目的：

尿石素A是植物多酚的肠道代谢产物，具有抗炎、抗氧化、抗衰老及诱导线粒体自噬等重要生物活性。目前其合成路径主要依赖化学合成，但存在成本高、污染重等缺陷，而生物合成因环保且高效成为研究热点。然而，高效转化菌株的稀缺限制了其在功能性食品与药品中的应用。因此，本研究聚焦于植物多酚生物转化生成尿石素A的过程，进行了菌株代谢尿石素A的筛选、鉴定以及发酵工艺优化工作。希望通过融合微生物学、分析化学与生物工程技术，寻找到可以将植物多酚代谢生成尿石素A的菌株，为尿石素A的生物合成提供了新的菌种资源，为基于植物多酚的功能性食品及肠道健康干预产品的开发奠定了一定的理论基础。

方法：

本研究通过建立高效液相色谱法测定标准品尿石素A、尿石素B的保留时间，并利用内标法测量标准品峰面积来实现对尿石素A的定性定量。在此基础上，本研究通过单因素实验从石榴汁益生菌饮料中筛选到目标菌株，经平板划线纯化及 16S rDNA 测序，确定其菌属。

同时以 0.25–1.25 mg/ml 浓度的鞣花酸为底物，高效液相色谱仪为核心检测工具，结合酶标仪、pH 计等设备，初步构建菌种动力学体系。通过定期取样检测 pH、OD 值及尿石素 A 含量，系统探究菌株的代谢动力学特征，分析底物浓度与培养时间对菌株活性及尿石素 A 生成规律的影响，为深入解析菌株代谢机制提供数据支撑。

结果：

本研究首次从实验室自制的石榴汁益生菌饮料中分离并鉴定出具有高效转化能力的两种菌株，通过16S rDNA基因测序及系统发育树分析，确认目标菌株为植物乳杆菌和双歧杆菌的菌属，并分别命名为Urolithin A Lactobacillus、Urolithin A Bifidobacterium。进一步的代谢动力学研究揭示，在有氧条件下，Urolithin A Bifidobacterium对底物鞣花酸的亲和力（Kₘ=0.35 mM）明显高于Urolithin A Lactobacillus（Kₘ=0.62 mM），其最大反应速率Vₘₐₓ达到2.8 μM/h。本研究还创新性地发现，在无氧环境下，Urolithin A Bifidobacterium的代谢效率更佳：在0.5 mg/ml鞣花酸条件下，无氧组尿石素A的峰值（57.05 μM）显著高于有氧组（41.99 μM），且衰减速率对比Urolithin A Lactobacillus 更低。

结论：

本研究基于高效液相色谱技术，建立了尿石素A的定量分析方法，成功筛选并鉴定出高效转化尿石素 A 的Urolithin A Lactobacillus、Urolithin A Bifidobacterium，明确了其代谢特性及最优发酵条件。为尿石素 A 的生物合成提供了新方向。筛选的菌株符合食品安全标准，也为基于植物多酚的肠道健康干预产品开发奠定了基础，同时也为未来进一步探索代谢机制与工艺优化提供了理论依据。

Objective:

Urolithin A, a gut metabolite of plant polyphenols, possesses important biological activities such as anti-inflammatory, antioxidant, and anti-aging properties, as well as the ability to induce mitochondrial autophagy. Currently, its synthesis primarily relies on chemical methods, which are associated with high costs and environmental pollution. In contrast, biosynthesis has emerged as a prominent research focus due to its environmental sustainability and efficiency. However, the scarcity of highly efficient biotransforming strains has hindered its application in functional foods and pharmaceuticals. Therefore, this study focuses on the biotransformation of plant polyphenols into Urolithin A, involving the screening, identification, and optimization of fermentation processes for strains capable of metabolizing Urolithin A. By integrating microbiology, analytical chemistry, and bioengineering technologies, the research aims to identify strains that can metabolize plant polyphenols into Urolithin A, providing novel microbial resources for its biosynthesis and laying a theoretical foundation for the development of plant polyphenol-based functional foods and intestinal health intervention products..

