a) Soluble Dyed Substrates

1. The substrate is water soluble and is readily recognised and hydrolysed by an endo-acting hydrolase.

2. The reaction is terminated by the addition of a precipitant solution (e.g. ethanol) wherein high molecular weight, partially hydrolysed substrate fragments precipitate from solution.

3. The suspension is mixed thoroughly, centrifuged, and the colour in the solution is measured using a spectrophotometer.

4. The enzymatic activity is determined using a standard curve provided.

Our range of soluble dyed substrates

Product Name Product Code Enzyme Measured
Red Starch S-RSTAR α-Amylase
Red Debranched Arabinan (sugar beet) S-RDAR endo-1,5-α-Arabinanase
Azo-CM-Cellulose (liquid) S-ACMCL
Azo-CM-Cellulose (powder) S-ACMC endo-Cellulase / endo-1,4-β-Glucanase
Azo-Xyloglucan (tamarind) S-AZXG
Azo-Barley Glucan S-ABG100 Lichenase / endo-b-Glucanase
Azo-Fructan S-AZFR5 endo-Fructanase / endo-Inulinase
Azo-Fructan plus exo-Inulinase S-AZFRXOI
Red Pullulan S-RPUL Pullulanase / Limit-dextrinase
Azo-Carob Galactomannan S-ACGLM endo-1,4-β-Mannanase
Azo-Galactan (potato) S-AGALP endo-1,4-b-Galactanase
Azo-Casein (sulphanilamide dyed) S-AZCAS endo-Protease
AZ-Rhamnogalacturonan S-AZRH endo-Rhamnogalacturonan hydrolase / lyase
Azo-Wheat Arabinoxylan (liquid) S-AWAXL
Azo-Wheat Arabinoxylan (powder) S-AWAXP endo-1,4-b-Xylanase
Azo-Xylan (birchwood) (liquid) S-AXBL
Azo-Xylan (birchwood) (powder) S-AXBP

b) Insoluble Dyed Substrates

Megazyme’s insoluble dyed substrates are prepared by dyeing and crosslinking soluble polysaccharides.

These substrates are provided in both granular and fine particle size. The finely milled substrates are more useful in agar plate assays and semi quantitative tube and plate assays. Substrates in other colours are also being developed for gel/agar plate activity screenings.

Insoluble chromogenic substrates are recommended for semi-quantitative assays.

Key features of Megazyme’s insoluble dyed substrates
 Suitable for analysis of crude enzyme extracts  Quantitative assay
 Suitable for analysis of purified enzymes  Chemically defined substrate
 Suitable for gel/agar plate activity screening  Suitable for automation
 Suitable for endo-acting enzymes  Suitable for exo-acting enzymes

Assay procedure for insoluble dyed substrates
The hydrated insoluble substrate forms gelatinous particles which are readily recognised and hydrolysed by an endo-acting hydrolase. The partial hydrolysis of insoluble chromogenic substrates releases water soluble dyed substrate fragments.
The reaction is terminated by adding an alkaline solution to stop enzyme activity and the reaction slurry is filtered or centrifuged.
The colour released in solution is measured using a spectrophotometer and the colour intensity is directly related to enzyme activity.

