硝尔库勒湖可培养放线菌多样性及其功能酶和抗细菌活性
|
摘要
【目的】认识和了解硝尔库勒湖可培养放线菌的多样性、功能酶和抗细菌活性特点,为今后的开发和利用奠定基础。【方法】应用可培养技术和基于16S r RNA基因序列的系统发育分析硝尔库勒盐湖沉积物中放线菌的多样性。常规方法检测样品成分因子,并筛选了嗜盐放线菌的蛋白酶、淀粉酶和酯酶活性;抑菌圈法检测放线菌新种的抗细菌活性。【结果】分离获得了51个OTUs,分属于24个不同的属,其中15个OTUs代表了放线菌新种;链霉菌属是优势菌属,占全部分离菌株数量的16.25%。硝尔库勒湖放线菌类群数量一定程度上受样品成分因子的协同影响。代表新种的菌株展示了良好的功能酶活性和抗细菌活性,其中代表链霉菌新种的菌株XHU5011不仅具有多种酶活性,而且具有强大的抗金黄葡萄球菌、耻垢分枝杆菌和荧光假单胞菌的能力,具有很好的开发潜能。【结论】硝尔库勒盐湖中存在丰富的可培养放线菌多样性,潜藏着大量的放线菌新资源,并且具有很好的功能酶和天然产物挖掘潜力。
[Objective] The study is aimed to reveal the diversity of culturable actinobacteria in Xiaoerkule salt lake, to lay a foundation for further development. [Methods] Actinobacterial diversity of Xiaoerkule lake was studied by culturable method and phylogenetic analysis based on 16 S rRNA gene sequences. Component factors of samples were tested by conventional methods. Activity for protease, amylase and asterase was tested by inoculating single colony method. Antibacterial activity of novel actinomycetes was detected by bacteriostasis ring method. [Results] In total 51 OTUs isolated from the salt lake belonged to 24 different genera, and 15 strains are novel actinomycetes. Streptomyces is predominated microorganisms for 16.25% of the total isolated amounts. Sample environmental factors affected the population of actinomycetes in this lake. Tested actinomycetes showed excellent functional enzyme activity and antibacterial activity. Strain XHU 5011, a novel actinomycete, showed great potential for anti-Staphylococcus aureus, anti-Mycobacterium smegmatis and anti-Pseudomonas fluoressens. [Conclusion] There is abundant actinobacterial diversity in the sediment of Xiaoerkule lake, and the result implies that there are large numbers of unknown actionobacterial groups here. These actinomycetes have excellent functional enzymes and provide the basis for further study on secondary metabolites.
[Objective] The study is aimed to reveal the diversity of culturable actinobacteria in Xiaoerkule salt lake, to lay a foundation for further development. [Methods] Actinobacterial diversity of Xiaoerkule lake was studied by culturable method and phylogenetic analysis based on 16 S rRNA gene sequences. Component factors of samples were tested by conventional methods. Activity for protease, amylase and asterase was tested by inoculating single colony method. Antibacterial activity of novel actinomycetes was detected by bacteriostasis ring method. [Results] In total 51 OTUs isolated from the salt lake belonged to 24 different genera, and 15 strains are novel actinomycetes. Streptomyces is predominated microorganisms for 16.25% of the total isolated amounts. Sample environmental factors affected the population of actinomycetes in this lake. Tested actinomycetes showed excellent functional enzyme activity and antibacterial activity. Strain XHU 5011, a novel actinomycete, showed great potential for anti-Staphylococcus aureus, anti-Mycobacterium smegmatis and anti-Pseudomonas fluoressens. [Conclusion] There is abundant actinobacterial diversity in the sediment of Xiaoerkule lake, and the result implies that there are large numbers of unknown actionobacterial groups here. These actinomycetes have excellent functional enzymes and provide the basis for further study on secondary metabolites.