Methods:

In this study, a High Performance Liquid Chromatography (HPLC) method was established to determine the retention times of Urolithin A and Urolithin B standards. The internal standard method was used to measure the peak areas of the standards, enabling qualitative and quantitative analysis of Urolithin A. On this basis, target strains were screened from pomegranate juice probiotic beverages through single-factor experiments. After streak plate purification and 16S rDNA sequencing, their bacterial genera were identified.

Using ellagic acid at concentrations of 0.25–1.25 mg/mL as the substrate, and with HPLC as the core detection tool, combined with equipment such as microplate readers and pH meters, a preliminary strain kinetic system was constructed. By regularly sampling to detect pH, OD values, and Urolithin A content, the metabolic kinetic characteristics of the strains were systematically investigated. The effects of substrate concentration and culture time on strain activity and the formation pattern of Urolithin A were analyzed, providing data support for in-depth analysis of the strain metabolic mechanisms

Results:

In this study, two highly efficient transformation strains were isolated and identified for the first time from laboratory-prepared pomegranate juice probiotic beverages. Through 16S rDNA gene sequencing and phylogenetic tree analysis, the target strains were confirmed to belong to the genera Lactobacillus plantarum and Bifidobacterium, and were named Urolithin A Lactobacillus and Urolithin A Bifidobacterium, respectively. Further metabolic kinetics studies revealed that under aerobic conditions, Urolithin A Bifidobacterium exhibited significantly higher affinity for the substrate ellagic acid (Kₘ = 0.35 mM) compared to Urolithin A Lactobacillus (Kₘ = 0.62 mM), with a maximum reaction rate (Vₘₐₓ) of 2.8 μM/h. This study also innovatively found that Urolithin A Bifidobacterium showed better metabolic efficiency under anaerobic conditions: at 0.5 mg/ml ellagic acid, the peak concentration of Urolithin A in the anaerobic group (57.05 μM) was significantly higher than that in the aerobic group (41.99 μM), and the decay rate was lower compared to Urolithin A Lactobacillus.

Conclusions：

Based on High Performance Liquid Chromatography (HPLC) technology, this study established a quantitative analytical method for Urolithin A, successfully screened and identified Urolithin A Lactobacillus and Urolithin A Bifidobacterium with high efficiency in biotransforming Urolithin A, and clarified their metabolic characteristics and optimal fermentation conditions. The research provides a new direction for the biosynthesis of Urolithin A. The screened strains comply with food safety standards, laying a foundation for the development of plant polyphenol-based intestinal health intervention products. Meanwhile, it offers a theoretical basis for future exploration of metabolic mechanisms and process optimization.

[1]Pandey,K.B.,Rizvi,S.I.(2009).Plant polyphenols as dietary antioxidants in human health and disease[J].Oxidative Medicine and Cellular Longevity,2(5),270-278.

[2]Shen N,Wang T,Gan Q,et al.(2022).Plant flavonoids:Classification,distribution,biosynthesis,and antioxidant activity[J].Food Chem.383:132531.

[3]Liu,Y.,Wang,P.,Chen,F.,et al.(2015).Role of plant polyphenols in acrylamide formation and elimination[J].Food chemistry,186,46-53.

[4]Iqbal,I.,Wilairatana,P.,Saqib,F.,et al.(2023).Plant Polyphenols and Their Potential Benefits on Cardiovascular Health:A Review[J].Molecules (Basel,Switzerland),28(17),64-03.

[5]Bouarab Chibane,L.,Degraeve,P.,Ferhout,et al.(2019).Plant antimicrobial polyphenols as potential natural food preservatives[J].Journal of the science of food and agriculture,99(4).

[6]Tsao R.(2010).Chemistry and biochemistry of dietary polyphenols[J].Nutrients,2(12),1231-1246.

[7]Singh,S.,Kaur,I.,&Kariyat,R.(2021).The Multifunctional Roles of Polyphenols in Plant-Herbivore Interactions[J].International journal of molecular sciences,22(3),1442.

