Lactic acid bacteria(LAB)are important probiotics and also an important source of biological preservatives for preventing foodborne diseases.This study isolated Lactiplantibacillus plantarum 3-1 from traditional Chine...Lactic acid bacteria(LAB)are important probiotics and also an important source of biological preservatives for preventing foodborne diseases.This study isolated Lactiplantibacillus plantarum 3-1 from traditional Chinese pickles,which exhibited excellent broad-spectrum antibacterial activity,auto-aggregation ability(26.30%±1.44%),cell surface hydrophobicity(4.37%±0.77%),and safety among selected strains.Whole-genome sequencing identified a novel gene cluster encoding non-ribosomal peptide synthetase(NRPS),which serve as the genetic basis for the enhanced antibacterial activity of its fermentation supernatant.Transcriptome sequencing and a green fluorescent protein(gfp)-based promoter reporter system further confirmed the expression of this NRPS gene cluster.Optimization of the culture conditions(pH 6.0,0.4 mol/L sucrose,4%w/v NaCl)enhanced NRPS expression,as evidenced by a 3.65-fold and 2.10-fold increase in the antibacterial activity of the fermentation supernatant against Staphylococcus aureus and Escherichia coli,respectively.Then the crude NRPS-derived antimicrobial substances extract from the optimized culture effectively inhibited the growth of pathogenic bacteria in milk.In a word,this study highlights the potential probiotic properties of L.plantarum 3-1 and its NRPS-derived antibacterial compounds as effective bio-preservatives in the food industry.展开更多
调查我国7省12个不同地区的禾本科植物内生真菌,经分离、培养后鉴定得到2种Epichlo属和4种Neotyphodium属内生真菌。采用改良的QS(improved quick and safe,IQS)法进行禾本科植物内生真菌基因组DNA的提取,并通过PCR进行验证。以生长...调查我国7省12个不同地区的禾本科植物内生真菌,经分离、培养后鉴定得到2种Epichlo属和4种Neotyphodium属内生真菌。采用改良的QS(improved quick and safe,IQS)法进行禾本科植物内生真菌基因组DNA的提取,并通过PCR进行验证。以生长在平板上的菌丝为起始材料,仪器要求简单、操作简便,快速安全、省时省力,整个提取过程大约40 min。该方法和常规SDS法相比,时间缩短到2周,为原来的1/4。以IQS法提取的DNA为模板,检测禾本科植物内生真菌中的NRPS基因,并在NCBI上进行比对分析,发现与已有的NRPS基因高度相似。这种快速提取法所获得的基因组DNA质量高,可用于后续PCR、基因克隆及其他高通量测试,特别适合获取大量真菌菌株基因组DNA时使用。展开更多
基金supported by the National Natural Science Foundation of China(32272297)National Key Research and Development Program(2024YFF1107203)+4 种基金Xinjiang Uygur Autonomous Region Key Research and Development Program Project(2025B04032)Guangdong Province Foundation and Applied Basic Research Fund Project(2023A1515111145)Central Government Funds to Guide Local ScienceTechnology Development(2024ZY-JCYJ-02-45)Shaanxi Key R&D Plan Project-Key Industrial Innovation Chain Project(2023-ZDLNY-35,2024NC-ZDCYL-03-10)Shaanxi Science and Technology Innovation Team Project(2024RS-CXTD-75).
文摘Lactic acid bacteria(LAB)are important probiotics and also an important source of biological preservatives for preventing foodborne diseases.This study isolated Lactiplantibacillus plantarum 3-1 from traditional Chinese pickles,which exhibited excellent broad-spectrum antibacterial activity,auto-aggregation ability(26.30%±1.44%),cell surface hydrophobicity(4.37%±0.77%),and safety among selected strains.Whole-genome sequencing identified a novel gene cluster encoding non-ribosomal peptide synthetase(NRPS),which serve as the genetic basis for the enhanced antibacterial activity of its fermentation supernatant.Transcriptome sequencing and a green fluorescent protein(gfp)-based promoter reporter system further confirmed the expression of this NRPS gene cluster.Optimization of the culture conditions(pH 6.0,0.4 mol/L sucrose,4%w/v NaCl)enhanced NRPS expression,as evidenced by a 3.65-fold and 2.10-fold increase in the antibacterial activity of the fermentation supernatant against Staphylococcus aureus and Escherichia coli,respectively.Then the crude NRPS-derived antimicrobial substances extract from the optimized culture effectively inhibited the growth of pathogenic bacteria in milk.In a word,this study highlights the potential probiotic properties of L.plantarum 3-1 and its NRPS-derived antibacterial compounds as effective bio-preservatives in the food industry.
文摘调查我国7省12个不同地区的禾本科植物内生真菌,经分离、培养后鉴定得到2种Epichlo属和4种Neotyphodium属内生真菌。采用改良的QS(improved quick and safe,IQS)法进行禾本科植物内生真菌基因组DNA的提取,并通过PCR进行验证。以生长在平板上的菌丝为起始材料,仪器要求简单、操作简便,快速安全、省时省力,整个提取过程大约40 min。该方法和常规SDS法相比,时间缩短到2周,为原来的1/4。以IQS法提取的DNA为模板,检测禾本科植物内生真菌中的NRPS基因,并在NCBI上进行比对分析,发现与已有的NRPS基因高度相似。这种快速提取法所获得的基因组DNA质量高,可用于后续PCR、基因克隆及其他高通量测试,特别适合获取大量真菌菌株基因组DNA时使用。
