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Enzymatic hydrolysis of penicillin mycelium protein
- Zhan'ao ZHANG Qingfen LIU
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Chin. J. Process Eng.. 2021, 21(4):
471-478.
DOI: 10.12034/j.issn.1009-606X.220122
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More than 2000000 tons of antibiotics fermentation residues (AFRs) were produced in China every year, which were hazardous solid waste, and how to safely dispose of them is attracting attention. AFRs are rich in organic substances, such as protein and carbohydrates, which accounts for about 50% of dry penicillin AFRs. So it is significant to develop new methods for recycling and utilization of mycelium protein in AFRs. In this work, penicillin mycelium was dissolved by alkali-thermal method, and penicillin mycelium protein solution was prepared, and then enzymatic hydrolysis performance of penicillin mycelium protein was explored in this solution using enzyme as catalyst, such as alcalase (A), bromelain (B), neutrase (C), and acid protease (D). The effects of types of enzymes, pH of the solution, the ratio of enzyme to protein, reaction temperature and time on protein hydrolysis process were systematically studied, and an optimization scheme was proposed. In a single enzyme-catalyzed hydrolysis process, alcalase showed the optimal performance, the hydrolysis degree of protein reached 31.43% at following conditions, the mass ratio of alcalase to protein as 6%, pH=11, the reaction time of 3 h and temperature of 50℃. In the complex enzyme-catalyzed hydrolysis process, the mixture of alcalase and bromelain as catalysts was demonstrated got the best efficiency. The optimum parameters were as follows: the mass ratio of complex enzyme to protein was 9% (A:B=2:1), pH=10, the reaction time of 3 h and temperature of 50℃. Under these optimal conditions, the protein hydrolysis degree reached 42.73%, which was 11.30 percentage points higher than the best single-enzyme method. The high performance liquid chromatography (HPLC) analysis found the enzymatic hydrolysate contained 16 amino acids, such as glutamic acid, aspartic acid, glycine, leucine and alanine, the ratio was 15.47wt%, 11.94wt%, 9.41wt%, 9.23wt%, 9.22wt%, respectively. This work provides a high efficient enzyme-catalyzed hydrolysis method of penicillin mycelium protein for preparing amino acids, and to carry out utilization of penicillin mycelium protein in AFRs.