Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Jun;61(6):2186–2192. doi: 10.1128/aem.61.6.2186-2192.1995

An Inhibitor-Based Method To Measure Initial Decomposition of Naturally Occurring Polysaccharides in Sediments

H Boschker, S A Bertilsson, E Dekkers, T E Cappenberg
PMCID: PMC1388462  PMID: 16535044

Abstract

A method that can be used to measure the initial decomposition rates of polysaccharides in sediment samples was developed. It uses toluene to specifically inhibit microbial uptake of carbohydrates without affecting extracellular hydrolysis of polysaccharides. Accumulating carbohydrates were determined by high-performance liquid chromatography. Field-sampled litter from the common reed (Phragmites australis), which contains cellulose and arabinoxylan as its main polysaccharides, was used as a model system. Toluene concentrations of between 1 and 10% resulted in the accumulation of similar amounts of monomeric carbohydrates, which was linear over time for most neutral sugars. Toluene (3%) did not have an effect on extracellular enzyme activities, and microbial sugar uptake was completely inhibited, as demonstrated with (sup14)C-labelled xylose and glucose. Experiments with enhancement cultures and fixed reed litter suggested that enzymatic hydrolysis of polysaccharides in reed litter was the main source of glucose, xylose, arabinose, and galactose accumulation. In contrast, the accumulation of high amounts of the alditols mannitol and glucitol was probably caused by lysis of the microbial population in toluene-treated reed litter. Glucose accumulated at rates of 1.3 and 0.10 (mu)mol (middot) g of dry matter content(sup-1) (middot) h(sup-1) under aerobic and anaerobic conditions, respectively, whereas xylose accumulation rates were only 10% of the glucose accumulation rates.

Full Text

The Full Text of this article is available as a PDF (303.7 KB).

Selected References

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

  1. Almin K. E., Eriksson K. E. Enzymic degradation of polymers. I. Viscometric method for the determination of enzymic activity. Biochim Biophys Acta. 1967 Jul 11;139(2):238–247. doi: 10.1016/0005-2744(67)90028-9. [DOI] [PubMed] [Google Scholar]
  2. Benner R., Maccubbin A. E., Hodson R. E. Preparation, characterization, and microbial degradation of specifically radiolabeled [C]lignocelluloses from marine and freshwater macrophytes. Appl Environ Microbiol. 1984 Feb;47(2):381–389. doi: 10.1128/aem.47.2.381-389.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boschker H. T., Cappenberg T. E. A Sensitive Method Using 4-Methylumbelliferyl-beta-Cellobiose as a Substrate To Measure (1,4)-beta-Glucanase Activity in Sediments. Appl Environ Microbiol. 1994 Oct;60(10):3592–3596. doi: 10.1128/aem.60.10.3592-3596.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bratbak G., Dundas I. Bacterial dry matter content and biomass estimations. Appl Environ Microbiol. 1984 Oct;48(4):755–757. doi: 10.1128/aem.48.4.755-757.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cunningham H. W., Wetzel Robert G. Kinetic Analysis of Protein Degradation by a Freshwater Wetland Sediment Community. Appl Environ Microbiol. 1989 Aug;55(8):1963–1967. doi: 10.1128/aem.55.8.1963-1967.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Felix H. Permeabilized cells. Anal Biochem. 1982 Mar 1;120(2):211–234. doi: 10.1016/0003-2697(82)90340-2. [DOI] [PubMed] [Google Scholar]
  7. Haginaka J., Nishimura Y., Wakai J., Yasuda H., Koizumi K., Nomura T. Determination of cyclodextrins and branched cyclodextrins by reversed-phase chromatography with pulsed amperometric detection and a membrane reactor. Anal Biochem. 1989 Jun;179(2):336–340. doi: 10.1016/0003-2697(89)90139-5. [DOI] [PubMed] [Google Scholar]
  8. King G. M. Characterization of beta-Glucosidase Activity in Intertidal Marine Sediments. Appl Environ Microbiol. 1986 Feb;51(2):373–380. doi: 10.1128/aem.51.2.373-380.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. King G. M., Klug M. J. Glucose metabolism in sediments of a eutrophic lake: tracer analysis of uptake and product formation. Appl Environ Microbiol. 1982 Dec;44(6):1308–1317. doi: 10.1128/aem.44.6.1308-1317.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Moran M. A., Hodson R. E. Bacterial secondary production on vascular plant detritus: relationships to detritus composition and degradation rate. Appl Environ Microbiol. 1989 Sep;55(9):2178–2189. doi: 10.1128/aem.55.9.2178-2189.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Sawyer T. E., King G. M. Glucose uptake and end product formation in an intertidal marine sediment. Appl Environ Microbiol. 1993 Jan;59(1):120–128. doi: 10.1128/aem.59.1.120-128.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Starink M., Krylova I. N., Bär-Gilissen M. J., Bak R. P., Cappenberg T. E. Rates of benthic protozoan grazing on free and attached sediment bacteria measured with fluorescently stained sediment. Appl Environ Microbiol. 1994 Jul;60(7):2259–2264. doi: 10.1128/aem.60.7.2259-2264.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Toerien D. F., Cavari B. Effect of temperature on heterotrophic glucose uptake, mineralization, and turnover rates in lake sediments. Appl Environ Microbiol. 1982 Jan;43(1):1–5. doi: 10.1128/aem.43.1.1-5.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wicks R. J., Moran M. A., Pittman L. J., Hodson R. E. Carbohydrate signatures of aquatic macrophytes and their dissolved degradation products as determined by a sensitive high-performance ion chromatography method. Appl Environ Microbiol. 1991 Nov;57(11):3135–3143. doi: 10.1128/aem.57.11.3135-3143.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Wong K. K., Tan L. U., Saddler J. N. Multiplicity of beta-1,4-xylanase in microorganisms: functions and applications. Microbiol Rev. 1988 Sep;52(3):305–317. doi: 10.1128/mr.52.3.305-317.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Yancey P. H., Clark M. E., Hand S. C., Bowlus R. D., Somero G. N. Living with water stress: evolution of osmolyte systems. Science. 1982 Sep 24;217(4566):1214–1222. doi: 10.1126/science.7112124. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES