东海海域海洋微生物活性物质的研究
|
摘要
随着抗生素及其他抗菌药物的广泛应用,病原微生物的耐药问题日趋严重,如耐甲氧西林金黄色葡萄球菌(MRSA)的分离率不断升高,已成为感染的重要致病菌之一。因此,开发新型抗生素迫在眉睫。陆地微生物曾是寻找抗生素的主要来源,但近年来从陆地微生物中发现新抗生素的几率越来越低,人们必须寻找新的药源。海洋环境的特殊性使海洋微生物具有独特的遗传背景与代谢途径,能产生结构新颖、作用独特的代谢产物,成为筛选新型抗生素的重要来源。因此,海洋微生物以其代谢产物的化学多样性、独特的生理活性和潜在的药物开发前景引起了科学家的广泛关注。
对东海海域不同地区采集的不同样品(包括海洋沉积物和海洋动物)用平板稀释法分离海洋微生物,实验表明培养基的组成和培养条件对海洋微生物的生长及活性物质的产生都有很大的影响。从三个海底沉积物样品中分离细菌466株,真菌53株,放线菌8株；从深海鲨鱼鱼鳃和胃肠道部位分离得到可培养微生物36株,来自鱼鳃5株,来自胃肠道31株,其中真菌7株,占19%；放线菌1株,占3%；细菌28株,占78%。以枯草芽孢杆菌(Bacillus subtilis)、金黄色葡萄球菌(Staphylococcus aureus)、大肠杆菌(Escherichia coliC)、铜绿假单胞菌(Pseudomonas aeruginose)、产气肠杆菌(Enterobacter aerogenes)、白色念珠菌(Candida albicans)作为指示菌,对鲨鱼胃肠道中的菌株采用牛津杯法筛选产抗菌物质的微生物。
对分离自鲨鱼鳃及胃肠道的菌株进行形态、生理生化实验、16SrRNA分子鉴定和系统发育分析,为进一步研制海洋抗菌活性物质提供一些基本参数和活性菌株。
本研究以非核糖体多肽酶系作为研究对象,以NRPSs A结构域保守序列设计简并引物,对鲨鱼胃肠道细菌进行了NRPSs的筛选,希望能获得NRPSs基因片段,为今后开发海洋来源的非核糖体多肽类物质打下基础。
从样品中分离得到了一株具有抗菌活性高、抗菌谱广的细菌,并对菌株和其代谢抗菌产物进行了研究。研究证实,该菌株对革兰氏阴性菌、革兰氏阳性菌、真菌都有一定的拮抗作用,特别是对产气肠杆菌、金黄色葡萄球菌、白色念珠菌抗菌作用较强。对细菌Y9所产抗菌活性物质理化性质初步研究表明,该抗菌活性成分热稳定性较强,酸性、中性条件下稳定,碱性条件下不稳定,非蛋白类物质,易溶于甲醇、乙醇。
With the wide use of antibiotics and other antibacterial agents, bacteria resistant to many antibiotics is increasing seriously, for example, Methicillin-resistant Staphylococcus aureus (MRSA) which is resistant to most antibiotics is increasing rapidly and has become the vital pathogenic bacteria in the clinic. A renewed effort should be made to seek new antibiotics. Microorganism resources in land once were the main source for screening for antibiotic. However, with the research going on, it is becoming more and more dificult to find new antibiotics. It is necessary to seek for the new medicine source. Since the specific living environment, marine microorganisms have unique heredity background and metabolite pathways, and synthesize some distinct antibiotics in structures. Therefor it has significance for antibiotics screening from marine microorganism. And the marine microorganisms have attracted considerable attention because of chemical unique, rich physiological properties and thus potential as important drugs of their metabolites.
Many kinds of samples, in cluding sea mud and sea vertebrata were collected from East Sea from different region. It is greatly influenced by the composition of medium and culture condition on the production of the metabolites with antimicrobial activity and the growth of marine microorganism.446 strains marine bacteria,8 strains marine actinomycetes and 53 strains marine fungi were isolated from the sea mud. Total 36 culturable microorganisms,28 marine bacteria,1 strains marine actinomycetes and 7 strains marine fungi were isolated from the gills, stomachs or intestines of deep-sea shark,of which 5 strains were from gills and 31 strains were from stomachs or intestines.Among them,there were 7 strains of fungi (19%),1 strains of actinomycetes (3%) and 28 strains of bacteria (78%). Their antibiotic activities against six microorganisms (Bacillus subtilis、Staphylococcus aureus、Escherichia coli、Pseudomonas aeruginose、Enterobacter aerogenes、Candida albicans) were assayed Oxford Cup method.
The isolated strains from the gills, stomachs or intestines of deep-sea shark is identified through morphological, physiological and biochemical experiment,16S rRNA sequence analysis and phylogenetic analysis. It will provide some basic information and some bioactive strains for the discovery of the metabolites with antimicrobial activity from sea.
In this study, a pair of degenerate oligonucleotide primers, disigned for amplification A domain in NRPSs, had been employed to screen NRPSs gene fragments from the gills, stomachs or intestines of deep-sea shark. Our purpose was to develop a method to access NRPSs gene diversity in sea. This would open up the possibilities of the secondary metabolites and as the basis for a search for attractive antibiotic biosynthesis genes that could be used in the heterogenous expression and combinatorial biosynthesis. Furthermore, the amplified NRPSs gene fragments can also be used as homologous hybridization probes to detect the clones harbored NRPSs gene clusters.
One strain is screened from the gills,stomachs or intestines of deep-sea shark. The metabolites produced by the strain have high antimicrobial activity and broad antimicrobial spectrum. The strain and its metabolites are researched. The results show that the culture has a broad antimicrobial spectrum against some gram-negative and gram-positive bacteria, and moulds, especially against Enterobacter aerogenes, Staphylococcus aureus and Candida albicans. The physical and chemical properies of the metabolites with antimicrobial activity of Y9, the results showed it has good stability against temperature and acidity, but not alkali. The metabolites with antimicrobial activity are identified which is not a kind of protein, and it can be dissolved in methyl alcohol and ethanol.
对东海海域不同地区采集的不同样品(包括海洋沉积物和海洋动物)用平板稀释法分离海洋微生物,实验表明培养基的组成和培养条件对海洋微生物的生长及活性物质的产生都有很大的影响。从三个海底沉积物样品中分离细菌466株,真菌53株,放线菌8株；从深海鲨鱼鱼鳃和胃肠道部位分离得到可培养微生物36株,来自鱼鳃5株,来自胃肠道31株,其中真菌7株,占19%；放线菌1株,占3%；细菌28株,占78%。以枯草芽孢杆菌(Bacillus subtilis)、金黄色葡萄球菌(Staphylococcus aureus)、大肠杆菌(Escherichia coliC)、铜绿假单胞菌(Pseudomonas aeruginose)、产气肠杆菌(Enterobacter aerogenes)、白色念珠菌(Candida albicans)作为指示菌,对鲨鱼胃肠道中的菌株采用牛津杯法筛选产抗菌物质的微生物。
对分离自鲨鱼鳃及胃肠道的菌株进行形态、生理生化实验、16SrRNA分子鉴定和系统发育分析,为进一步研制海洋抗菌活性物质提供一些基本参数和活性菌株。
本研究以非核糖体多肽酶系作为研究对象,以NRPSs A结构域保守序列设计简并引物,对鲨鱼胃肠道细菌进行了NRPSs的筛选,希望能获得NRPSs基因片段,为今后开发海洋来源的非核糖体多肽类物质打下基础。
从样品中分离得到了一株具有抗菌活性高、抗菌谱广的细菌,并对菌株和其代谢抗菌产物进行了研究。研究证实,该菌株对革兰氏阴性菌、革兰氏阳性菌、真菌都有一定的拮抗作用,特别是对产气肠杆菌、金黄色葡萄球菌、白色念珠菌抗菌作用较强。对细菌Y9所产抗菌活性物质理化性质初步研究表明,该抗菌活性成分热稳定性较强,酸性、中性条件下稳定,碱性条件下不稳定,非蛋白类物质,易溶于甲醇、乙醇。
With the wide use of antibiotics and other antibacterial agents, bacteria resistant to many antibiotics is increasing seriously, for example, Methicillin-resistant Staphylococcus aureus (MRSA) which is resistant to most antibiotics is increasing rapidly and has become the vital pathogenic bacteria in the clinic. A renewed effort should be made to seek new antibiotics. Microorganism resources in land once were the main source for screening for antibiotic. However, with the research going on, it is becoming more and more dificult to find new antibiotics. It is necessary to seek for the new medicine source. Since the specific living environment, marine microorganisms have unique heredity background and metabolite pathways, and synthesize some distinct antibiotics in structures. Therefor it has significance for antibiotics screening from marine microorganism. And the marine microorganisms have attracted considerable attention because of chemical unique, rich physiological properties and thus potential as important drugs of their metabolites.
Many kinds of samples, in cluding sea mud and sea vertebrata were collected from East Sea from different region. It is greatly influenced by the composition of medium and culture condition on the production of the metabolites with antimicrobial activity and the growth of marine microorganism.446 strains marine bacteria,8 strains marine actinomycetes and 53 strains marine fungi were isolated from the sea mud. Total 36 culturable microorganisms,28 marine bacteria,1 strains marine actinomycetes and 7 strains marine fungi were isolated from the gills, stomachs or intestines of deep-sea shark,of which 5 strains were from gills and 31 strains were from stomachs or intestines.Among them,there were 7 strains of fungi (19%),1 strains of actinomycetes (3%) and 28 strains of bacteria (78%). Their antibiotic activities against six microorganisms (Bacillus subtilis、Staphylococcus aureus、Escherichia coli、Pseudomonas aeruginose、Enterobacter aerogenes、Candida albicans) were assayed Oxford Cup method.
The isolated strains from the gills, stomachs or intestines of deep-sea shark is identified through morphological, physiological and biochemical experiment,16S rRNA sequence analysis and phylogenetic analysis. It will provide some basic information and some bioactive strains for the discovery of the metabolites with antimicrobial activity from sea.
In this study, a pair of degenerate oligonucleotide primers, disigned for amplification A domain in NRPSs, had been employed to screen NRPSs gene fragments from the gills, stomachs or intestines of deep-sea shark. Our purpose was to develop a method to access NRPSs gene diversity in sea. This would open up the possibilities of the secondary metabolites and as the basis for a search for attractive antibiotic biosynthesis genes that could be used in the heterogenous expression and combinatorial biosynthesis. Furthermore, the amplified NRPSs gene fragments can also be used as homologous hybridization probes to detect the clones harbored NRPSs gene clusters.
One strain is screened from the gills,stomachs or intestines of deep-sea shark. The metabolites produced by the strain have high antimicrobial activity and broad antimicrobial spectrum. The strain and its metabolites are researched. The results show that the culture has a broad antimicrobial spectrum against some gram-negative and gram-positive bacteria, and moulds, especially against Enterobacter aerogenes, Staphylococcus aureus and Candida albicans. The physical and chemical properies of the metabolites with antimicrobial activity of Y9, the results showed it has good stability against temperature and acidity, but not alkali. The metabolites with antimicrobial activity are identified which is not a kind of protein, and it can be dissolved in methyl alcohol and ethanol.
引文
[1]Sherly AP. The bioprocess-technoiogical potential of the sea [J]. Biotechnoiogy, 1999,70:5-13.
[2]Pace NR. A molecular view of microbial diversity and the biosphere [J]. Science, 1997,276(5313):734-740.
[3]Wise MG, Mcarthur JV, Shimketa LJ. Bacterial diversity of a Carolina Bay as determined by 16S rRNA gene analysis:confirmation of novel taxa [J]. Appl Environ Microbiol,1997,63:1505-1514.
[4]Giovannoni SJ, Britschgi TB, Moyer CL, et al. Genetic diversity in Sargasso Sea bacterioplankton [J]. Nature,1990,345:60-63.
[5]刘雪莉,钱伯初.日本海洋天然活性物质研究简况[J].中国海洋药物,1997,61(1)：45-48.
[6]Panlilio AL. Methicillin-resistant Staphylococcus aureus in U. S. hospitals, 1975-1991 [J]. Infect Control Hosp Epidemiol,1992,13:582-586.
[7]王祥敏,李明,骆祝华,等.热带太平洋活性微生物菌株的筛选和鉴定[J].台湾海峡,2006，25(4)：465-472.
[8]Carte BK. Biomedical potential of marine natural products [J]. Bioscience,1996, 46(4):271.
[9]姜健,杨宝灵,等.海洋微生物生物活性物质的研究[J].云南大学学报,2004,26(6A)：91-95.
[10]Yarbrough GG, Taylor DP, Rowland RT, et al. Screening microbial metabolites for new drugs-theoretical and practical issues [J]. J.Antibiot, 1993,46:535-544.
[11]臧红梅,樊景凤,王彬,等.海洋微生物多样性的研究进展[J].海洋环境科学,2006,35(3)：96-100.
[12]Delong EF. Diversity of naturally occurring prokaryotes [A]. Microbial diversity in time and space [C]. New York:Plenum Press,1996.
[13]Istock CA, Bell JA, Ferguson N, et al. Bacteria diversity and evolution: Theoretical and practical perspectives [J]. J Ind Microbiol,1996, 17(3-4):137-150.
[14]Watve MG. Gangal RM. Problems in measuring bacterial diversity and a possible solution [J]. Applied and Environmental Microbiology,1996, 62(11):4299-4301.
[15]Okami Y. Marine microorganisms as a source of bioactive agents [J]. Microb Ecol,1986,12:65-78.
[16]李越中,陈琦.海洋微生物资源多样性[J].生物工程进展,1998，18(4)：34-40.
[17]相建海.海洋微生物[M].北京：科学出版社，2003.
[18]周德庆.微生物实验教程[M].北京：高等教育出版社,1993.
[19]史家栋,徐亚同,张圣章.环境微生物学[M].上海：华东师范大学出版社,1993.
[20]黄海燕.海洋微生物是药物和化学的重要资源[J].河北渔业.2001,1：7-9.
[21]展翔天,张春光,司玫.海洋微生物活性物质研究进展[J].中国海洋药物,2002,2：44-52.
[22]Proksch P, Edrada RA, Ebel R. Drugs from the seas-current status and microbiological implications [J]. Appl Microbiol Biotechnol,2002,59:125-128.
[23]谭仁祥.海洋微生物：新天然药物的重要源泉[J].中国天然药物,2006,4(1)：1-3.
[24]焦柄华,穆军,许强之,等.海洋微生物来源抗生素的研究[J].抗感染药学,2004,1(1)：1-4.
[25]Stritzke K, Schulz S, Laatsch H. Novel caprolactones from a marine streptomycete [J]. J Nat Prod,2004,67:395-398.
[26]晏仁义,陈若芸.天然产物中先导化合物的发展和优化[J].中国天然药物,2005,3(6)：322-324.
[27]李利菌,蔡慧农,苏文金.海洋微生物生物活性物质的研究[J].集美大学学报,2000,5(2)：80-82.
[28]汪开治.海洋生物药物研发现状综述[J].生物技术通报,2006，4：133-138.
[29]刘秀梅,邵守峰,薛丰松,等.自腹泻病人、海水、海产品中分离出麦氏弧菌及其毒素原性的研究[J].中国卫生检验杂志,1998，8(6)：359～360.
[30]马海萍,梅雪婷,等.国外海洋抗菌活性物质的研究进展[J].中国海洋药物杂志,2006,25(3)：46-49.
[31]Alphonse K. Secondary metabdites from marine microorganisms. [J] An Acad Bras Cienc,2002,74(1):151-170.
[32]Faulkner DJ. Marine natural products [J]. Nat Prod Rep,1998,15(2):113-189.
[33]Kelecorn A. Chemistry of marine natural products:yesterday,today and tomorrow [J]. An Acad Bras Cienc,1999,71:249—263.
[34]Hyde KD, Jones EB. Methods for the study of marine fungi from the mangroves [J]. Biodivesity and Conservation,1998,7:1147-1165.
[35]林永成,等.海洋生物化合物手册[M],北京：化学工业出版社,2006.
[36]Afiyatullov SS, Kuznetsova TA, Isakov VV, et al. New diterpenic altrosides of the fungus Acremonium stria tisporum isolated from a sea cucumber [J]. J Nat Prod,2000,63:848-852.
[37]Maneekarn C, Prasat K, Masahiko I, et al. Unique isocoumarin glucosides from marine fungus Halorosellinia [J]. J Chem Soc Pekin Ttans,2002,1:2473-2478.
[38]Belofsky GN, Anguera M, Jensen PR, et al. Oxepinamides A-C and Fumiquinazolines H-I:Bioactive metabolites from a marine isolate of a fungus of the genus Acremonium [J]. J Chem Eur,2000,6:1355-1357.
[39]岳家兴.海洋微生物与抗肿瘤活性物质[J].生命科学仪器,2006,4(10)：19-25.
[40]Pietra F. Secondary metabolites from marine microorganisms:bacteria, protozoa, algae and fungi achievements and prospects [J]. Nat Prod Rep,1997, 14(5):453-464.
[41]Attaway DH, Zaborsky OR. Marine biotechnology, pharmaceutical and bioactive natural products [M]. New York:Plenum Press,1993.
[42]Carte BK. Biomedical potential of marine natural products [J]. Bioscience,1996, 46(4):271.
[43]李根,陈瑞川,林昱.具抗肿瘤活性的海洋微生物菌株的初步筛选[J].台湾海峡,2002,21(1)：18-22.
[44]Flam F. Chemical prospectors scour the seas for promising drugs [J]. Science, 1994,266:1324-1325.
[45]Cui CB, Kakeya H, Osada H. Novel mammalian cell cycle inhibitors, spirotryprostatins A and B, produced by Aspergillus fumigatus, which G2/M phase [J], Tetrahedron,1996,52:12651-12658.
[46]Amagata T, Usami Y, Minoura K, et al. Cytotoxic subustances produced by a fungal strain from a sponge [J]. J Antibiot,1998,51(1):33-36.
[47]Belofsky GN, Jensen PR, Renner MK. New cytotoxic sesquiterpenoid nitrobenzoyl esters from a marine isolate of the fungus Aspergillus versicolor [J]. Tetrahedron,1998, 54:1715-1718.
[48]Fenical W. Chemical studies of marine bacteria: developing a new resource [J]. Chem Rev,1993,93:1673-1683.
[49]Renner MK, Jensen PR, Fenical W. Neomangicols:Structures and absolute stereochemistries of unprecedented halogenated sesterterpens from a marine fungus of the genus Fusatium. [J]. J Org Chem,1998,63:8346-8351..
[50]Xu XM, Guzman FSD, Gloer JB. Stachybotrins A and B:novel bioactive metabolites from a brackish water isolate of the fungus Stachybitrys sp. [J]. J Org Chem,1992,57:6700-6704.
[51]Ogita T, Hayashi T, Sato A. Furutani Feb. Japanese patent 62040292.1987.
[52]Carmichael WW. Cyanbobacteria secondary metabolites-the Cyanotoxins [J]. Journal of Applied Bactriology,1992,72:45-49.
[53]Yu CM, Curtis MJ, Walter JA [J]. J Antibiot,1996,49:395.
[54]Osterhage C, Kaminsky R, Konig GM. Ascosalipyrrolidinone A, an antimicrobial alkaloid, from the obligate marine fungus Ascochyta salicorniae [J]. J Org Chem, 2005,65:6412-6415.
[55]Alivik A, Casey A, Nair BG [J] J Antibiot,1998,51:515-519.
[56]王唯玮.海产油脂中心血管活性物质——高度不饱和脂肪酸的提取分离技术和应用[J].中国海洋药物,1989,3：30-43.
[57]李光友.海洋生物活性物质研究开发进展[C].国家海洋局海洋生物活性物质重点实验室及山东海洋生化工程技术研究中心研究年报论文集,青岛：青岛海洋大学出版社,2000.
[58]刘晶晶,曾全震,曾江宁,等.海洋微生物活性物质的研究进展[J].海洋学研究,2007,25(1)：55-65.
[59]李环,韦萍,欧阳平凯.嗜盐隐杆藻胞外多糖的分离、纯化及理化性质[J].天然产物研究与开发,2000,12(2)：52-56.
[60]Lin WH, Grabley S, et al. New cyclopentenone derivatives from an endophytic Streptomyces sp. isolated from the Mangrove Plant Aegiceras comiculatum [J]. Chin J Nat Med,2006,5:20-23.
[61]Nogva HK, Kristine KR, Naterstad K, et al. Application of 5'-nuclease PCR for quantitative detection of Listeria monocytogenes in pure cultures, water, skim milk, and unpasteurized whole milk [J|. Appl Environ Microbiol,2006, 66:4266-4271.
[62]Sykes PJ, Neoh SH, Brisco MJ, et al. Quantification of targets for PCR by use of limiting dilution [J]. Bio Techniques,1992,13:444-449.
[63]Dunbar J, Takala S, Barns MS, et al. Levels of bacterial community diversity in four arid soils compared by cultivation and 16S rRNA gene cloning [J]. Appl Environ Microbiol,1999,65:1662-1669.
[64]Finical W. Chemical studies of marine bacteria:Developing a new resource [J]. Chem Rev,1993,93:1673-1683.
[65]Gonzdez JM, Simo R, Massana R, et al. Bacterial community structure associated with a dmieyhyl sulfoniopropionate produing north Atlantic algal bloom [J]. Appl Environ Microbiol,2000,66:4237-4246.
[66]De Giaxa. Uber das Berhalten einiger pathogener Mikroorganismen in Meerwasser [J]. Zentralblatt fur hygiene und infectionkranheiten,1989, 6:162-225.
[67]Abraham EP, Loder PB. Cephalosporin C, in Cephalosporins and Penicillins:Chemistry and Biology [M]. New York:Academic press,1972.
[68]Burkholder PR, Pfister RM, Leitz FH. Production of a pyrrole antibiotic by a marine bacterium [J]. Appl Microbial,1966,14:649-653.
[69]Okami Y. Antibiotics from marine microorganisms with reference to plasmid involvement [J]. J Nat Prod,1979,42(6):589-595.
[70]Fenical W, Jensen PR, Mincer TJ. Marine actinomycete diversity and natural product discovery [J]. Antonie Van Leeuwenhoek,2005,87(1):43-48.
[71]郭跃伟.欧洲海洋药物的研究现状及对我国海洋药物研究的启示[J].医药经济信息,2000,20：22.
[72]穆军.海洋微生物抗生素国内外研究进展及方法学分析[D].上海：第二军医大学,2003：6.
[73]汪天虹,肖天,朱汇源,等.海洋丝状真菌生物活性物质研究进展[J].MarineSciences,2001,25(6)：25-27.
[74]Kelecom A. Secondary metabolites from marine microorganisms [J]. An Acad Bras Cienc,2002,74(1):151-170.
[75]Giovannoni SJ, Britschgi TB, Moyer CL, et al. Genetic diversity in Sargasso sea bacteriaplankton [J]. Nature,1990,345:60-63.
[76]Barns SM, Fundyha RE, Jeffeies MW, et al. Remarkable archaeal diversity detected in a Yellowstone National Park hot spring environment [J]. Proc Natl Acad Sci,1994,91:1609-1613.
[77]Torsvik V, Goksoyr J, Daae FL. High diversity in DNA of soil bacteria [J]. Appli Environ Microbiol,1990,56:782-787.
[78]Amans RI, Luding E, Schleifer KH. Phylogenetic identifiction and in situ detection of individual microbial cells without cultivation [J]. Microbial,1995, 66:4547-4554.
[79]Pace NR. A molecular viewer of microbial diversity and the biosphere [J]. Science,1997,276:734-740.
[80]Bach HJ, Errampalli D, Leung KT, et al. Specific detection of the gene for the extracellular neutral protease of Bacillus cereus by PCR and blot hybridization [J]. Appl Environ Microbiol,1999,65:3226-3228.
[81]Suzuki MT, Rappe MS, Haimberger ZW, et al. Bacterial diversity among small-subunit rRNA gene clones and cellular isolates from the same seawaters ample [J]. Applied and Environmental Microbiology,1997,63(3):983-989.
[82]Dai X, Zhou H, Cai CH, et al. A preliminary study on the novel taxa of bacteria in marine sediments [J]. Acta Scientiarum Naturalium Universitatis Suntvatseni, 2001,40(6):51-54.
[83]Dai X, Zhou H, Cai CH, et al. A preliminary study on the diversity of bacteria in the South China Sea Sedimentts as determined by 16S rRNA gene analysis [J]. Progress of Natural Science,2002,12(5):479-484.
[84]Jutta K, Oliver P, Martin HS, et al. Sulfate-reducing bacterial community response to carbon source amendments in contaminated aquifer microcosms [J]. FEMSMicrobiologv Ecologv,2002,42:109-118.
[85]Rasmussen LD, Ekelund F, Hansen LH. Group-specific PCR primers to amplify 24s a-subunit rRNA genes from Kinetoplastida (Protozoa) used in denaturing gradient gel electrophoresis [J]. Microb Ecol,2001,42:109-115.
[86]Beja 0, Sazuki MT, Heideberg JE, et al. Unsuspected diversity among marine aerobic anoxygenic phototrophs [J]. Nature,2002,415(6287):630-633.
[87]杨官品,茅云翔.环境细菌宏基因组研究及海洋细菌生物活性物质BAC文库筛选[J].青岛海洋大学学报,2001，31：718-722.
[88]Rondon MR, August PR, Bettermann AD, et al. Cloning the soil metagenome:a strategy for accessing the genetic and functional diversity of uncultured microorganisms [J]. Appl Environ Microbiol,2000,66:2541-2547.
[89]周世水,丁金国,姚汝华.海洋微生物药物的开发和应用[J].工业微生物,2002,2
[90]李渤,王乃力,姚新生,胡江春,王书锦.海洋微生物中具有引起稻瘟霉孢子形态变异活性的菌株筛选.沈阳药科大学学报,2002，4
[91]刘志鸿,程力,牟海津.海洋微生物活性物质的研究概况.中国水产科学,1999,4
[92]孙谧,洪义国,李勃生,修朝阳.海洋微生物低温酶特性及其在工业中的潜在用途.海洋水产研究,2002，3
[93]安鑫龙,周启星,刑光敏.海洋微生物在海洋污染治理中的应用现状.水产科学,2006,2
[94]Deming JW. Deep ocean environmental biotechnology [J]. Curr Opin Biotechnol, 1998,9:283-287.
[95]姜英,王秉瑞.抗生素耐药性机理研究进展[J].微生物学免疫学进展,2003,31(3)：74-76.
[96]Knight V, Sanglier JJ, Ditullio D, et al. Diversifying microbial natural products for drug discovery [J]. Appl Microbiol Biotechnol,2003,62(5-6):1-28.
[97]Simidu U, Lee WJ, Kogure K. Comparison of different techniques for determing plate counts of marine bacteria [J]. Bull Jpn Soc Sci Fish,1983,49:1199-1203.
[98]Richard EH, James AZ, Jeffrey DG, et al. Rapid method to estimate the presence of secondary metabolites in microbial extracts [J]. Appl Environ Microbial, 2001, 67(1):371-376.
[99]Amann R, Ludwig W. Ribosomal RNA-targeted nucleic acid probes for studies in microbial ecology [J]. FEMS Microbiol Rev,2000,24(5):555-565.
[100]林敏,宁喜斌.具抑菌活性海洋微生物的筛选[J].微生物学杂志,2005,(5)：23-25.
[101]Acar JF. The disc susceptibility test [Z]. In:Lorian V(ed) Antibiotics in laboratory medicine,1980.
[102]罗文新,陈晓佳,黄耀坚,等.来自海洋的链霉菌抑菌活性与其培养条件的关系[J].微生物学通报,1998,25(6)：325-332.
[103]Lee A, Langer R. Shark cartilage contains inhibitors of tumor angiogenesis [J]. Science,1983,221(4616):1185-1187.
[104]陈建鹤,焦炳华,缪为民,等.中国姥鲨软骨源新生血管抑制因子(Sp8)的分离纯化及生物学活性[J].第二军医大学学报,2000，21(2)：107-110.
[105]Damiani G, Amedeo P, Bandi C, et al. Bacteria indentification by PCR based techniques [M]. Microbial Genome Methods, CRC Press, 1996.
[106]O'Sullivan LA. Fuller KE. Thomas EM, et al. Distribution and culturability of the uncultivated "AGG58 cluster" of the Bacteriodetes phylum in aquatic environments [J]. FEMS Microbiol Ecol,2004, 47:359-370.
[107]Fry NK, Warwick S, Saunders NA, et al. J Gen Microbiol, 1991, 137:1215-1222.
[108]Devereux R, He SH, Doyle CL, et al. J Bacteriol,1990,172:3609-3619.
[109]Woese CR. Bacterial evolution [J]. Microbiol Rev,1987,51:221-271.
[110]Pace NR, Stahl DA, Lane DL, et al. The analysis of natural microbial populations by rRNA suquences [J]. Adv Microbiol Ecol,1986,9:1-55.
[111]vov Dohren H, Dieckmann R, et al. The nonribosomal code [J]. Chem Biol, 1999,6:273-279.
[112]明镇寰,潘建伟,朱睦元.非核糖体多肽合成酶进展[J].生物化学与生物物理进展,2002,29(5)：667-669.
[113]Stachelhaus T, Mooty HD, Marahiel MA. The specificity-conferring code of adenylation domains in nonribosomal peptide synthetases [J]. Chem Biol,1999, 6:493-505.
[114]Cane DE, Walsh CT. The parallel and convergent universes of polyketide synthases and nonribosomal peptide synthetases [J]. Chem Biol,1999, 6:319-325.
[115]Lambalot RH, Walsh CT. A new enzyme superfamily-the phosphopantetheinyl transferases [J]. Chem Biol,1996,3:923-936.
[116]Grunewald J, Marahiel MA. Chemoenzymatic and template-directed synthesis of bioactive macrocyclic peptides [J]. Microbiol Mol Biol Rev,2006,70(1):121-146.
[117]Stachelhaus T, Mooty HD, Bergendahl V, et al. Pepetide bond formation in nonribosomal peptide biosynthesis [J]. J Biol Chem,1998,272:22773-22781.
[118]Mootz HD, Schwarzer D, Marahiel MA. Ways of assembling complex natural products on modular nonribosomal peptide synthetases [J]. Chem Bio Chem, 2002,3(6):490-504.
[119]Becker JE, Moore R, Moore BS. Cloning, sequencing, and biochemical characterization of the nostocyclopeptide biosynthetic gene cluster.molecular basis for imine macrocyclization [J]. Gene,2004,325:35-42.
[120]Cramer RA Jr, Gamcsik MP, Brooking RM, et al. Disruption of a nonribosomal peptide synthetase in Aspergillus fumigatus eliminates gliotoxin production [J]. Eukaryot Cell,2006,5(6):972-801.
[121]Sharma A, Johri BN. Growth promotinginfluence of siderophore-producing Pseudomonas strains GRP3A and PRS9 in maize (Zea mays L.) under iron limiting conditions [J]. Microbiol Res,2003,158:243-248.
[122]Picosi S, Valladares A, Flores E, et al. Nitrogen-reulated genes for the metabolism of cyanophycin, a bacterial nitrogen reserve polymer [J]. J Bio Chem,2004,279(12):11582-11592.
[123]Liu F, Garneau S, Walsh CT. Hybrid nonribosomal peptide-polyketide interfaces in epothilone biosynthesis:minimal requirements at N and C termini of EpoB for elongation [J]. Chem Bio,2004,11:1533-1542.
[124]Miao V, Coeffet-Le Gal MF, Brian P.. et al. Daptomycin biosynthesis in Streptomyces roseosporus.cloning and analysis of the gene cluster and revision of peptide stereochemistry [J]. Microbiology,2005,151:1507-1523.
[125]Schneider A, Stachelhaus T, Marahiel MA. Targeted alteration of the substrate specificity of peptide synthetases by rationalmodule swapping [J]. Mol Gen Genet,1998,257:308-318.
[126]diDuilio M. The origin of peptide synthesis:On some molecular fossils identified through comparison of protein sequences [J]. Biosystems,1996,
39:169.
[127]Xu HS, Roberts N, Singleton FL, et al. Survival and viability of nonculturable Escherichia coli and Vibrio cholerae in the estuarine and marine environment [J]. Microbial Ecology,1982,8:313-323.
[2]Pace NR. A molecular view of microbial diversity and the biosphere [J]. Science, 1997,276(5313):734-740.
[3]Wise MG, Mcarthur JV, Shimketa LJ. Bacterial diversity of a Carolina Bay as determined by 16S rRNA gene analysis:confirmation of novel taxa [J]. Appl Environ Microbiol,1997,63:1505-1514.
[4]Giovannoni SJ, Britschgi TB, Moyer CL, et al. Genetic diversity in Sargasso Sea bacterioplankton [J]. Nature,1990,345:60-63.
[5]刘雪莉,钱伯初.日本海洋天然活性物质研究简况[J].中国海洋药物,1997,61(1)：45-48.
[6]Panlilio AL. Methicillin-resistant Staphylococcus aureus in U. S. hospitals, 1975-1991 [J]. Infect Control Hosp Epidemiol,1992,13:582-586.
[7]王祥敏,李明,骆祝华,等.热带太平洋活性微生物菌株的筛选和鉴定[J].台湾海峡,2006，25(4)：465-472.
[8]Carte BK. Biomedical potential of marine natural products [J]. Bioscience,1996, 46(4):271.
[9]姜健,杨宝灵,等.海洋微生物生物活性物质的研究[J].云南大学学报,2004,26(6A)：91-95.
[10]Yarbrough GG, Taylor DP, Rowland RT, et al. Screening microbial metabolites for new drugs-theoretical and practical issues [J]. J.Antibiot, 1993,46:535-544.
[11]臧红梅,樊景凤,王彬,等.海洋微生物多样性的研究进展[J].海洋环境科学,2006,35(3)：96-100.
[12]Delong EF. Diversity of naturally occurring prokaryotes [A]. Microbial diversity in time and space [C]. New York:Plenum Press,1996.
[13]Istock CA, Bell JA, Ferguson N, et al. Bacteria diversity and evolution: Theoretical and practical perspectives [J]. J Ind Microbiol,1996, 17(3-4):137-150.
[14]Watve MG. Gangal RM. Problems in measuring bacterial diversity and a possible solution [J]. Applied and Environmental Microbiology,1996, 62(11):4299-4301.
[15]Okami Y. Marine microorganisms as a source of bioactive agents [J]. Microb Ecol,1986,12:65-78.
[16]李越中,陈琦.海洋微生物资源多样性[J].生物工程进展,1998，18(4)：34-40.
