为筛选能够发酵棉籽糖蜜用于制备低聚糖合生元产品的益生菌菌株,研究以棉籽糖蜜为培养基,游离棉酚降解率为评判指标,筛选高效游离棉酚降解益生菌菌株。进一步对筛选的菌株进行安全性(溶血性及耐药性)、低聚糖保留能力、发酵性能(培养温度、生长曲线、产酸能力等)、益生性能(病原菌抑菌能力,耐酸和耐胆盐能力)进行研究。利用液相色谱-质谱联用技术对最佳菌株发酵棉籽糖蜜代谢产物进行分析。结果显示:从12株乳酸菌和15株酵母菌中共计筛选出3株对游离棉酚具有高效降解能力的菌株,分别为格氏乳杆菌(Lactobacillus gasseri)HX、植物乳植杆菌(Lactiplantibacillus plantarum)B6和瑞士乳杆菌(Lactobacillus helveticus)DLX,其游离棉酚降解率分别为81.10%、65.41%和79.56%。3株菌株都未出现溶血现象,并且对七类抗生素中的大部分种类都较为敏感;3株乳酸菌对低聚糖保留率在65%~95%,其中菌株HX对低聚糖保留能力最高;3株菌温度适应范围较广(20~42 ℃),但最适生长温度均为37 ℃;3株菌生长速度均较快,4~8 h达到对数生长期,并且活菌数即可达到109 CFU/mL;3株菌产酸能力较强,其中格氏乳杆菌HX产总酸能力最强达到了0.8%;3株菌对大肠杆菌、沙门氏菌和金黄色葡萄球菌都具有良好的抑菌活性;另外,3株菌均表现出较强的耐胆盐和耐酸性,其中HX耐酸性最高,在pH 2.0条件下4 h活菌数仍然能够保持在近80%,而pH≥3.0时活菌数超过100%。液相色谱-质谱联用分析结果显示菌株HX发酵后的棉籽糖蜜液中共检测到1 969种代谢物,其中含量较高的化合物为脂类及类脂质(35.41%)、有机杂环化合物(19.98%)、苯类(11.24%)、有机氮化合物(8.33%)、有机酸及其衍生物(8.19%)等。格氏乳杆菌HX具有高效的游离棉酚降解能力,安全,对低聚糖具有较高的保留能力,并且生长迅速、耐酸能力强、产代谢产物丰富,适合以棉籽糖蜜为原料发酵法制备低聚糖合生元产品。
This study aimed to screen probiotic strains capable of fermenting cottonseed molasses for oligosaccharide synbiotic production and systematically evaluate their safety, fermentation performance, and probiotic properties.Twelve lactic acid bacteria and fifteen yeast strains were screened for free gossypol degradation efficiency using cottonseed molasses medium.Selected strains underwent comprehensive evaluations, including safety assessment (hemolytic activity and antibiotic resistance), oligosaccharide retention capacity, fermentation characteristics (temperature adaptation, growth kinetics, acid production), and probiotic potential (pathogen inhibition, acid/bile salt tolerance).Metabolic profiling of the optimal strain’s fermentation products was performed via liquid chromatography-mass spectrometry (LC-MS).Three strains exhibiting high free gossypol degradation rates were identified:Lactobacillus gasseri HX (81.10%), Lactiplantibacillus plantarum B6 (65.41%), Lactobacillus helveticus DLX (79.56%).All strains demonstrated γ-hemolysis, susceptibility to seven antibiotic classes, and oligosaccharide retention rates of 65%-95% (peak retention by L.gasseri HX).They exhibited broad temperature adaptation (20-42 ℃) with optimal growth at 37 ℃, rapid logarithmic growth within 4-8 h (reaching 109 CFU/mL), and robust acid production (0.8% total acidity for L.gasseri HX).Pathogen inhibition against Escherichia coli, Salmonella, and Staphylococcus aureus was observed, alongside high bile salt tolerance and acid resistance.Notably, L.gasseri HX retained 80% viability after 4 h at pH 2.0 and exceeded 100% viability at pH≥3.0.LC-MS analysis detected 1,969 metabolites in L.gasseri HX-fermented molasses, predominantly lipids/lipid-like compounds (35.41%), organic heterocyclics (19.98%), benzenoids (11.24%), nitrogenous compounds (8.33%), and organic acids/derivatives (8.19%).L. gasseri HX demonstrates superior free gossypol degradation (81.10%), biosafety compliance, high oligosaccharide retention (95%), rapid proliferation, exceptional acid tolerance, and diverse metabolic output.These attributes validate its suitability as a starter culture for industrial production of cottonseed molasses-based synbiotics.The comprehensive metabolite profile provides critical insights for optimizing functional synbiotic formulations.
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