Fermentation: Enzymes, Substrates, and SCFA

By: Chloe McClanahan, Biofiles Edition 6.2

The influence of probiotics on the metabolic activity of intestinal microbiota may be evaulated by detecting bacterial enzymatic activity in fecal samples, specifically β-glucosidase, β-glucuronidase, and urease activity. Experimental studies using probiotics have indicated a significant decrease in β-glucuronidase activity. A decrease in β-glucuronidase decreases the release of carcinogens from carcinogen-glucuronide conjugates within the colonic lumen.

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Enzymes

Name Specific Activity Unit Definition Form Cat. No.
β-Glucosidase from almonds ≥2 units/mg solid One unit will liberate 1.0 μmole of glucose from salicin per min at pH 5.0 at 37 °C. lyophilized powder
G0395-2.5KU
G0395-5KU
G0395-50KU
β-Glucosidase from almonds 10–30 units/mg solid lyophilized powder
G4511-100UN
G4511-250UN
G4511-1KU
β-Glucuronidase from Escherichia coli ≥20,000 units/mg protein One Sigma or modifi ed Fishman unit will liberate 1.0 μg of phenolphthalein from phenolphthalein glucuronide per hr at 37 °C at pH 6.8 (30 min assay). lyophilized powder
G8420-25KU
G8420-100KU
β-Glucuronidase from Escherichia coli 1,000,000–5,000,000 units/g protein lyophilized powder
G7396-25KU
G7396-125KU
G7396-250KU
G7396-500KU
G7396-1MU
G7396-2MU
β-Glucuronidase from Escherichia coli 5,000,000–20,000,000 units/g protein lyophilized powder
G7646-5KU
G7646-25KU
G7646-100KU
G7646-500KU
β-Glucuronidase from Escherichia coli 20,000,000–60,000,000 units/g protein aqueous glycerol solution
G8162-25KU
G8162-100KU
β-Glucuronidase from Escherichia coli 20,000,000–60,000,000 units/g protein lyophilized powder
G7896-25KU
G7896-100KU
G7896-200KU
β-Glucuronidase from Escherichia coli ≥10,000,000 units/g protein lyophilized powder
G8295-25KU
G8295-500KU
G8295-2MU
Urease from Canavalia ensiformis (Jack bean) 15,000–50,000 units/g solid One micromolar unit will liberate 1.0 μmole of NH 3 from urea per min at pH 7.0 at 25 °C. It is equivalent to 1.0 I.U. or 0.054 Sumner unit (1.0 mg ammonia nitrogen in 5 minutes at pH 7.0 at 20 °C) powder
U1500-20KU
U1500-100KU
U1500-500KU
U1500-1MU
Urease from Canavalia ensiformis (Jack bean) 50,000–100,000 units/g solid powder
U4002-20KU
U4002-100KU
U4002-500KU
U4002-1MU
Urease from Canavalia ensiformis (Jack bean) ≥600,000 units/g solid powder
U0251-6KU
U0251-12KU
U0251-60KU
Urease from Canavalia ensiformis (Jack bean) - glycerol solution
U1875-25ML
U1875-100ML

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Detection Substrates

Glucoside

Name Assay Cat. No.
3-Acetylumbelliferyl β-D -glucopyranoside ≥95% A3582-10MG
5-Bromo-4-chloro-3-indolyl β- D -glucopyranoside ≥97%
B4527-5MG
B4527-25MG
B4527-100MG
6-Bromo-2-naphthyl β- D -glucopyranoside ≥99%, TLC
B7877-1G
B7877-5G
 
6-Chloro-3-indolyl β- D -glucopyranoside ≥99.0%, HPLC 93546-25MG-F
Fluorescein di-(β- D -glucopyranoside) ≥90% F 4521
Indoxyl β- D -glucoside ≥97%
I3750-25MG
I3750-100MG
I3750-500MG
I3750-1G
I3750-2G
Indoxyl β- D -glucoside ≥97%
I6893-100MG
I6893-500MG
2-Methoxy-4-(2-nitrovinyl)phenyl β- D - glucopyranoside ≥95%
M2918-250MG
M2918-1G
4-Methylumbelliferyl α- D -glucopyranoside -
M9766-10MG
M9766-25MG
M9766-100MG
M9766-250MG
M9766-1G
4-Methylumbelliferyl β- D -glucopyranoside -
M3633-100MG
M3633-250MG
M3633-500MG
M3633-1G
M3633-5G
2-Nitrophenyl β- D -glucopyranoside -
N8016-250MG
N8016-1G
4-Nitrophenyl α-D -glucopyranoside ≥99%
N 1377-1G
N1377-5G
N1377-25G
4-Nitrophenyl β- D -glucopyranoside ≥98%, TLC
N7006-500MG
N7006-1G
N7006-5G
N7006-25G
Resorufin β- D -glucopyranoside ≥90% R 4758-50MG

