Hydrolysis of wastewater colloidal organic matter by extracellular enzymes extracted from activated sludge flocs

2001 ◽  
Vol 43 (6) ◽  
pp. 33-40 ◽  
Author(s):  
A. Guellil ◽  
M. Boualam ◽  
H. Quiquampoix ◽  
P. Ginestet ◽  
J. M. Audic ◽  
...  

Enzymatic activities associated with the exopolymeric substances (EPSs) extracted from activated sludges were tested for their ability to hydrolyse the organic colloidal fraction of wastewater. Bacteria extracted with EPS and concentrated by wastewater microfiltration were inhibited with NaN3 or KCN. The protein hydrolysis mainly resulted from the enzymatic activity of EPS, whereas the glycolytic activity was mainly present in the organic colloidal fraction of the wastewater.

2016 ◽  
Vol 13 (9) ◽  
pp. 2815-2821 ◽  
Author(s):  
Federico Baltar ◽  
Catherine Legrand ◽  
Jarone Pinhassi

Abstract. Extracellular enzymatic activities (EEAs) are a crucial step in the degradation of organic matter. Dissolved (cell-free) extracellular enzymes in seawater can make up a significant contribution of the bulk EEA. However, the factors controlling the proportion of dissolved EEA in the marine environment remain unknown. Here we studied the seasonal changes in the proportion of dissolved relative to total EEA (of alkaline phosphatase (APase), β-glucosidase (BGase), and leucine aminopeptidase (LAPase)), in the Baltic Sea for 18 months. The proportion of dissolved EEA ranged between 37 and 100, 0 and 100, and 34 and 100 % for APase, BGase, and LAPase, respectively. A consistent seasonal pattern in the proportion of dissolved EEA was found among all the studied enzymes, with values up to 100 % during winter and  <  40 % during summer. A significant negative relation was found between the proportion of dissolved EEA and temperature, indicating that temperature might be a critical factor controlling the proportion of dissolved relative to total EEA in marine environments. Our results suggest a strong decoupling of hydrolysis rates from microbial dynamics in cold waters. This implies that under cold conditions, cell-free enzymes can contribute to substrate availability at large distances from the producing cell, increasing the dissociation between the hydrolysis of organic compounds and the actual microbes producing the enzymes. This might also suggest a potential effect of global warming on the hydrolysis of organic matter via a reduction of the contribution of cell-free enzymes to the bulk hydrolytic activity.


2016 ◽  
Author(s):  
F. Baltar ◽  
C. Legrand ◽  
J. Pinhassi

Abstract. Extracellular enzymatic activities (EEA) are a crucial step on the degradation of organic matter. Dissolved (cell-free) extracellular enzymes in seawater can make up a significant contribution of the bulk EEA. However, the factors controlling the proportion of dissolved EEA in the marine environment remain unknown. Here we studied the seasonal changes in the proportion of dissolved relative to total EEA (of alkaline phosphatase [APase], β-glucosidase, [BGase], and leucine aminopeptidase, [LAPase]), in the Baltic Sea for 18 months. The proportio n of dissolved EEA ranged between 37-100%, 0-100%, 34-100% for APase, BGase and LAPase, respectively. A consistent seasonal pattern in the proportion of dissolved EEA was found among all the studied enzymes, with values up to 100% during winter and <40% du ring summer. A significant negative relation was found between the 21proportion of dissolved EEA and temperature, indicating that temperature might be a critical factor controlling the proportion of dissolved relative to total EEA in marine environments. Our results suggest a strong decoupling of hydrolysis rates from mi crobial dynamics in cold waters. This implies that under cold conditions, cell-free enzymes can contribute to substrate availability at large distances from the producing cell, increasing the dissociation between the hydrolysis of organic compounds and the actual microbes producing the enzymes. This also indicates that global warming could come to affect the hydrolysis of organic matter by reducing the hydrolytic activity of cell-free enzymes.


2021 ◽  
Author(s):  
Greta Giljan ◽  
Sarah Brown ◽  
C. Chad Lloyd ◽  
Sherif Ghobrial ◽  
Rudolf Amann ◽  
...  