[8]Garcia-Alonso,A.,Sánchez-Paniagua López,M.,Manzanares-Palenzuela,C.L.,et al.(2023).Edible plant by-products as source of polyphenols:prebiotic effect and analytical methods[J].Critical reviews in food science and nutrition,63(31),10814–10835.

[9]D'Amico,D.,Andreux,P.A.,Valdés,P.,et al.(2021).Impact of the Natural Compound Urolithin A on Health,Disease,and Aging[J].Trends in molecular medicine,27(7),687-699.

[10]D'Amico,D.,Andreux,P.A.,Valdés,P.,et al.(2021).Impact of the Natural Compound Urolithin A on Health,Disease,and Aging[J].Trends in molecular medicine,27(7),687-699.

[11]Cerdá,B.(2003).大鼠重复口服高剂量石榴ET安石榴单宁37天无毒[J].农业与食品化学杂志,51(11),3493-3501.

[12]Cerdá,B.,Llorach,R.,Cerón,J.J.,et al.(2003).Evaluation of the bioavailability and metabolism in the rat of punicalagin,an antioxidant polyphenol from pomegranate juice[J].European journal of nutrition,42(1),18-28.

[13]García-Villalba,R.,Giménez-Bastida,J.A.,Cortés-Martín,A.,et al.(2022).Urolithins:a Comprehensive Update on their Metabolism,Bioactivity,and Associated Gut Microbiota[J].Molecular nutrition &food research,66(21),e2101019.

[14]Garcia-Munoz,C.,&Vaillant,F.(2014).Metabolic fate of ellagitannins:implications for health,and research perspectives for innovative functional foods[J].Critical reviews in food science and nutrition,54(12),1584-1598.

[15]Zhang,M.,Cui,S.,Mao,B.,et al.(2023).Ellagic acid and intestinal microflora metabolite urolithin A:A review on its sources,metabolic distribution,health benefits,and biotransformation[J].Critical reviews in food science and nutrition,63(24),6900-6922.

[16]García-Villalba,R.,Giménez-Bastida,J.A.,Cortés-Martín,A.,et al.(2022).Urolithins:a Comprehensive Update on their Metabolism,Bioactivity,and Associated Gut Microbiota[J].Molecular nutrition &food research,66(21),e2101019.

[17]Tomás-Barberán,F.A.,Selma,M.V.,&Espín,J.C.(2016).Interactions of gut microbiota with dietary polyphenols and consequences to human health[J].Current Opinion in Clinical Nutrition &Metabolic Care,19(6),471-476.

[18]Haddad,E.H.,Gaban-Chong,N.,Oda,K.,&Sabaté,J.(2014).Effect of a walnut meal on postprandial oxidative stress and antioxidants in healthy individuals[J].Nutrition journal,13,4.

[19]Hidekazu,Ishimoto,Mari,et al.(2011).In vivo anti-inflammatory and antioxidant properties of ellagitannin metabolite urolithin a[J].Bioorganic &Medicinal Chemistry Letters,21(19),5901-5904.

[20]Guillermo Cásedas,Les,F.,Choya-Foces,C.,et al.(2020).The metabolite urolithin-a ameliorates oxidative stress in neuro-2a cells,becoming a potential neuroprotective agent[J].Antioxidants,9(2),177.

[21]Haddad,E.H.,Gaban-Chong,N.,Oda,K.,et al.(2014).Effect of a walnut meal on postprandial oxidative stress and antioxidants in healthy individuals[J].Nutrition Journal,13(1),4.

[22]Verzelloni,E.,Pellacani,C.,Tagliazucchi,D.,et al.(2011).Antiglycative and neuroprotective activity of colon-derived polyphenol catabolites[J].Molecular Nutrition &Food Research,55(S1),S35-S43.

[23]Li,X.,Zhang,Z.,Xu,G.,et al.(2024).Synthesis of urolithin derivatives and their anti-inflammatory activity[J].Biochemical and biophysical research communications,704,149711.

[24]D'Amico,D.,Olmer,M.,Fouassier,et al.(2022).Urolithin a improves mitochondrial health,reduces cartilage degeneration,and alleviates pain in osteoarthritis[J].Aging Cell,21(8).

