毛蚶蛋白对益生菌抗生素胁迫的保护作用

Protective Effect of Arca subcrenata Protein on Probiotics against Antibiotic Stress

长期摄入抗生素残留的水产品可引起肠道菌群稳态失调。该研究基于恩诺沙星对四种益生菌(两歧双歧杆菌BBi32、鼠李糖乳杆菌LGG、植物乳杆菌LP45和乳双歧杆菌Probio-M8)的抑菌作用,建立了抗生素胁迫的益生菌生长模型,评价了红三鱼、黄蚬和毛蚶蛋白粗提物及其不同分离纯化组分对益生菌抗生素胁迫的保护作用。在四种益生菌中,乳双歧杆菌Probio-M8对抗生素胁迫最为敏感,三种原料中毛蚶蛋白粗提物表现出对抗生素胁迫保护作用;毛蚶蛋白粗提物经过阴离子交换柱(SepharoseFastFlow)分离得到四个组分(Fr0、Fr1、Fr2和Fr3),其分子量分别为14.3~44.3 ku(Fr0)、20.1~29.0 ku(Fr1)、14.3~97.2 ku(Fr2)和<14.3 ku(Fr3),其中Fr0可更有效地提高Probio-M8菌液的OD600从0.80提升至1.56,Fr3可将其生长代时缩短至0.98 h。毛蚶蛋白可促进乳双歧杆菌在恩诺沙星胁迫下增殖的活性,表明毛蚶蛋白具有保护益生菌免受抗生素生长胁迫、促进益生菌增殖的功能,该研究为毛蚶蛋白对肠道稳态的调节功能提供理论依据,并提出了应对体内抗生素残留的应对方案。

Long-term intake of aquatic products with antibiotic residues can cause intestinal flora homeostasis imbalance.In this study,based on the inhibitory effects of enrofloxacin on four probiotics(Bifidobacterium bifidum BBi32,Lactobacillus rhamnosus LGG,Lactobacillus plantarum LP45 and Bifidobacterium lactis Probio-M8),a probiotic growth model for examining antibiotic stress was established to evaluate the protective effects of crude protein extracts of Nemipterus virgatus,Corbicula fluminea and Arca subcrenata as well as their purification fractions on the enrofloxacin-induced antibiotic stress towards probiotics.Among the four probiotics,Bifidobacterium lactis Probio-M8 showed the highest sensitivity to antibiotic stress.The Arca subcrenatacrude protein extract exhibited a protective effect against antibiotic stress among the three raw materials.Such a crude protein extract was then separated by an anion-exchange column to obtain four fractions(Fr0,Fr1,Fr2 and Fr3)with molecular weights of 14.3~44.3(Fr0),20.1~29.0(Fr1),14.3~97.2(Fr2)and<14.3(Fr3)ku,respectively.Among them,Fr0 showed greater effectiveness in increasing the OD600 of the Probio-M8 culture solution from 0.80 to 1.56,whilst Fr3 significantly shortened the growth generation time of Probio-M8 to 0.98 hour.The Arca subcrenata protein extract promoted the proliferation of Bifidobacterium lactis under enrofloxacin-induced stress;indicating that the protein extract is capable of protecting probiotic bacteria against antibiotic growth stress while promoting probiotic proliferation.This study provides a theoretical basis for the regulatory function of trichothecene protein on intestinal homeostasis and proposes a solution for dealing with antibiotic residues in vivo.