Heterotrophic bacteria use extracellular enzymes to hydrolyze high molecular weight (HMW) organic matter to low molecular weight (LMW) hydrolysis products that can be taken into the cell. These enzymes represent a considerable investment of carbon, nitrogen, and energy, yet the return on this investment is uncertain, since hydrolysis of a HMW substrate outside a cell yields LMW products that can be lost to diffusion and taken up by scavengers that do not produce extracellular enzymes1. However, an additional strategy of HMW organic matter utilization, selfish uptake2, is used for polysaccharide degradation, and has recently been found to be widespread among bacterial communities in surface ocean waters3. During selfish uptake, polysaccharides are bound at the cell surface, initially hydrolyzed, and transported into the periplasmic space without loss of hydrolysis products2, thereby retaining hydrolysate for the selfish bacteria and reducing availability of LMW substrates to scavenging bacteria. Here we show that selfish bacteria are common not only in the sunlit upper ocean, where polysaccharides are freshly produced by phytoplankton, but also deeper in the oceanic water column, including in bottom waters at depths of more than 5,500 meters. Thus, the return on investment, and therefore also the supply of suitable polysaccharides, must be sufficient to maintain these organisms.


1998 ◽  
Vol 38 (8-9) ◽  
pp. 179-188 ◽  
Author(s):  
K. F. Janning ◽  
X. Le Tallec ◽  
P. Harremoës

Hydrolysis and degradation of particulate organic matter has been isolated and investigated in laboratory scale and pilot scale biofilters. Wastewater was supplied to biofilm reactors in order to accumulate particulates from wastewater in the filter. When synthetic wastewater with no organic matter was supplied to the reactors, hydrolysis of the particulates was the only process occurring. Results from the laboratory scale experiments under aerobic conditions with pre-settled wastewater show that the initial removal rate is high: rV, O2 = 2.1 kg O2/(m3 d) though fast declining towards a much slower rate. A mass balance of carbon (TOC/TIC) shows that only 10% of the accumulated TOC was transformed to TIC during the 12 hour long experiment. The pilot scale hydrolysis experiment was performed in a new type of biofilm reactor - the B2A® biofilter that is characterised by a series of decreasing sized granular media (80-2.5 mm). When hydrolysis experiments were performed on the anoxic pilot biofilter with pre-screened wastewater particulates as carbon source, a rapid (rV, NO3=0.7 kg NO3-N/(m3 d)) and a slowler (rV, NO3 = 0.3 kg NO3-N/(m3 d)) removal rate were observed at an oxygen concentration of 3.5 mg O2/l. It was found that the pilot biofilter could retain significant amounts of particulate organic matter, reducing the porosity of the filter media of an average from 0.35 to 0.11. A mass balance of carbon shows that up to 40% of the total incoming TOC accumulates in the filter at high flow rates. Only up to 15% of the accumulated TOC was transformed to TIC during the 24 hour long experiment.


2016 ◽  
Vol 25 (2) ◽  
pp. 213 ◽  
Author(s):  
Kajar Köster ◽  
Frank Berninger ◽  
Jussi Heinonsalo ◽  
Aki Lindén ◽  
Egle Köster ◽  
...  

In boreal forest ecosystems fire, fungi and bacteria, and their interactions, have a pronounced effect on soil carbon dynamics. In this study we measured enzymatic activities, litter decomposition rates, carbon stocks and fungal and microbial biomasses in a boreal subarctic coniferous forest on a four age classes of non-stand replacing fire chronosequence (2, 42, 60 and 152 years after the fire). The results show that microbial activity recovered slowly after fire and the decomposition of new litter was affected by the disturbance. The percent mass loss of Scots pine litter increased with time from the last fire. Slow litter decomposition during the first post-fire years accelerates soil organic matter accumulation that is essential for the recovery of soil biological activities. Fire reduced the enzymatic activity across all the enzyme types measured. Carbon-degrading, chitin-degrading and phosphorus-dissolving enzymes showed different responses with the time elapsed since the fire disturbance. Microbial and enzymatic activity took decades before recovering to the levels observed in old forest stands. Our study demonstrates that slower post-fire litter decomposition has a pronounced impact on the recovery of soil organic matter following forest fires in northern boreal coniferous forests.


Microbiology ◽  
2004 ◽  
Vol 150 (7) ◽  
pp. 2257-2266 ◽  
Author(s):  
Helmuth Adelsberger ◽  
Christian Hertel ◽  
Erich Glawischnig ◽  
Vladimir V. Zverlov ◽  
Wolfgang H. Schwarz

Four extracellular enzymes of the thermophilic bacterium Clostridium stercorarium are involved in the depolymerization of de-esterified arabinoxylan: Xyn11A, Xyn10C, Bxl3B, and Arf51B. They were identified in a collection of eight clones producing enzymes hydrolysing xylan (xynA, xynB, xynC), β-xyloside (bxlA, bxlB, bglZ) and α-arabinofuranoside (arfA, arfB). The modular enzymes Xyn11A and Xyn10C represent the major xylanases in the culture supernatant of C. stercorarium. Both hydrolyse arabinoxylan in an endo-type mode, but differ in the pattern of the oligosaccharides produced. Of the glycosidases, Bxl3B degrades xylobiose and xylooligosaccharides to xylose, and Arf51B is able to release arabinose residues from de-esterified arabinoxylan and from the oligosaccharides generated. The other glycosidases either did not attack or only marginally attacked these oligosaccharides. Significantly more xylanase and xylosidase activity was produced during growth on xylose and xylan. This is believed to be the first time that, in a single thermophilic micro-organism, the complete set of enzymes (as well as the respective genes) to completely hydrolyse de-esterified arabinoxylan to its monomeric sugar constituents, xylose and arabinose, has been identified and the enzymes produced in vivo. The active enzyme system was reconstituted in vitro from recombinant enzymes.


