Table 4.
Substrate specificity profile of SPPS enzyme from Pleurotus sajor-caju strain CTM10057
| Substrate | Concentration | Absorbance (nm) a | Relative protease activity (%) b | ||
|---|---|---|---|---|---|
| Natural protein | Casein | 30 g/L | 660 | 100 ± 2.5 | |
| Gelatin | 30 g/L | 660 | 80 ± 2.0 | ||
| Keratin | 30 g/L | 660 | 37 ± 0.9 | ||
| Ovalbumin | 30 g/L | 660 | 20 ± 0.5 | ||
| BSA | 30 g/L | 660 | 10 ± 0.2 | ||
| Modified protein | Azo-casein | 25 g/L | 440 | 100 ± 2.5 | |
| Albumin azure | 25 g/L | 440 | 50 ± 1.8 | ||
| Keratin azure | 25 g/L | 595 | 30 ± 1.2 | ||
| Collagen type I | 20 g/L | 440 | 0 ± 0.0 | ||
| Collagen type II | 20 g/L | 440 | 0 ± 0.0 | ||
| Ester | ATEE | 10 mM | 253 | 100 ± 2.5 | |
| BTEE | 10 mM | 253 | 96 ± 2.4 | ||
| BAEE | 10 mM | 253 | 0 ± 0.0 | ||
| BCEE | 10 mM | 253 | 0 ± 0.0 | ||
| TAME | ↓ | 10 mM | 253 | 0 ± 0.0 | |
| Synthetic peptide (pNA) | Suc-F-pNA | 5 mM | 410 | 49 ± 1.5 | |
| Benz-Y-pNA | 5 mM | 410 | 0 ± 0.0 | ||
| M-pNA | 5 mM | 410 | 0 ± 0.0 | ||
| Ac-L-pNA | 5 mM | 410 | 0 ± 0.0 | ||
| P-pNA | 5 mM | 410 | 0 ± 0.0 | ||
| V-pNA | 5 mM | 410 | 0 ± 0.0 | ||
| Ac-A-pNA | 5 mM | 410 | 62 ± 1.6 | ||
| Benz-R-pNA | 5 mM | 410 | 0 ± 0.0 | ||
| Suc-YLV-pNA | 5 mM | 410 | 0 ± 0.0 | ||
| Suc-AAA-pNA | 5 mM | 410 | 50 ± 1.3 | ||
| Suc-AAV-pNA | 5 mM | 410 | 64 ± 1.7 | ||
| Suc-AAF-pNA | 5 mM | 410 | 62 ± 1.6 | ||
| Benz-FVR-pNA | 5 mM | 410 | 0 ± 0.0 | ||
| Suc-FAAF-pNA | 5 mM | 410 | 100 ± 2.5 | ||
| Suc-AAPF-pNA | 5 mM | 410 | 97 ± 2.5 | ||
| Suc-AAVA-pNA | 5 mM | 410 | 92 ± 2.4 | ||
| Suc-AAPM-pNA | 5 mM | 410 | 77 ± 1.9 | ||
| Suc-AAPL-pNA | 5 mM | 410 | 65 ± 1.7 | ||
| Suc-LLVY-pNA | 5 mM | 410 | 57 ± 1.6 | ||
| Ac-YVAD-pNA | 5 mM | 410 | 0 ± 0.0 | ||
a Values represent means of three independent replicates, and ± standard errors are reported
b The activity of each substrate was determined by measuring absorbance at specified wave lengths according to the relative method reported elsewhere [33, 34]
BTEE, N-benzol-l-tyrosine ethyl ester; ATEE, N-acetyl-l-tyrosine ethyl ester monohydrate; BAEE, N-benzol-l-arginine ethyl ester; BCEE, S-benzyl-l-cysteine ethyl ester hydrochloride; TAME, Nα-p-tosyl- l-arginine methyl ester hydrochloride; N-succinyl-F-p-nitroanilide; N-benzoyl-Y-p-nitroanilide; M-p-nitroanilide; N-acetyl-L-p-nitroanilide; P-p-nitroanilide trifluoroacetate salt; V-p-nitroanilide hydrochloride; N-acetyl-A-p-nitroanilide; N-benzoyl-R-p-nitroanilide; N-succinyl-YLV-p-nitroanilide; N-succinyl-AAA-p-nitroanilide; N-succinyl-AAF-p-nitroanilide; N-succinyl-AAV-p-nitroanilide; N-benzoyl-FVR-p-nitroanilide; N-succinyl-FAAF-p-nitroanilide; N-succinyl-AAPF-p-nitroanilide; N-succinyl-AAVA-p-nitroanilide; N-succinyl-AAPM-p-nitroanilide; N-succinyl-AAPL-p-nitroanilide; N-succinyl-LLVY-p-nitroanilide; and N-acetyl-YVAD-p-nitroanilide