引文
[1]关统伟.中国盐湖放线菌.北京:中国轻工业出版社,2017.
[2]Zhao XQ,Xu XN,Chen LY.Production of enzymes from marine actinobacteria.Advanced Food and Nutrition Research,2016,78(2):137-151.
[3]Reed KA,Manam RR,Mitchell SS,Xu JL,Teisan S,Chao TH,Deyanat-Yazdi G,Neuteboom STC,Lam KS,Potts BCM.Salinosporamides D-J from the marine actinomycete Salinispora tropica,Bromosalinosporamide,and Thioester derivatives are potent inhibitors of the 20S proteasome.Journal of Natural Products,2007,70(2):269-276.
[4]Yang XW,Zhang GY,Ying JX,Yang B,Zhou XF,Steinmetz A,Liu YH,Wang N.Isolation,characterization,and bioactivity evaluation of 3-((6-methylpyrazin-2-yl)methyl)-1H-indole,a new alkaloid from a deep-sea-derived actinomycete Serinicoccus profundi sp.nov.Marines Drugs,2013,11(1):33-39.
[5]Sharma AK,Singh SP.Effect of amino acids on the repression of alkaline protease synthesis in haloalkaliphilic Nocardiopsis dassonvillei.Biotechnology Report,2016,12(4):40-51.
[6]Guan TW,Wu JY,Zhi XY,Tang SK,Xu LH,Li WJ,Zhang LL.Actinobacterial diversity of a sediment sample from Xiaoerkule Lake.Acta Microbiologica Sinica,2008,48(7):851-856.(in Chinese)关统伟,吴晋元,职晓阳,唐蜀昆,徐丽华,李文均,张利莉.硝尔库勒湖沉积物中非培养放线菌多样性.微生物学报,2008,48(7):851-856.
[7]Mihai MM,Holban AM,Giurcaneanu C,Popa LG,Oanea RM.Microbial biofilms:impact on the pathogenesis of periodontitis,cystic fibrosis,chronic wounds and medical device-related infections.Current Topics Medicinal Chemistry,2015,15(16):1552-1576.
[8]Cui XL,Mao P,Zeng M,Li WJ,Zhang LP,Xu LH,Jiang CL.Streptimonospora salina gen.nov.,sp.nov.,a new member of family Nocardiopsaceae.International Journal of Systematic and Evolutionary Microbiology,2001,51(2):357-363.
[9]Yoon SH,Ha SM,Kwon S,Lim J,Kim Y,Seo H,Chun J.Introducing Ez Bio Cloud:A taxonomically united database of 16S r RNA gene sequences and whole-genome assemblies.International Journal of Systematic and Evolutionary Microbiology,2017,67(5):1613-1617.
[10]Legendre P,Gallagher ED.Ecologically meaningful transformations for ordination of species data.Oecologia,2001,129(2):271-280.
[11]Demain AL,Sanchez S.Microbial drug discovery:80 years of progress.Journal of Antibiotics,2009,62(3):5-16.
[12]Fenical W,Baden D,Burg M.Marinederived pharmaceuticals and related bioactive compounds//Fenical W.From Monsoons to Microbes:Understanding the Ocean’s Role in Human Health.National Academies Press,1999,6:71-86.
[13]Flaxman SM,Feder JL,Nosil P.Genetic hitchhiking and the dynamic buildup of genomic divergence during speciation with gene flow.Evolution,2013,67(9):2577-2591.
[14]Shafer ABA,Wolf JBW.Widespread evidence for incipient ecological speciation:a meta-analysis of isolation-by-ecology.Ecology Letters,2013,16(7):940-950.
[15]Shi YY,Li XZ,Zhang GM.Advances in microbial halophilic enzymes.Acta Microbiologica Sinica,2017,57(8):1180-1188.(in Chinese)石云云,李信志,张桂敏.微生物嗜盐酶的研究进展.微生物学报,2017,57(8):1180-1188.