2017 ◽  
Vol 14 (17) ◽  
pp. 3971-3977 ◽  
Author(s):  
Blair Thomson ◽  
Christopher David Hepburn ◽  
Miles Lamare ◽  
Federico Baltar

Abstract. Microbial extracellular enzymatic activity (EEA) is the rate-limiting step in the degradation of organic matter in the oceans. These extracellular enzymes exist in two forms: cell-bound, which are attached to the microbial cell wall, and cell-free, which are completely free of the cell. Contrary to previous understanding, cell-free extracellular enzymes make up a substantial proportion of the total marine EEA. Little is known about these abundant cell-free enzymes, including what factors control their activity once they are away from their sites (cells). Experiments were run to assess how cell-free enzymes (excluding microbes) respond to ultraviolet radiation (UVR) and temperature manipulations, previously suggested as potential control factors for these enzymes. The experiments were done with New Zealand coastal waters and the enzymes studied were alkaline phosphatase (APase), β-glucosidase, (BGase), and leucine aminopeptidase (LAPase). Environmentally relevant UVR (i.e. in situ UVR levels measured at our site) reduced cell-free enzyme activities by up to 87 % when compared to controls, likely a consequence of photodegradation. This effect of UVR on cell-free enzymes differed depending on the UVR fraction. Ambient levels of UV radiation were shown to reduce the activity of cell-free enzymes for the first time. Elevated temperatures (15 °C) increased the activity of cell-free enzymes by up to 53 % when compared to controls (10 °C), likely by enhancing the catalytic activity of the enzymes. Our results suggest the importance of both UVR and temperature as control mechanisms for cell-free enzymes. Given the projected warming ocean environment and the variable UVR light regime, it is possible that there could be major changes in the cell-free EEA and in the enzymes contribution to organic matter remineralization in the future.


2013 ◽  
Vol 10 (1) ◽  
pp. 567-582 ◽  
Author(s):  
S. Endres ◽  
J. Unger ◽  
N. Wannicke ◽  
M. Nausch ◽  
M. Voss ◽  
...  

Abstract. The filamentous and diazotrophic cyanobacterium Nodularia spumigena plays a major role in the productivity of the Baltic Sea as it forms extensive blooms regularly. Under phosphorus limiting conditions Nodularia spumigena have a high enzyme affinity for dissolved organic phosphorus (DOP) by production and release of alkaline phosphatase. Additionally, they are able to degrade proteinaceous compounds by expressing the extracellular enzyme leucine aminopeptidase. As atmospheric CO2 concentrations are increasing, we expect marine phytoplankton to experience changes in several environmental parameters, including pH, temperature, and nutrient availability. The aim of this study was to investigate the combined effect of CO2-induced changes in seawater carbonate chemistry and of phosphate deficiency on the exudation of organic matter, and its subsequent recycling by extracellular enzymes in a Nodularia spumigena culture. Batch cultures of Nodularia spumigena were grown for 15 days under aeration with low (180 μatm), medium (380 μatm), and high (780 μatm) CO2 concentrations. Obtained pCO2 levels in the treatments were on median 315, 353, and 548 μatm CO2, respectively. Extracellular enzyme activities as well as changes in organic and inorganic compound concentrations were monitored. CO2 treatment–related effects were identified for cyanobacterial growth, which in turn influenced the concentration of mucinous substances and the recycling of organic matter by extracellular enzymes. Biomass production was increased by 56.5% and 90.7% in the medium and high pCO2 treatment, respectively, compared to the low pCO2 treatment. In total, significantly more mucinous substances accumulated in the high pCO2 treatment, reaching 363 μg Xeq L−1 compared to 269 μg Xeq L−1 in the low pCO2 treatment. However, cell-specific rates did not change. After phosphate depletion, the acquisition of P from DOP by alkaline phosphatase was significantly enhanced. Alkaline phosphatase activities were increased by factor 1.64 and 2.25, respectively, in the medium and high compared to the low pCO2 treatment. We hypothesise from our results that Nodularia spumigena can grow faster under elevated pCO2 by enhancing the recycling of organic matter to acquire nutrients.


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