cell-free supernatant containing alternansucrase (Leathers et al, 1996;Wyckoff et al, 1996;Monsan et al, 2001).…”

“…Alternan is recovered by precipitation with a suitable organic solvent as mentioned above. The product can be further purified and finally dried as discussed above (Leathers et al, 1996;Wyckoff et al, 1996;Monsan et al, 2001).…”

“…The crude product is further purified by dissolving it in water and re-precipitating it with ethanol or other suitable solvent. Finally, the product is dried by spray drying, oven drying or lyophilization (Leathers et al, 1996;Wyckoff et al, 1996;Monsan et al, 2001).…”