[16]Hamedi J,Mohammadipanah F,Ventosa A.Systematic and biotechnological aspects of halophilic and halotolerant actinomycetes.Extremophiles,2013,17(1):1-13.
[17]Liu WH,Li MG,Li YQ,Zhao JY,Ding ZG,Yang PW,Cui XL,Wen ML.Cytotoxic metabolites of Streptimonospora salina.Chemistry of Natural Compounds,2008,44(4):503-505.
[18]Asolkar RN,Freel KC,Jensen PR,Fenical W,Kondratyuk TP,Park EJ,Pezzuto JM.Arenamides A-C,ytotoxic NFκBinhibitors from the marine actinomycete Salinispora arenicola.Journal of Natural Products,2009,72(3):396-402.
[19]Zhao LX,Huang SX,Tang SK,Jiang CL,Duan Y,Beutler JA,Henrich CJ,Mc Mahon JB,Schmid T,Blees JS,Colburn NH,Rajski SR,Shen B.Actinopolysporins A-C and tubercidin as a Pdcd4 stabilizer from the halophilic actinomycete Actinopolyspora erythraea YIM 90600.Journal of Natural Products,2011,74(9):1990-1995.
[20]Zhang QB,Li SM,Chen YC,Tian XP,Zhang HB,Zhang GT,Yiguang Zhu,Zhang S,Zhang WM,Zhang CS.New diketopiperazine derivatives from a deep-sea-derived Nocardiopsis alba CSIO 03039.The Journal of Antibiotics,2013,66(1):31-36.
[21]Chen CX,Song FH,Wang Q,Abdelmageed WM,Guo H,Fu CZ,Hou WY,Dai HQ,Liu XT,Yang N,Xie F,Yu K,Chen RX,Zhang LX.A marine-derived Streptomyces sp.MS449produces high yield of actinomycin X2 and actinomycin Dwith potent anti-tuberculosis activity.Applid Microbiology and Biotechnolog,2012,95(4):919-927.
[2]Zhao XQ,Xu XN,Chen LY.Production of enzymes from marine actinobacteria.Advanced Food and Nutrition Research,2016,78(2):137-151.
[3]Reed KA,Manam RR,Mitchell SS,Xu JL,Teisan S,Chao TH,Deyanat-Yazdi G,Neuteboom STC,Lam KS,Potts BCM.Salinosporamides D-J from the marine actinomycete Salinispora tropica,Bromosalinosporamide,and Thioester derivatives are potent inhibitors of the 20S proteasome.Journal of Natural Products,2007,70(2):269-276.
[4]Yang XW,Zhang GY,Ying JX,Yang B,Zhou XF,Steinmetz A,Liu YH,Wang N.Isolation,characterization,and bioactivity evaluation of 3-((6-methylpyrazin-2-yl)methyl)-1H-indole,a new alkaloid from a deep-sea-derived actinomycete Serinicoccus profundi sp.nov.Marines Drugs,2013,11(1):33-39.
[5]Sharma AK,Singh SP.Effect of amino acids on the repression of alkaline protease synthesis in haloalkaliphilic Nocardiopsis dassonvillei.Biotechnology Report,2016,12(4):40-51.
[6]Guan TW,Wu JY,Zhi XY,Tang SK,Xu LH,Li WJ,Zhang LL.Actinobacterial diversity of a sediment sample from Xiaoerkule Lake.Acta Microbiologica Sinica,2008,48(7):851-856.(in Chinese)关统伟,吴晋元,职晓阳,唐蜀昆,徐丽华,李文均,张利莉.硝尔库勒湖沉积物中非培养放线菌多样性.微生物学报,2008,48(7):851-856.
[7]Mihai MM,Holban AM,Giurcaneanu C,Popa LG,Oanea RM.Microbial biofilms:impact on the pathogenesis of periodontitis,cystic fibrosis,chronic wounds and medical device-related infections.Current Topics Medicinal Chemistry,2015,15(16):1552-1576.