[17]相建海.海洋微生物[M].北京：科学出版社，2003.
[18]周德庆.微生物实验教程[M].北京：高等教育出版社,1993.
[19]史家栋,徐亚同,张圣章.环境微生物学[M].上海：华东师范大学出版社,1993.
[20]黄海燕.海洋微生物是药物和化学的重要资源[J].河北渔业.2001,1：7-9.
[21]展翔天,张春光,司玫.海洋微生物活性物质研究进展[J].中国海洋药物,2002,2：44-52.
[22]Proksch P, Edrada RA, Ebel R. Drugs from the seas-current status and microbiological implications [J]. Appl Microbiol Biotechnol,2002,59:125-128.
[23]谭仁祥.海洋微生物：新天然药物的重要源泉[J].中国天然药物,2006,4(1)：1-3.
[24]焦柄华,穆军,许强之,等.海洋微生物来源抗生素的研究[J].抗感染药学,2004,1(1)：1-4.
[25]Stritzke K, Schulz S, Laatsch H. Novel caprolactones from a marine streptomycete [J]. J Nat Prod,2004,67:395-398.
[26]晏仁义,陈若芸.天然产物中先导化合物的发展和优化[J].中国天然药物,2005,3(6)：322-324.
[27]李利菌,蔡慧农,苏文金.海洋微生物生物活性物质的研究[J].集美大学学报,2000,5(2)：80-82.
[28]汪开治.海洋生物药物研发现状综述[J].生物技术通报,2006，4：133-138.
[29]刘秀梅,邵守峰,薛丰松,等.自腹泻病人、海水、海产品中分离出麦氏弧菌及其毒素原性的研究[J].中国卫生检验杂志,1998，8(6)：359～360.
[30]马海萍,梅雪婷,等.国外海洋抗菌活性物质的研究进展[J].中国海洋药物杂志,2006,25(3)：46-49.
[31]Alphonse K. Secondary metabdites from marine microorganisms. [J] An Acad Bras Cienc,2002,74(1):151-170.
[32]Faulkner DJ. Marine natural products [J]. Nat Prod Rep,1998,15(2):113-189.
[33]Kelecorn A. Chemistry of marine natural products:yesterday,today and tomorrow [J]. An Acad Bras Cienc,1999,71:249—263.
[34]Hyde KD, Jones EB. Methods for the study of marine fungi from the mangroves [J]. Biodivesity and Conservation,1998,7:1147-1165.
[35]林永成,等.海洋生物化合物手册[M],北京：化学工业出版社,2006.
[36]Afiyatullov SS, Kuznetsova TA, Isakov VV, et al. New diterpenic altrosides of the fungus Acremonium stria tisporum isolated from a sea cucumber [J]. J Nat Prod,2000,63:848-852.
[37]Maneekarn C, Prasat K, Masahiko I, et al. Unique isocoumarin glucosides from marine fungus Halorosellinia [J]. J Chem Soc Pekin Ttans,2002,1:2473-2478.
[38]Belofsky GN, Anguera M, Jensen PR, et al. Oxepinamides A-C and Fumiquinazolines H-I:Bioactive metabolites from a marine isolate of a fungus of the genus Acremonium [J]. J Chem Eur,2000,6:1355-1357.
[39]岳家兴.海洋微生物与抗肿瘤活性物质[J].生命科学仪器,2006,4(10)：19-25.
[40]Pietra F. Secondary metabolites from marine microorganisms:bacteria, protozoa, algae and fungi achievements and prospects [J]. Nat Prod Rep,1997, 14(5):453-464.
[41]Attaway DH, Zaborsky OR. Marine biotechnology, pharmaceutical and bioactive natural products [M]. New York:Plenum Press,1993.
[42]Carte BK. Biomedical potential of marine natural products [J]. Bioscience,1996, 46(4):271.
[43]李根,陈瑞川,林昱.具抗肿瘤活性的海洋微生物菌株的初步筛选[J].台湾海峡,2002,21(1)：18-22.
[44]Flam F. Chemical prospectors scour the seas for promising drugs [J]. Science, 1994,266:1324-1325.
[45]Cui CB, Kakeya H, Osada H. Novel mammalian cell cycle inhibitors, spirotryprostatins A and B, produced by Aspergillus fumigatus, which G2/M phase [J], Tetrahedron,1996,52:12651-12658.
[46]Amagata T, Usami Y, Minoura K, et al. Cytotoxic subustances produced by a fungal strain from a sponge [J]. J Antibiot,1998,51(1):33-36.
[47]Belofsky GN, Jensen PR, Renner MK. New cytotoxic sesquiterpenoid nitrobenzoyl esters from a marine isolate of the fungus Aspergillus versicolor [J]. Tetrahedron,1998, 54:1715-1718.
[48]Fenical W. Chemical studies of marine bacteria: developing a new resource [J]. Chem Rev,1993,93:1673-1683.
[49]Renner MK, Jensen PR, Fenical W. Neomangicols:Structures and absolute stereochemistries of unprecedented halogenated sesterterpens from a marine fungus of the genus Fusatium. [J]. J Org Chem,1998,63:8346-8351..
[50]Xu XM, Guzman FSD, Gloer JB. Stachybotrins A and B:novel bioactive metabolites from a brackish water isolate of the fungus Stachybitrys sp. [J]. J Org Chem,1992,57:6700-6704.
[51]Ogita T, Hayashi T, Sato A. Furutani Feb. Japanese patent 62040292.1987.
[52]Carmichael WW. Cyanbobacteria secondary metabolites-the Cyanotoxins [J]. Journal of Applied Bactriology,1992,72:45-49.
[53]Yu CM, Curtis MJ, Walter JA [J]. J Antibiot,1996,49:395.
[54]Osterhage C, Kaminsky R, Konig GM. Ascosalipyrrolidinone A, an antimicrobial alkaloid, from the obligate marine fungus Ascochyta salicorniae [J]. J Org Chem, 2005,65:6412-6415.
