中国食品添加剂2025，Vol.36Issue(7)：50-60,11.DOI:10.19804/j.issn1006-2513.2025.7.007
混菌发酵芡实制备抗氧化肽及其结构的鉴定
Preparation of antioxidant peptides from Euryale ferox via co-fermentation and their structural identification
摘要
Abstract
A co-culture fermentation system of Bacillus subtilis var.natto and Aspergillus oryzae on Euryale ferox substrates was established to produce highly active antioxidant peptides through multiscale optimization.A three-factor Box-Behnken response surface methodology(RSM)model was used to identified optimal fermentation parameters as follows:inoculum amounts of composite strains at 9.7％,volume ratio of strains(B.subtilis∶A.oryzae)at 2.2∶1,and fermentation time of(73±0.5)hours.Validation experiments demonstrated the peptide yield reached(73.72±0.13)％and protease activity was(198.79±0.94)U·mL-1.Structural identification revealed that antioxidant amino acids(Cys,Met,His)accounted for 66.24％of functional amino acids,hydrophobic amino acids made up 50.06％,and aromatic amino acids constituted 11.82％.The fraction E1(molecular weight≤1 kDa)exhibited significant antioxidant properties:DPPH·scavenging rate was(87.78±0.79)％(IC50=0.082 mg/mL),ABTS+·scavenging rate was(98.79±1.26)％(IC50=0.017 mg/mL),showing a 10.5％increase compared to glutathione(GSH)(P＜0.01);ferric reducing activity was measured by absorbance at 700 nm of(0.688±0.001)(IC50=1.97 mg/mL),with an efficiency coefficient(REC=0.58 mL·mg-2)increased 2.3-fold.MALDI-TOF/TOF mass spectrometry identified nine characteristic antioxidant peptides(411.37～1032.07 Da),with a core motif containing the Pro-Arg-Trp domain(accounting for 68％).Among them,a novel peptide segment IPILRW(m/z 743.382+)was found to bind Keapl protein with a binding free energy △ G=-32.6 kcal·mol-1.This study provides theoretical support for the industrial production of plant-derived antioxidants by elucidating the amino acid fingerprint characteristics and structure-activity relationships of Euryale ferox peptides.关键词
芡实/混菌发酵/抗氧化肽/结构鉴定Key words
Euryale ferox/mixed microbial fermentation/antioxidant peptide/structure identification分类
轻工纺织引用本文复制引用
毛绍云,柏倩,徐倩..混菌发酵芡实制备抗氧化肽及其结构的鉴定[J].中国食品添加剂,2025,36(7):50-60,11.基金项目
江苏省"青蓝工程"资助基金项目(JS1001) （JS1001）
徐州市推动科技创新项目(KC23051) （KC23051）