[25]Tuohetaerbaike,B.,Zhang,Y.,Tian,Y.,et al.(2019).Pancreas protective effects of urolithin a on type 2 diabetic mice induced by high fat and streptozotocin via regulating autophagy and akt/mtor signaling pathway[J].Journal of ethnopharmacology,250.

[26]González-Sarrías,A.,Larrosa,M.,Tomás-Barberán,F.A.,et al.(2010).NF-kappaB-dependent anti-inflammatory activity of urolithins,gut microbiota ellagic acid-derived metabolites,in human colonic fibroblasts[J].The British journal of nutrition,104(4),503-512.

[27]Antonio González-Sarrías,Juan-Carlos Espín,Francisco A.Tomás-Barberán,et al.(2010).Gene expression,cell cycle arrest and mapk signalling regulation in caco-2 cells exposed to ellagic acid and its metabolites,urolithins[J].Molecular Nutrition &Food Research,53(6),686-698.

[28]王洪哲,谢海棠,徐乌兰.(2024).尿石素a通过活化mir-136介导的sirt1信号通路减轻呼吸道合胞病毒诱导的新生小鼠肺部感染[J].南方医科大学学报(7).

[29]Stanisławska,I.J.,Granica,S.,Piwowarski,J.P.,et al.(2019).The Activity of Urolithin A and M4 Valerolactone,Colonic Microbiota Metabolites of Polyphenols,in a Prostate Cancer In Vitro Model[J].Planta medica,85(2),118-125.

[30]田弥.(2023).尿石素a抗癌作用及其机制研究进展[J].药物流行病学杂志.

[31]D'Amico,D.,Andreux,P.A.,Valdés,P.,et al.(2021).Impact of the Natural Compound Urolithin A on Health,Disease,and Aging[J].Trends in molecular medicine,27(7),687-699.

[32]Al-Fatlawi,I.N.A.A.,Pouresmaeil,V.,Davoodi-Dehaghani,F.,et al.(2024).Effects of solid lipid nanocarrier containing methyl urolithin A by coating folate-bound chitosan and evaluation of its anti-cancer activity[J].BMC biotechnology,24(1),18.

[33]Zhang,M.,Cui,S.,Mao,B.,et al.(2023).Urolithin A Produced by Novel Microbial Fermentation Possesses Anti-aging Effects by Improving Mitophagy and Reducing Reactive Oxygen Species in Caenorhabditis elegans[J].Journal of agricultural and food chemistry,71(16),6348-6357.

[34]Kim,K.B.,Lee,S.,Kim,J.H.(2020).Neuroprotective effects of urolithin A on H2O2-induced oxidative stress-mediated apoptosis in SK-N-MC cells[J].Nutrition research and practice,14(1),3-11.

[35]Jiménez-Loygorri,J.I.,Villarejo-Zori,B.,Viedma-Poyatos,Á.,et al.(2024).Mitophagy curtails cytosolic mtDNA-dependent activation of cGAS/STING inflammation during aging[J].Nature communications,15(1),830.

[36]Martins,R.N.(2021).Therapeutic potential of mitophagy-inducing microflora metabolite,urolithin a for alzheimer's disease[J].Nutrients,13.

[37]Qin,X.,Li,H.,Zhao,H.,et al.(2024).Enhancing healthy aging with small molecules:Amitochondrial perspective[J].Medicinal research reviews,10.1002/med.22034.Advance online publication.

[38]马小燕,张长飞.(2020).一种尿石素A的合成方法[P]. CN111978282A.

[39]游贤会,赵甜甜.(2023).抗衰老药物尿石素a合成工艺优化研究[J].上海化工,48(1),11-14.

[40]Ito,H.,Iguchi,A.,Hatano,T.(2008).Identification of urinary and intestinal bacterial metabolites of ellagitannin geraniin in rats[J].Journal of agricultural and food chemistry,56(2),393-400.

