抗龋齿口腔益生乳酸菌的筛选  

Screening of Oral Probiotic Lactic Acid Bacteria against Dental Caries

作　　者：宁亚维[1] 赵忠情 孙颖[1] 张东春 司海山 康亚朋 王志新[1] 王世杰 NING Yawei;ZHAO Zhongqing;SUN Ying;ZHANG Dongchun;SI Haishan;KANG Yapeng;WANG Zhixin;WANG Shijie(College of Food Science and Biology,Hebei University of Science and Technology,Shijiazhuang 050018,China)

机构地区：[1]河北科技大学食品与生物学院,河北石家庄050018

出　　处：《食品科学》2023年第10期195-204,共10页Food Science

基　　金：河北省十三五重点研发计划项目(20327125D);河北省自然科学基金项目(C2022208003)。

摘　　要：本研究旨在筛选出抑制口腔致病菌能力强、安全性高、口腔定植能力强的乳酸菌。首先,采用牛津杯琼脂扩散法从实验室乳酸菌菌种资源库中筛选抑制变异链球菌和戈氏链球菌的菌株,对抑菌性较高的28株乳酸菌菌株进行了溶血性评价,结果显示7株菌株呈现α-溶血,21株菌株无溶血性。通过中和处理、过氧化氢酶以及蛋白酶敏感性研究,探究无溶血性且对变异链球菌和戈氏链球菌抑菌圈直径≥10 mm的16株乳酸菌的抑菌代谢产物,筛选出7株抑菌能力强并有可能产生非有机酸类抑菌代谢产物的乳酸菌。安全性评价结果显示:7株乳酸菌产酸性以及对牙齿腐蚀性均较低,除菌株JZ14-1对溶菌酶最高耐受质量浓度为1.6 mg/mL,其他6株乳酸菌对溶菌酶最高耐受质量浓度为1.2 mg/mL,均可耐受溶菌酶。菌株HB13-2敏感的抗生素数量最多。口腔定植能力评价显示:菌株HB13-2具有最高的表面疏水性为44.00%,表面酸电荷为20.16%,表面碱电荷为49.44%,胞外多糖质量浓度为0.283 mg/mL;菌株HB13-2具有较低的自身生物膜形成能力和良好的自聚能力,易定植于口腔。菌株HB13-2对变异链球菌生物膜最高抑制率为59.72%。除此以外,具有较强的与变异链球菌共聚能力,在24 h共聚能力达到67.40%。综上,菌株HB13-2抑菌活性好、安全性高、口腔定植能力强,经16S rDNA测序鉴定为植物乳植杆菌,并命名为植物乳植杆菌(Lactiplantibacillus plantarum)HB13-2。The purpose of this study is to select lactic acid bacteria with a strong ability to inhibit oral pathogens,high safety and strong oral colonization capacity.Firstly,the strains that could inhibit Streptococcus mutans and Streptococcus gordonii were screened from lactic acid bacteria resource bank in the laboratory by Oxford cup diffusion method.The 28 selected strains were evaluated for hemolytic activity.The results showed that seven of the 28 strains showedα-hemolysis while the remaining 21 did not.The antibacterial metabolites of 16 strains without hemolysis and with a diameter of inhibition zone≥10 mm against S.mutans and S.gordonii were determined by neutralization,catalase and protease susceptibility assays.Then seven strains with strong bacteriostatic capacity and the potential to produce non-organic acid antimicrobial metabolites were selected.The safety evaluation results showed that all seven strains had low acid production ability and caused low corrosion to teeth.The highest tolerable concentration of lysozyme for strain JZ14-1 was 1.6 mg/mL,while that for the other six strains was 1.2 mg/mL,indicating that all seven strains could tolerate lysozyme.Strain HB13-2 was most susceptible to antibiotics.The oral colonization capacity evaluation of strain HB13-2 showed the following results:the highest hydrophobicity of 44.00%,surface acid charge of 20.16%,surface base charge of 49.44%,and content of exopolysaccharides of 0.283 mg/mL.This strain could easily colonize in oral cavity and had low biofilm formation capacity and good self-aggregation capacity,and the maximum inhibition percentage against S.mutans biofilm was 59.72%.Moreover,strain HB13-2 had strong copolymerization capacity with S.mutans,and the copolymerization capacity was 67.40%at 24 h.In conclusion,strain HB13-2 has good antibacterial activity,high safety and strong oral colonization capacity.It is identified as Lactiplantibacillus plantarum by 16S rDNA sequencing.

关 键 词：变异链球菌 乳酸菌 安全性评价 口腔定植能力 

分 类 号：TS201.3[轻工技术与工程—食品科学]