Purification and some properties of the extracellular alpha-amylase-pullulanase produced by Clostridium thermohydrosulfuricum.
The novel alpha-amylase-pullulanase produced by Clostridium thermohydrosulfuricum E 101-69 was purified as two forms (I and II) from culture medium, by using gel filtration in 6 M-guanidine hydrochloride as the final step. Renatured alpha-amylase-pullulanase I and II had apparent Mr values of 370,000 +/- 85,000 and 330,000 +/- 85,000 respectively, as determined by native polyacrylamide-gradient-gel electrophoresis. Both forms appear to be dimers of two similar subunits, with Mr values of 190,000 +/- 30,000 for enzyme I and 180,000 +/- 30,000 for enzyme II according to SDS/polyacrylamide-gradient-gel electrophoresis. The two forms had similar amino acid compositions, the same N-terminal sequence (Glu-Ile-Asp-Thr-Ala-Pro-Ala-Ile) and the same pI of 4.25. Both forms contained sugars having mobilities identical with those of rhamnose, glucose, galactose and mannose. The amount of neutral hexoses relative to protein was 11-12% (w/w) for both forms.
Melasniemi
H
H
Research Laboratories, Finnish State Alcohol Company (Alko) Ltd., Helsinki.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D002850:Chromatography, Gel; D002855:Chromatography, Thin Layer; D003013:Clostridium / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D005110:Extracellular Space / Q000201:enzymology; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D007527:Isoenzymes / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D000516:alpha-Amylases / Q000302:isolation & purification*
D000596:Amino Acids; D002241:Carbohydrates; D007527:Isoenzymes; D006026:Glycoside Hydrolases; D000516:alpha-Amylases; C100181:pullulanase