[41]Sarkar,S.,Siddiqui,A.,Mazumder,S.,et al.(2015).Ellagic Acid,a Dietary Polyphenol,Inhibits Tautomerase Activity of Human Macrophage Migration Inhibitory Factor and Its Pro-inflammatory Responses in Human Peripheral Blood Mononuclear Cells[J].Journal of agricultural and food chemistry,63(20),4988-4998.

[42]Thasana,N..(2014).Cu(i)-mediated lactone formation in subcritical water:a benign synthesis of benzopyranones and urolithins a–c[J].Tetrahedron,45(44),9277-9283.

[43]Yin,P.,Zhang,J.,Yan,L.,Yang,L.,et al.(2017).Urolithin c,a gut metabolite of ellagic acid,induces apoptosis in pc12 cells through a mitochondria-mediated pathway[J].RSC Advances,7.

[44]Ross,P.J.,Goissis,M.D.,Martins,J.P.N.,Chitwood,J.L.,et al.(2023).Blastocyst cell number and allocation affect the developmental potential and transcriptome of bovine somatic cell nuclear transfer embryos[J].Stem cells and development.

[45]García-Villalba,R.,Giménez-Bastida,J.A.,Cortés-Martín,A.,et al.(2022).Urolithins:a Comprehensive Update on their Metabolism,Bioactivity,and Associated Gut Microbiota[J].Molecular nutrition &food research,66(21),e2101019.

[46]García-Villalba,R.,Giménez-Bastida,J.A.,Cortés-Martín,A.,et al.(2022).Urolithins:a Comprehensive Update on their Metabolism,Bioactivity,and Associated Gut Microbiota[J].Molecular nutrition &food research,66(21),e2101019.

[47]Francisco,A.,Tomás-Barberán,Antonio,González-Sarrías,et al.(2017).Urolithins,the rescue of "old" metabolites to understand a "new" concept:metabotypes as a nexus among phenolic metabolism,microbiota dysbiosis,and host health status[J].Molecular Nutrition &Food Research.

[48]Beltran,David,Romo-Vaquero.(2018).Ellagibacter isourolithinifaciens gen.nov. sp nov. a new member of the family eggerthellaceae,isolated from human gut[J].International Journal of Systematic and Evolutionary Microbiology.

[49]Gaya P,Peirotén Á,Medina M,et al.(2019) Bifidobacterium pseudocatenulatum INIA P815:The first bacterium able to produce urolithins A and B from ellagic acid[J].Journal of Functional Foods,2018,45:95-99.

[50]Liu,Q.,Liu,S.,Ye,Q.,(2018).(2022).A Novel Streptococcus thermophilus FUA329 Isolated from Human Breast Milk Capable of Producing Urolithin A from Ellagic Acid[J].Foods (Basel,Switzerland),11(20),32-80.

[51]房耀维,刘姝.(2023).嗜热链球菌FUA329及其发酵生产尿石素A的方法[P]. CN202211224627.6.

[52]Zhang,M.,Cui,S.,Mao,B.,et al.(2023).Urolithin A Produced by Novel Microbial Fermentation Possesses Anti-aging Effects by Improving Mitophagy and Reducing Reactive Oxygen Species in Caenorhabditis elegans[J].Journal of agricultural and food chemistry,71(16),6348-6357.

[53]唐鑫,张孟炜,毛丙永等.(2023).一株可产尿石素A抗衰老的植物乳杆菌及其应用[P].CN202211404309.8.

[54]房耀维,华梓延.(2023) 一种发酵粘液乳杆菌FUA033及其应用[P].

[55]刘姝,房耀维,杨光等.(2023).屎肠球菌FUA027及其产尿石素A的方法及应用[P].CN202211134634.7.

[56]Mi,H.,Liu,S.,Hai,Y.,et al.(2022).Lactococcus garvieae FUA009,a novel intestinal bacterium capable of producing the bioactive metabolite urolithin A from ellagic acid[J].Foods,11(17),2621.

[57]房耀维,刘姝,杨光等.(2023) 格氏乳球菌FUA009及其产尿石素A的方法[P].