Product Name Product Code Enzyme Measured
AZCL-Amylose (fine) I-AZAMYF a-Amylase
RedCL-Amylose (fine) I-RCLAMYF
AZCL-Arabinan (debranched) I-AZDAR endo-1,5-α-Arabinanase
AZCL-HE-Cellulose (fine) I-AZCELF
RedCL-HE-Cellulose (fine) I-RCLCELF
Azo-α-Cellulose I-ACELL endo-Cellulase
Azo-Avicel I-AAVIC
AZCL-Xyloglucan I-AZXYG
AZCL-Xyloglucan (fine) I-AZXYGF
RedCL-Xyloglucan (fine) I-RCLXYGF
AZCL-Chitosan I-AZCHAN endo-Chitosanase
AZCL-Chitosan (fine) I-AZCHANF
AZCL-Dextran I-AZDEX endo-1,6-α-Dextrinase
AZCL-Dextran (fine) I-AZDEXF
AZCL-Galactan (potato) I-AZGLP endo-1,4-b-Galactanase
AZCL-Galactan (potato) (fine) I-AZGLPF
AZCL-Curdlan (fine) I-AZCURF endo-1,3-b-Glucanase
RedCL-Curdlan (fine) I-RCLCURF
AZCL-Barley b-Glucan I-AZBGL
AZCL-Barley b-Glucan (fine) I-AZBGLF Lichenase / endo-b-Glucanase
RedCL-Barley b-Glucan (fine) I-RCLBGLF
AZCL-Galactomannan I-AZGMA
AZCL-Galactomannan (fine) I-AZGMAF endo-1,4-b-Mannanase
RedCL-Galactomannan (fine) I-RCLGMAF
AZCL-Casein I-AZCAS endo-Protease
AZCL-Pullulan (fine) I-AZPULF Pullulanase / Limit-Dextrinase
RedCL-Pullulan (fine) I-RCLPULF
AZCL-Rhamnogalacturonan 1 I-AZRHI Rhamnogalacturonan hydrolase / lyase
AZCL-Arabinoxylan (Wheat) I-AZWAX
AZCL-Arabinoxylan (Wheat) (fine) I-AZWAXF endo-1,4-b-D-Xylanase
AZCL-Xylan (Birchwood) I-AZXBW
RedCL-Arabinoxylan (Wheat) (fine) I-RCLWAXF

c) Tablet Substrates

Megazyme supplies a range of enzyme tablet tests for ultimate end-user convenience. The insoluble chromogenic substrates discussed above are formulated into tablets. By using tablet tests, it is possible to obtain quantitative determinations of endo-acting hydrolase activities.
From a procedural perspective, tablet tests operate in the same way as assays using insoluble chromogenic substrates. The advantage of enzyme tablet tests is that the need to accurately weigh substrate quantities (and the error associated with this parameter) is removed.

Key features of Megazyme’s tablet substrates
 Suitable for analysis of crude enzyme extracts  Chemically defined substrate
 Suitable for analysis of purified enzymes  Suitable for automation
 Quantitative assay  Suitable for gel/agar plate activity screening
 Suitable for endo-acting enzymes  Suitable for exo-acting enzymes

