混菌发酵女贞子制备抗氧化肽及其结构的鉴定  

Preparation of antioxidative peptides from Ligustrum lucidum by mixed fermentation and identification of their structures

作　　者：柏倩[1] 毛绍云[1] 徐倩 BAI Qian;MAO Shaoyun;XU Qian(Jiangsu Cancer Hospital,Jiangsu institution of Cancer Research,The Affiliated Cancer Hospital of Nanjing Medical University,Nanjing 210009)

机构地区：[1]江苏省肿瘤医院,江苏省肿瘤防治研究所,南京医科大学附属肿瘤医院,南京210009

出　　处：《中国食品添加剂》2025年第2期34-42,共9页China Food Additives

摘　　要：以女贞子为原料,采用枯草芽孢杆菌和米曲霉为菌种混菌发酵制备抗氧化肽并对其结构进行鉴定。通过单因素和响应面试验对混菌发酵女贞子过程进行优化,最佳发酵条件为枯草芽孢杆菌和米曲霉体积比为2.5:1、复合菌种接种量为7.9%、发酵时间为73 h。该条件下蛋白酶活力达到(183.116±1.724)U/mL、多肽得率为(19.936±0.214)%,其中碱性氨基酸占比27.57%、抗氧化氨基酸占比62.28%。对发酵液进行超滤分析,结果发现分子质量≤1 kDa的C1组分对ABTS阳离子自由基、DPPH自由基清除率分别达到99.07%和85.79%,IC_(50)值分别达到(0.0985±0.018)和(2.17±0.007)mg/mL,铁离子还原力OD_(700)达到0.407。利用液相色谱-串联质谱法对C1组分进行多肽结构鉴定,获得以下LMVGLT、VPAPWTR、NRDDMRER、ANQLDQFSR、IGGIGTVPVGR和TVNSYNLPILRF 6条具有较强抗氧化活性的低聚肽。研究结果为女贞子高值化开发利用提供有力的实验支撑。The antioxidative peptides were prepared from Ligustrum lucidum through mixed fermentation of Bacillus subtilis and Aspergillus oryzae as microbial strains and the peptides structures were identified.The fermentation process of Ligustrum lucidum was optimized using single factor and response surface methodology.The optimal fermentation conditions were determined to be a volume ratio of Bacillus subtilis and Aspergillus oryzae of 2.5:1,an inoculation concentration of composite strain of 7.9%,and a fermentation time of 73 hours.Under these conditions,the protease activity was(183.116±1.724)U/mL,with a polypeptide yield of(19.936±0.214)%,Among the peptides,basic amino acids accounted for 27.57%,and antioxidative amino acids accounted for 62.28%.Ultrafiltration analysis of the fermentation broth showed that the fraction with a molecular weight≤1 kDa(C1)exhibited an ABTS cation radical scavenging rate of 99.07%and a DPPH radical scavenging rate of 85.79%,with IC_(50) values of(0.0985±0.018)and(2.17±0.007)mg/mL,respectively,and the reducing power of ferrous ions at OD_(700) reached 0.407.Liquid chromatography-tandem mass spectrometry( LC-MS/MS) was used for the structural identification of the C1 fraction,resulting in the identification of six highly antioxidative oligopeptides: LMVGLT、VPAPWTR、NRDDMRER、ANQLDQFSR、IGGIGTVPVGR and TVNSYNLPILRF. The results provide a strong experimental support for thehigh-value development and utilization of Ligustrum lucidum.

关 键 词：女贞子 混菌发酵 抗氧化肽 结构鉴定 

分 类 号：TS201[轻工技术与工程—食品科学]