[58]Cerón,José J.(2003).Repeated oral administration of high doses of the pomegranate ellagitannin punicalagin to rats for 37 days is not toxic[J].

[59]Azorín-Ortuo,María,Urbán,Cristina,et al.(2008).Safety evaluation of an oak-flavored milk powder containing ellagitannins upon oral administration in the rat[J].Journal of Agricultural &Food Chemistry,56(8),2857-2865.

[60]Espín,Juan Carlos,González-Barrio,et al.(2007).Iberian pig as a model to clarify obscure points in the bioavailability and metabolism of ellagitannins in humans[J].J Agric Food Chem,55(25),10476-10485.

[61]González-Barrio,Rocío,Truchado,P.,García-Villalba,et al.(2012).Metabolism of oak leaf ellagitannins and urolithin production in beef cattle[J].J Agric Food Chem,60(12),3068-3077.

[62]Cbegona,P.P.,Juan,C.E.,Francisco,A.,Tomaas,B..(2005).Identification of urolithin a as a metabolite produced by human colon microflora from ellagic acid and related compounds[J].

[63]Van,d.H.J.J.J.,De Vos,R.C.H.,Mihaleva,V.,et al.(2012).Structural elucidation and quantification of phenolic conjugates present in human urine after tea intake[J].Analytical Chemistry,84(16),7263.

[64]Mertenstalcott,S.U.,Jilmastohlawetz,P.,Rios,J .(2006).Absorption,metabolism,and antioxidant effects of pomegranate (punica granatum l.) polyphenols after ingestion of a standardized extract in healthy human volunteers[J].Journal of Agricultural &Food Chemistry,54(23),8956-61.

[65]Susanne M.Henning.(2013) Pomegranate juice alters the microbiota in breast milk and infant stool:a pilot study[J].Food &Function.

[66]Selma,M.V.,Beltrán,D.,Luna,M.C.,et al.(2017).Isolation of human intestinal bacteria capable of producing the bioactive metabolite isourolithin a from ellagic acid[J].Frontiers in microbiology,8,1521.

[67]Cerdá,B.,Periago,P.,Espín,J.C.,et al.(2005).Identification of urolithin A as a metabolite produced by human colon microflora from ellagic acid and related compounds[J].Journal of agricultural and food chemistry,53(14),5571-5576.

[68]Singh,A.,Andreux,P.,Blanco-Bose,W.,et al.(2017).Orally administered urolithin a is safe and modulates muscle and mitochondrial biomarkers in elderly[J].Innovation in Aging(suppl_1),suppl_1.

[69]Raimundo,A.,Ferreira,S.,Barberan,F.T.,et al.(2021).Urolithins:diet-derived bioavailable metabolites to tackle diabetes[J].Nutrients.

[70]Woldemariam,G.,Kyad,A.,Moore,S.,et al.(2020).尿素和相关杂质的 HPLC-UV 方法的开发和验证[J].PDA 药物科学与技术杂志,74（1）,2-14.

[71]Kalili,K.M.,de Villiers,A.(2011).Recent developments in the HPLC separation of phenolic compounds[J].Journal of separation science,34(8),854-876.

[72]Jiang,C.S.,Li,C.Y.,Gu,C.T.(2024).Bifidobacterium apis sp.nov.,isolated from the gut of honeybee (Apis mellifera)[J].International journal of systematic and evolutionary microbiology,74(4),10.1099/ijsem.0.006358.

[73]Corcoll,N.,Österlund,T.,Sinclair,L.,et al.(2017).Comparison of four DNA extraction methods for comprehensive assessment of 16S rRNA bacterial diversity in marine biofilms using high-throughput sequencing[J].FEMS microbiology letters,364(14),10.1093/femsle/fnx139.

[74]肖仔君,钟瑞敏,陈惠等.(2005).植物乳杆菌的生理功能与应用[J].中国食品添加剂,000(002),87-89.

[75]Seddik,H.A.,Bendali,F.,Gancel,F.,Fliss,I.,et al.(2017).Lactobacillus plantarum and Its Probiotic and Food Potentialities[P].Probiotics and antimicrobial proteins,9(2),111-122.