Our range of tablet substrates

Product Name Product Code Enzyme Measured
Amylazyme T-AMZ
Amylazyme BG T-AMZBG α-Amylase
Amylazyme Red T-AMZRD
Amylazyme HY T-AMZHY
Arabinazyme T-ARZ endo-1,5-α-Arabinanase
Cellazyme AF (40mg) T-CAF
Cellazyme C (60mg) T-CCZ endo-Cellulase / endo-1,4-b-Glucanase
Cellazyme T T-CTZ
Chitozyme T-CHZ endo-Chitosanase
α-Dextrazyme T-DEXT endo-1,6-α-Dextranase
b-Glucazyme T-BGZ Lichenase / endo-b-Glucanase
Galactazyme T-GLZ endo-1,4-b-Galactanase
1,3-b-Glucazyme T-PAZ endo-1,3-b-Glucanase
1,3-b-Glucazyme HS T-CUR
Limit-Dextrizyme T-LDZ Pullulanase / Limit-dextrinase
Mannazyme T-MNZ endo-1,4-b-Mannanase
Protazyme AK T-PRAK endo-Protease
Protazyme OL T-PROL
Rhamnozyme T-RHAM Rhamnogalacturonan hydrolase / lyase
Psyllazyme T-PSYL
Xylazyme AF (40mg) T-XAF endo-1,4-b-Xylanase
Xylazyme AX (60mg) T-XAX
Xylazyme (100mg) T-XYZ
Megazyme S-ABG100 麦芽粉中葡聚糖酶检测底物
Megazyme S-ACGLM 甘露聚糖酶检测底物
Megazyme S-AZFR5 内切型菊粉酶检测底物
Megazyme S-AZFRXOI 内切型菊粉酶特殊检测底物
Megazyme S-AGALP 半乳聚糖酶特殊检测底物
Megazyme S-AWAXL 木聚糖酶检测底物
Megazyme S-AWAXP 木聚糖酶检测底物(Powder)
Megazyme S-AXBL 木聚糖酶特殊检测底物
Megazyme S-AXBP 木聚糖酶特殊检测底物
Megazyme S-AZXG 纤维素酶可溶性发色检测底物
Megazyme S-AZRH 鼠李聚糖半乳糖醛酸内切酶特殊检测底物
Megazyme S-RDAR 阿拉伯聚糖检测底物
Megazyme S-RPUL 酶制备中的普鲁兰酶和麦粉中的极限糊精的检测底物
Megazyme S-RSTAR Alpha淀粉酶活性检测底物
Megazyme S-ACMC 葡聚糖酶检测底物
Megazyme S-ACMCL 葡聚糖酶检测底物(Liquid)
Megazyme I-AZAMY Alpha淀粉酶检测底物
Megazyme I-AZDAR 1,5-a-L-阿拉伯聚糖内切酶检测底物
Megazyme I-AZWAX 1,4-β-D-木聚糖内切酶检测底物
Megazyme I-AZBGL 麦芽Beta葡聚糖酶、地衣聚糖酶和纤维素酶检测底物
Megazyme I-AZCAS 蛋白酶检测底物
Megazyme I-AZCEL 纤维素内切酶检测底物
Megazyme I-AZCHAN 脱乙酰几丁质酶/脱乙酰壳多糖酶检测底物
Megazyme I-AZCOL 蛋白酶检测底物
Megazyme I-ACELL 纤维素内切酶检测底物
Megazyme I-AAVIC 纤维素内切酶检测底物
Megazyme I-AZPUL 极限糊精酶[普鲁兰酶]检测底物
Megazyme I-AZRHI 鼠李聚糖半乳糖醛酸水解酶和裂解酶检测底物
Megazyme I-AZXYG 纤维素内切酶检测底物
Megazyme I-AZBGLF AZCL-大麦β-葡聚糖(精细的)
Megazyme I-AZXBW 1,4-β-D-木聚糖内切酶检测底物.
Megazyme I-AZPAC 1,3-β-D-葡聚糖内切酶检测底物
Megazyme I-AZGMA 1,4-β-D-甘露聚糖内切酶检测底物
Megazyme I-AZGLP 1,4-β-D-半乳糖内切酶检测底物
Megazyme I-AZDEX 1,6-a-D-葡聚糖内切酶检测底物
Megazyme I-AZCUR 1,3-β-D-葡聚糖内切酶检测底物
Megazyme I-AZGLL AZCL-Galactan (Lupin) – 3g
Megazyme I-AZLIC AZCL-Lichenan 3g
Megazyme I-AZXOT AZCL-Xylan (Oat Spelt) 4g
Megazyme I-RCLAMYF RedCL-Amylose (fine) 5g
Megazyme I-RCLCELF RedCL-HE-Cellulose (fine) 3g
Megazyme I-RCLXYGF RedCL-Xyloglucan (fine) 4g
Megazyme I-AZGLPF AZCL-Galactan (Potato) (fine) 3g
Megazyme I-RCLGMAF RedCL-Galactomannan (fine) 3g
Megazyme I-AZAMYF AZCL-Amylose (fine) 5g
Megazyme I-AZPULF AZCL-Pullulan (fine) 3g
Megazyme I-AZDEXF AZCL-Dextran (fine) 3g
Megazyme I-AZCURF AZCL-Curdlan (fine) 3g
Megazyme I-AZXYGF AZCL-Xyloglucan (fine) 4g
Megazyme I-AZGMAF AZCL-Galactomannan (fine) 3g
Megazyme I-AZWAXF AZCL-Arabinoxylan (Wheat) (fine) 3g
Megazyme I-AZXBE AZCL-Xylan (Beechwood)
Megazyme S-AZCAS 蛋白酶底物(AZO-Casein)a