[8]Cui XL,Mao P,Zeng M,Li WJ,Zhang LP,Xu LH,Jiang CL.Streptimonospora salina gen.nov.,sp.nov.,a new member of family Nocardiopsaceae.International Journal of Systematic and Evolutionary Microbiology,2001,51(2):357-363.
[9]Yoon SH,Ha SM,Kwon S,Lim J,Kim Y,Seo H,Chun J.Introducing Ez Bio Cloud:A taxonomically united database of 16S r RNA gene sequences and whole-genome assemblies.International Journal of Systematic and Evolutionary Microbiology,2017,67(5):1613-1617.
[10]Legendre P,Gallagher ED.Ecologically meaningful transformations for ordination of species data.Oecologia,2001,129(2):271-280.
[11]Demain AL,Sanchez S.Microbial drug discovery:80 years of progress.Journal of Antibiotics,2009,62(3):5-16.
[12]Fenical W,Baden D,Burg M.Marinederived pharmaceuticals and related bioactive compounds//Fenical W.From Monsoons to Microbes:Understanding the Ocean’s Role in Human Health.National Academies Press,1999,6:71-86.
[13]Flaxman SM,Feder JL,Nosil P.Genetic hitchhiking and the dynamic buildup of genomic divergence during speciation with gene flow.Evolution,2013,67(9):2577-2591.
[14]Shafer ABA,Wolf JBW.Widespread evidence for incipient ecological speciation:a meta-analysis of isolation-by-ecology.Ecology Letters,2013,16(7):940-950.
[15]Shi YY,Li XZ,Zhang GM.Advances in microbial halophilic enzymes.Acta Microbiologica Sinica,2017,57(8):1180-1188.(in Chinese)石云云,李信志,张桂敏.微生物嗜盐酶的研究进展.微生物学报,2017,57(8):1180-1188.
[16]Hamedi J,Mohammadipanah F,Ventosa A.Systematic and biotechnological aspects of halophilic and halotolerant actinomycetes.Extremophiles,2013,17(1):1-13.
[17]Liu WH,Li MG,Li YQ,Zhao JY,Ding ZG,Yang PW,Cui XL,Wen ML.Cytotoxic metabolites of Streptimonospora salina.Chemistry of Natural Compounds,2008,44(4):503-505.
[18]Asolkar RN,Freel KC,Jensen PR,Fenical W,Kondratyuk TP,Park EJ,Pezzuto JM.Arenamides A-C,ytotoxic NFκBinhibitors from the marine actinomycete Salinispora arenicola.Journal of Natural Products,2009,72(3):396-402.
[19]Zhao LX,Huang SX,Tang SK,Jiang CL,Duan Y,Beutler JA,Henrich CJ,Mc Mahon JB,Schmid T,Blees JS,Colburn NH,Rajski SR,Shen B.Actinopolysporins A-C and tubercidin as a Pdcd4 stabilizer from the halophilic actinomycete Actinopolyspora erythraea YIM 90600.Journal of Natural Products,2011,74(9):1990-1995.
[20]Zhang QB,Li SM,Chen YC,Tian XP,Zhang HB,Zhang GT,Yiguang Zhu,Zhang S,Zhang WM,Zhang CS.New diketopiperazine derivatives from a deep-sea-derived Nocardiopsis alba CSIO 03039.The Journal of Antibiotics,2013,66(1):31-36.
[21]Chen CX,Song FH,Wang Q,Abdelmageed WM,Guo H,Fu CZ,Hou WY,Dai HQ,Liu XT,Yang N,Xie F,Yu K,Chen RX,Zhang LX.A marine-derived Streptomyces sp.MS449produces high yield of actinomycin X2 and actinomycin Dwith potent anti-tuberculosis activity.Applid Microbiology and Biotechnolog,2012,95(4):919-927.