Microbial processes, including extracellular enzyme (exoenzyme) production, are a major driver of decomposition and a current topic of interest in Arctic soils due to the effects of climate warming. While enzyme activity levels are often assessed, we lack information on the specific location of these exoenzymes within the soil matrix. Identifying the locations of different soil enzymes is needed to improve our understanding of microbial and overall ecosystem function. Using soil obtained from Utqiagvik, Alaska, our objectives in the study are (1) to measure the activity of enzymes in soil pore water, (2) to examine the distribution of activity among soil particle size fractions using filtration, and (3) to cross these particle size fraction analyses with disruption techniques (blending to shred and sonication to further separate clumped/ aggregated soil materials) to assess how tightly bound the enzymes are to the particles. The results of the soil pore water assays showed little to no enzyme activity (<0.05 nmol g soil(-1) h(-1)), suggesting that enzymes are not abundant in soil pore water. In the soil cores, we detected activity for most of the hydrolytic enzymes, and there were clear differences among the particle size and disruption treatments. Higher activities in unfiltered and 50-mu m filters relative to much finer 2-mu m filters suggested that the enzymes were preferentially associated with larger particles in the soil, likely the organic material that makes up the bulk of these Arctic soils. Furthermore, in the sonication + blending treatment with no filter, 5 of 6 hydrolytic enzymes showed higher activity compared to blending only (and much higher than sonication only), further indicating that enzyme-substrate complexes throughout the organic matter component of the soil matrix are the sites of hydrolytic enzyme activity. These results suggest that the enzymes are likely bound to either the producing microbes, which are bound to the substrates, or directly to the larger organic substrates they are decomposing. This close-proximity binding may potentially minimize the transport of decomposition products away from the microbes that produce them.

Climate change is expected to alter the mechanisms controlling soil organic matter (SOM) stabilization. Under climate change, soil warming and drying could affect the enzymatic mechanisms that control SOM turnover and dependence on substrate concentration. Here, we used a greenhouse climate manipulation in a mature boreal forest soil to test two specific hypotheses: (1) Rates of decomposition decline at lower substrate concentrations, and (2) reductions in soil moisture disproportionately constrain the degradation of low-concentration substrates. Using constructed soil cores, we measured decomposition rates of two polymeric substrates, starch and cellulose, as well as enzyme activities associated with degradation of these substrates. The greenhouse manipulation increased temperature by 0.8 A degrees C and reduced moisture in the constructed cores by up to 90 %. We rejected our first hypothesis, as the rate of starch decomposition did not decrease with declining starch concentration under control conditions, but we did find support for hypothesis two: Drying led to lower decomposition rates for low-concentration starch. We observed a threefold reduction in soil respiration rates in bulk soils in the greenhouses over a 4-month period, but the C losses from the constructed cores did not vary among our treatments. Activities of enzymes that degrade cellulose and starch were elevated in the greenhouse treatments, which may have compensated for moisture constraints on the degradation of the common substrate (i.e., cellulose) in our constructed cores. This study confirms that substrate decomposition can be concentration-dependent and suggests that climate change effects on soil moisture could reduce rates of decomposition in well-drained boreal forest soils lacking permafrost.