Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1974 Nov;120(2):844–853. doi: 10.1128/jb.120.2.844-853.1974

Extracellular Enzyme From Myxobacter AL-1 That Exhibits Both β-1,4-Glucanase and Chitosanase Activities

Allan Hedges 1, R S Wolfe 1
PMCID: PMC245847  PMID: 4218236

Abstract

An enzyme that has both β-1,4-glucanase and chitosanase activities is characterized. Evidence for homogeneity was obtained from electrophoresis and sedimentation velocity studies; only one N-terminal amino acid, valine, was found. Results of denaturation studies showed that β-1,4-glucanase and chitosanase activities decreased at equal rates. With carboxymethylcellulose as the substrate, a Km of 1.68 g of carboxymethylcellulose per liter of solution and a Vmax of 2.20 × 10−9 mol/min were found. With chitosan (the β-1,4-polymer of glucosamine) as the substrate, a Km of 0.30 g of chitosan per liter of solution and a Vmax of 0.75 × 10−9 mol/min were found. A pH optimum of 5.0 was found for β-1,4-glucanase activity, and pH optima of 5.0 and 6.8 were found for chitosanase activity. β-1,4-Glucanase activity had a temperature optimum of 38 C, and chitosanase activity had a temperature optimum of 70 C. Chitosan stabilized both enzyme activities at 70 C. Cellotriose was the smallest polymer capable of hydrolysis. Glucosamine was released by action of the enzyme upon cell wall preparations of several fungi.

Full text

PDF
849

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arnott M. S., Ward D. N. Separation of dansyl amino acids in a single analysis. Anal Biochem. 1967 Oct;21(1):50–56. doi: 10.1016/0003-2697(67)90082-6. [DOI] [PubMed] [Google Scholar]
  3. ENSIGN J. C., WOLFE R. S. LYSIS OF BACTERIAL CELL WALLS BY AN ENZYME ISOLATED FROM A MYXOBACTER. J Bacteriol. 1965 Aug;90:395–402. doi: 10.1128/jb.90.2.395-402.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Goodwin T. W., Morton R. A. The spectrophotometric determination of tyrosine and tryptophan in proteins. Biochem J. 1946;40(5-6):628–632. doi: 10.1042/bj0400628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gros C., Labouesse B. Study of the dansylation reaction of amino acids, peptides and proteins. Eur J Biochem. 1969 Feb;7(4):463–470. doi: 10.1111/j.1432-1033.1969.tb19632.x. [DOI] [PubMed] [Google Scholar]
  6. Jackson R. L., Wolfe R. S. Composition, properties, and substrate specificities of Myxobacter AL-1 protease. J Biol Chem. 1968 Mar 10;243(5):879–888. [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. Monaghan R. L., Eveleigh D. E., Tewari R. P., Reese E. T. Chitosanase, a novel enzyme. Nat New Biol. 1973 Sep 19;245(142):78–80. doi: 10.1038/newbio245078a0. [DOI] [PubMed] [Google Scholar]
  9. Monreal J., Reese E. T. The chitinase of Serratia marcescens. Can J Microbiol. 1969 Jul;15(7):689–696. doi: 10.1139/m69-122. [DOI] [PubMed] [Google Scholar]
  10. PARK J. T., JOHNSON M. J. A submicrodetermination of glucose. J Biol Chem. 1949 Nov;181(1):149–151. [PubMed] [Google Scholar]
  11. TREVELYAN W. E., PROCTER D. P., HARRISON J. S. Detection of sugars on paper chromatograms. Nature. 1950 Sep 9;166(4219):444–445. doi: 10.1038/166444b0. [DOI] [PubMed] [Google Scholar]
  12. WHITAKER D. R. The mechanism of degradation of cellulose by Myrothecium cellulase. Can J Biochem Physiol. 1957 Sep;35(9):733–742. [PubMed] [Google Scholar]
  13. WOLFROM M. L., PATIN D. L., DELEDERKREMER R. M. THIN-LAYER CHROMATOGRAPHY ON MICROCRYSTALLINE CELLULOSE. J Chromatogr. 1965 Mar;17:488–494. doi: 10.1016/s0021-9673(00)99899-6. [DOI] [PubMed] [Google Scholar]
  14. Wingard M., Matsueda G., Wolfe R. S. Myxobacter AL-1 protease II: specific peptide bond cleavage on the amino side of lysine. J Bacteriol. 1972 Nov;112(2):940–949. doi: 10.1128/jb.112.2.940-949.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Yamane K., Suzuki H., Nisizawa K. Purification and properties of extracellular and cell-bound cellulase components of Pseudomonas fluorescens var. cellulosa. J Biochem. 1970 Jan;67(1):19–35. doi: 10.1093/oxfordjournals.jbchem.a129231. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES