Objective To optimize the protein extraction process of Pectinidae. Methods A combination of composite enzymes and segmented enzymatic hydrolysis was employed, utilizing different protease enzymes with mode of enzyme digestion and cleavage sites. Significant factors were screened by single factor combined with Plackett-Burman and Box-Behnken model was used to optimize the one-stage protein extraction process, and the parameters of the two-stage enzymatic digestion process were optimized by orthogonal tests. Results The optimal extraction conditions for complex enzyme-assisted stepwise hydrolysis in protein extraction were as follows: The optimal extraction process parameters of the one-stage were as follows, solid-liquid ratio was 1:3.9 (m:V), enzyme digestion time was 80.0 min, compound enzyme addition was 1.1%, complex enzyme ratio (trypsin:alkaline protease) was 1:2 (m:m), enzyme digestion pH was 8.2, enzyme digestion temperature was 52.5 ℃; the optimal extraction parameters of the two-stage were as follows: Enzyme digestion pH was 7.0, enzyme digestion temperature was 50.0 ℃, neutral protease addition was 0.8%, and enzyme digestion time was 40 min. Under these conditions, the protein extraction rate was 76.3%. Conclusion Due to synergistic and complementary effects between the complex enzymes and segmented enzymatic hydrolysis, the protein extraction rate of Pectinidae is effectively improved, which provide some theoretical basis for the subsequent development and utilization of proteins from Pectinidae.
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |