SOIL ENZYMES. Tento projekt je spolufinancován Evropským sociálním fondem a Státním rozpočtem ČR InoBio CZ.1.07/2.2.00/
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1 Soil Biology topic No. 11: SOIL ENZYMES
2 Enzymes Simple (only proteins) Composite protein + cofactor coenzyme (NAD +, NADP +, FAD +,FADP +, hem, α lipoate, Fe ions etc.) = holoenzyme) prosthetic group (coenzyme + apoenzyme Oligomeric enzymes (several enzymatic proteins)
3 Enzymes = biological catalysts High reaction rate Mild reaction conditions Reaction specifity Capacity for regulation Notes
4 Notes
5 Sites on enzyme molecules Notes Active site (catalytic and binding sites) Site for binding cofactor (composite enzymes) Stabilizing (activation) site Allosteric site (allosteric effectors)
6 Type of specifity for enzymes Notes Reaction specifity Substrate specifity
7 Classification Trivial names in (pepsin, trypsin) According to reaction ase (urease) International classification (substrate + type of reaction) urea aminohydrolase EC (cleavage of urea to NH 3 and CO 2 ) H 2 O 2 :H 2 O 2 oxidoreductase EC (H 2 O 2 + H 2 O 2 O 2 + 2H 2 O) (4 figure codes, class, subclass, group and serial number)
8 E + S ES E + P V Max v = V Max/2 v = Vmax [S] / (Km + [S]) K M [S] v = V Max [S] / (K M + [S])
9 Notes
10 Expression of enzymatic activity Notes mg nebo μmol product / unit of dry soil matter (g) / unit of time preriod (h) pkat product / per unit of DM
11 (vi) (V) Resting structure Living cell (ii) Death (iii) Lysis (vii) Dead intact cell Lysis (vi) (i) (I) Dead disintegrated cell Release Growth/Devision (iv) Secretion (vii) Extracellular enzymes (viii) Enzyme-substrate complex (vii) Clay minerals Living cells Humic acids Entraped enzymes (x) Adsorbed enzymes (ix) Humus-enzyme complex Clay-enzyme complex
12 Substrate Related: Adsorption Inhibition Stabilization Humification Site Related: Temperature Moisture Nutrients Enzyme Activity Microbial Production Decomposition Enzyme Production
13 Notes
14 Competetive (V Max = V Max ; K' M >K M ) Inhibitors Noncompetetive (V Max <V Max ;K' M =K M )
15 Notes
16 Soil enzymes Notes endoenzymes or intracellular exoenzymes, ectoenzymes or extracellular (biontic and abiontic)
17 Aplication of knowledge Notes Total biological activity Segment of microbial community High resolution
18 Enzymes
19 Dehydrogenases substrate H 2 + acceptor substrate + acceptor H 2 Catalase (hydrogen peroxidase oxidoreductase, EC ); H 2 O 2 H 2 O + O 2 Origin H 2 O 2 2O 2 + 2H + H 2 O 2 + O 2. L amino acid + H 2 O + E FAD(or E FMN) α keto acid + NH 3 + E FADH 2 E FADH 2 (or E FMNH 2 ) + O 2 E FAD (or E FMN) +H 2 O 2
20 Notes
21 Urease (urea amidohydrolase, EC )
22 Notes
23 L glutaminase ( L glutamine amidohydrolase, EC )
24 L asparaginase (L asparagine amidohydrolase EC )
25 Arylsulfase (arylsulfate sulfohydrolase, EC )
26 Notes
27 Proteases (EC 3.4.4)
28 Notes
29 Other enzymes Acid phosphomonoesterase (ortophosphoric monoester phosphohydrolase EC, ) β glucosidase (β D glucoside glucohydrolase, EC )
30 Tests of toxicity Microbiological Respiration an mineralization of substrates Ammonification and nitrification Activity of soil enzymes (urease, dehydrogenases, phosphatases) Test on soil fauna (e.g. springtails Folsomia candida, mites, earthworms Eisenia fetida, enchytraeid worms) Standardized tests e.g. different bacterial tests toxicity on microplates
31 Notes
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