菌酶协同高效制备黑水虻蛋白肽及其生物活性研究

Optimization of Preparing Black Soldier Fly Larval Protein Peptides by Co-Fermented with Bacteria and Enzyme and Analysis of Biological Activity

为确定黑水虻虫浆蛋白肽的最佳制备条件,试验以新鲜黑水虻虫浆为原料,利用黑曲霉(Aspergillus niger)结合外源酶制剂胰蛋白酶作为菌酶发酵剂进行发酵。采用单因素和响应面法优化发酵工艺,通过单因素试验对接菌量、加水量、加酶量及发酵时间4个工艺参数进行研究,测定蛋白肽产量,在此基础上通过响应面法优化菌酶协同发酵工艺,建立回归预测模型。结果表明,黑水虻蛋白肽的最佳菌酶协同工艺条件为加水量32%、接菌量4.0%、加酶量0.1%、发酵时间35 h。在此条件下黑水虻蛋白肽产量可达295.57 mg/g,较未发酵组提升了92.0%,体外总抗氧化能力提升50.27%;黑水虻虫浆抗菌肽对金黄色葡萄球菌抑菌活性较未发酵组提升31.82%。说明菌酶协同制备的黑水虻蛋白肽具有更强的抗氧化活性,黑水虻抗菌肽具有更高的抑菌活性。

To determine the optimal conditions for the preparation of black soldier fly protein peptides,fresh black soldier fly worm slurry was used as the raw material in this experiment,and fermentation was carried out using Aspergillus niger combined with the exogenous enzyme preparation trypsin as the myco⁃enzymatic fermentation agent.The fermentation process was optimized using one-factor and response sur⁃face methodology,and four process parameters,namely,the amount of inoculum volume,the amount of water addition,the amount of enzyme addition and the fermentation time,were investigated and the pro⁃teolytic peptide content was determined by a one-factor test;on this basis,the microbiology enzyme cofermentation process was optimized by response surface methodology and regression prediction model was established.The results showed that the optimal bacterial-enzyme synergistic process conditions for black soldier fly peptide were 32%water addition,4.0%bacterial reception,0.1%enzyme addition,and 35 h fermentation time.Under this condition black soldier fly protein peptide content could reach 295.57 mg/g,which was 92.0%higher than that of the unfermented group,and the total antioxi⁃dant capacity in vitro was increased by 50.27%;the antimicrobial peptide of black soldier fly worm plasma had 31.82%higher antibacterial ac⁃tivity against Staphylococcus aureus than that of the unfermented group.It indicates that the black soldier fly protein peptide prepared by microbiol⁃ogy enzyme synergy has stronger antioxidant activ ity and the black soldier fly antimicrobial peptide has higher bacteriostatic activity.