[76]Charteris,W.P.,Kelly,P.M.,Morelli,L.,et al.(1997).Selective detection,enumeration and identification of potentially probiotic Lactobacillus and Bifidobacterium species in mixed bacterial populations[J].International journal of food microbiology,35(1),1-27.

[77]Skåra,T.,Cappuyns,A.M.,van Derlinden,E.,et al.(2012).Growth kinetics of listeria isolated from salmon and salmon processing environment:single strains versus cocktails[J].Journal of food protection,75(7),1227-1235.

[78]Chillar,K.,Yin,Y.,Eriyagama,A.M.D.N.,et al.(2022).Determination of optical density (OD) of oligodeoxynucleotide from HPLC peak area[J].PeerJ analytical chemistry,4,e20.

[79]Mianzhi,Y.,Shah,N.P.(2017).Contemporary nucleic acid-based molecular techniques for detection,identification,and characterization of Bifidobacterium[J].Critical reviews in food science and nutrition,57(5),987-1016.

[80]Ryu,D.,Mouchiroud,L.,Andreux,et al.(2016).Urolithin A induces mitophagy and prolongs lifespan in C[J].elegans and increases muscle function in rodents.Nature medicine,22(8),879-888.

[81]Hsieh Y.(2008).HPLC-MS/MS in drug metabolism and pharmacokinetic screening[J].Expert opinion on drug metabolism &toxicology,4(1),93-101.

[82]Janda,J.M.,Abbott,S.L.(2007).16S rRNA gene sequencing for bacterial identification in the diagnostic laboratory:pluses,perils,and pitfalls[J].Journal of clinical microbiology,45(9),2761-2764.

[83]O'Farrell,K.A.,Janssen,P.H.(1999).Detection of Verrucomicrobia in a pasture soil by PCR-mediated amplification of 16S rRNA genes[J].Applied and environmental microbiology,65(9),4280-4284.

[84]Turroni,F.,van Sinderen,D.,Ventura,M.(2011).Genomics and ecological overview of the genus Bifidobacterium[J].International journal of food microbiology,149(1),37-44.

[85]Modesto,M.,Satti,M.,Watanabe,K.,et al.(2020).Phylogenetic characterization of two novel species of the genus Bifidobacterium:Bifidobacterium saimiriisciurei sp.nov.and Bifidobacterium platyrrhinorum sp.nov[J].Systematic and applied microbiology,43(5),1261-11.

[86]Van Wey,A.S.,Cookson,A.L.,Roy,N.C.,McNabb,et al.(2014).Monoculture parameters successfully predict coculture growth kinetics of Bacteroides thetaiotaomicron and two Bifidobacterium strains[J].International journal of food microbiology,191,172-181.

[87]Egli T.(2010).How to live at very low substrate concentration[J].Water research,44(17),4826–4837.

[88]方子季,许自明,朱秀美等.(2002).高效液相色谱内标法测定银杏叶制品中的总黄酮含量[J].解放军药学学报,18(1),3.

[89]白峰,徐国宾.(2020).反相高效液相色谱内标法同时测定血清肌酐和尿酸[J].中华医学检验杂志 1993年16卷2期,79-82页,ISTIC PKU CSCD CA.

[90]王天志,李永梅,王志霄.(2000).金银花中3种有机酸的反相高效液相色谱法定量分析[J].药物分析杂志,020(005),293-296

[91]Sanayhie,S.A.,&Sabatinos,S.A.(2025).Identifying Drug Sensitivities in Fission Yeast by Assessing Growth Kinetics[J].Methods in molecular biology (Clifton,N.J.),2862,321-331.

[92]Echegaray,N.,Yilmaz,B.,Sharma,H.,et al.(2023).A novel approach to Lactiplantibacillus plantarum:From probiotic properties to the omics insights[J].Microbiological research,268,127-289.

[93]肖仔君,钟瑞敏,陈惠音等.(2005).植物乳杆菌的生理功能与应用[J].中国食品添加剂,000(002),87-89.