Glucuronide

Name Assay Cat. No.
5-Bromo-4-chloro-3-indolyl β- D -glucuronide cyclohexylammonium salt ≥98% B3783-100MG
5-Bromo-4-chloro-3-indolyl β- D -glucuronide cyclohexylammonium salt ≥98%, TLC
B6650-10MG
B6650-25MG
B6650-100MG
5-Bromo-4-chloro-3-indolyl β- D -glucuronide cyclohexylammonium salt 10 mg substrate per tablet - B8049-5TAB
5-Bromo-4-chloro-3-indolyl β- D -glucuronide cyclohexylammonium salt ≥98%
B0522-10MG
B0522-100MG
5-Bromo-4-chloro-3-indolyl β- D -glucuronide sodium salt ≥98%
B5285-10MG
B5285-25MG
B5285-100MG
5-Bromo-4-chloro-3-indolyl β- D -glucuronide sodium salt 10 mg substrate per tablet - B8174-5TAB
5-Bromo-6-chloro-3-indolyl β- D -glucuronide cyclohexylammonium salt ≥98% B4532-10MG
6-Chloro-3-indolyl-β- D -glucuronide cyclohexylammonium salt ≥97.0%, HPLC
24907-10MG
24907-50MG
Indoxyl β- D -glucuronide cyclohexylammonium salt -
I7638-5MG
I7638-25MG
I7638-50MG
4-Methylumbelliferyl β- D -glucuronide hydrate ≥98%, TLC
M9130-25MG
M9130-100MG
M9130-250MG
M9130-1G
Naphthol AS-BI β- D -glucuronide -
N1875-25MG
N1875-100MG
N1875-250MG
N1875-500MG
4-Nitrophenyl β- D -glucuronide ≥98%, TLC
N1627-25MG
N1627-50MG
N1627-100MG
N1627-250MG
N1627-1G
N1627-2G
Phenolphthalein β- D -glucuronide -
P0501-25MG
P0501-100MG
P0501-500MG
Phenolphthalein β- D -glucuronide sodium salt -
P0376-25MG
P0376-100MG
P0376-250MG
P0376-1G
4-Trifl uoromethylumbelliferyl glucuronide potassium salt ≥98%, TLC T6410-2.5MG

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Short-Chain Fatty Acids (SCFA)

Probiotics and prebiotics enhance SCFA end product synthesis. An increase in SCFA production lowers the gastrointestinal pH thus improving pathogen resistance and stimulating epithelial cell proliferation. The predominant SCFA, butyrate, is the preferred substrate for colonocytes which likely contributes to a normal colonocyte population and a lower cancer risk. SCFA production can be measured with gas chromatography analysis on fecal samples or intestinal modeling systems. However, due to normal colon absorption of 95% of the SCFA metabolites, stable isotope technology is a superior method of analysis.

Non-Labeled Short-Chain Fatty Acids

Name Cat. No.
Acetic acid, ReagentPlus ®, ≥99%
A6283-100ML
A6283-500ML
A6283-2.5L
Butyric acid, ≥99%
B103500-5ML
B103500-100ML
B103500-500ML
Hexanoic acid, 99%
H12137-500ML
H12137-1L
Isobutyric acid, 99%
I1754-100ML
I1754-500ML
I1754-1L
Isovaleric acid, 99%
129542-100ML
129542-500ML
Propionic acid, reagent grade, ≥99%
P1386-500ML
P1386-1L
Valeric acid, ≥99%
240370-5ML
240370-100ML

Isotec® Short-Chain Fatty Acid Isotopes

Name Cat. No.
Sodium acetate-1-( 13 C), purum, 99 atom % 13 C 71198
Sodium acetate-1- 13 C, S & P tested, 99 atom % 13 C 668656
Sodium acetate-1- 13 C, 99 atom % 13 C 279293-1G
Sodium butyrate-1- 13 C, 99 atom % 13 C 292656
Sodium propionate-1- 13 C, 99 atom % 13 C 279455-1G

Materials

     
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