[55]Alivik A, Casey A, Nair BG [J] J Antibiot,1998,51:515-519.
[56]王唯玮.海产油脂中心血管活性物质——高度不饱和脂肪酸的提取分离技术和应用[J].中国海洋药物,1989,3：30-43.
[57]李光友.海洋生物活性物质研究开发进展[C].国家海洋局海洋生物活性物质重点实验室及山东海洋生化工程技术研究中心研究年报论文集,青岛：青岛海洋大学出版社,2000.
[58]刘晶晶,曾全震,曾江宁,等.海洋微生物活性物质的研究进展[J].海洋学研究,2007,25(1)：55-65.
[59]李环,韦萍,欧阳平凯.嗜盐隐杆藻胞外多糖的分离、纯化及理化性质[J].天然产物研究与开发,2000,12(2)：52-56.
[60]Lin WH, Grabley S, et al. New cyclopentenone derivatives from an endophytic Streptomyces sp. isolated from the Mangrove Plant Aegiceras comiculatum [J]. Chin J Nat Med,2006,5:20-23.
[61]Nogva HK, Kristine KR, Naterstad K, et al. Application of 5'-nuclease PCR for quantitative detection of Listeria monocytogenes in pure cultures, water, skim milk, and unpasteurized whole milk [J|. Appl Environ Microbiol,2006, 66:4266-4271.
[62]Sykes PJ, Neoh SH, Brisco MJ, et al. Quantification of targets for PCR by use of limiting dilution [J]. Bio Techniques,1992,13:444-449.
[63]Dunbar J, Takala S, Barns MS, et al. Levels of bacterial community diversity in four arid soils compared by cultivation and 16S rRNA gene cloning [J]. Appl Environ Microbiol,1999,65:1662-1669.
[64]Finical W. Chemical studies of marine bacteria:Developing a new resource [J]. Chem Rev,1993,93:1673-1683.
[65]Gonzdez JM, Simo R, Massana R, et al. Bacterial community structure associated with a dmieyhyl sulfoniopropionate produing north Atlantic algal bloom [J]. Appl Environ Microbiol,2000,66:4237-4246.
[66]De Giaxa. Uber das Berhalten einiger pathogener Mikroorganismen in Meerwasser [J]. Zentralblatt fur hygiene und infectionkranheiten,1989, 6:162-225.
[67]Abraham EP, Loder PB. Cephalosporin C, in Cephalosporins and Penicillins:Chemistry and Biology [M]. New York:Academic press,1972.
[68]Burkholder PR, Pfister RM, Leitz FH. Production of a pyrrole antibiotic by a marine bacterium [J]. Appl Microbial,1966,14:649-653.
[69]Okami Y. Antibiotics from marine microorganisms with reference to plasmid involvement [J]. J Nat Prod,1979,42(6):589-595.
[70]Fenical W, Jensen PR, Mincer TJ. Marine actinomycete diversity and natural product discovery [J]. Antonie Van Leeuwenhoek,2005,87(1):43-48.
[71]郭跃伟.欧洲海洋药物的研究现状及对我国海洋药物研究的启示[J].医药经济信息,2000,20：22.
[72]穆军.海洋微生物抗生素国内外研究进展及方法学分析[D].上海：第二军医大学,2003：6.
[73]汪天虹,肖天,朱汇源,等.海洋丝状真菌生物活性物质研究进展[J].MarineSciences,2001,25(6)：25-27.
[74]Kelecom A. Secondary metabolites from marine microorganisms [J]. An Acad Bras Cienc,2002,74(1):151-170.
[75]Giovannoni SJ, Britschgi TB, Moyer CL, et al. Genetic diversity in Sargasso sea bacteriaplankton [J]. Nature,1990,345:60-63.
[76]Barns SM, Fundyha RE, Jeffeies MW, et al. Remarkable archaeal diversity detected in a Yellowstone National Park hot spring environment [J]. Proc Natl Acad Sci,1994,91:1609-1613.
[77]Torsvik V, Goksoyr J, Daae FL. High diversity in DNA of soil bacteria [J]. Appli Environ Microbiol,1990,56:782-787.
[78]Amans RI, Luding E, Schleifer KH. Phylogenetic identifiction and in situ detection of individual microbial cells without cultivation [J]. Microbial,1995, 66:4547-4554.
[79]Pace NR. A molecular viewer of microbial diversity and the biosphere [J]. Science,1997,276:734-740.
[80]Bach HJ, Errampalli D, Leung KT, et al. Specific detection of the gene for the extracellular neutral protease of Bacillus cereus by PCR and blot hybridization [J]. Appl Environ Microbiol,1999,65:3226-3228.
[81]Suzuki MT, Rappe MS, Haimberger ZW, et al. Bacterial diversity among small-subunit rRNA gene clones and cellular isolates from the same seawaters ample [J]. Applied and Environmental Microbiology,1997,63(3):983-989.
[82]Dai X, Zhou H, Cai CH, et al. A preliminary study on the novel taxa of bacteria in marine sediments [J]. Acta Scientiarum Naturalium Universitatis Suntvatseni, 2001,40(6):51-54.
[83]Dai X, Zhou H, Cai CH, et al. A preliminary study on the diversity of bacteria in the South China Sea Sedimentts as determined by 16S rRNA gene analysis [J]. Progress of Natural Science,2002,12(5):479-484.
[84]Jutta K, Oliver P, Martin HS, et al. Sulfate-reducing bacterial community response to carbon source amendments in contaminated aquifer microcosms [J]. FEMSMicrobiologv Ecologv,2002,42:109-118.
[85]Rasmussen LD, Ekelund F, Hansen LH. Group-specific PCR primers to amplify 24s a-subunit rRNA genes from Kinetoplastida (Protozoa) used in denaturing gradient gel electrophoresis [J]. Microb Ecol,2001,42:109-115.
[86]Beja 0, Sazuki MT, Heideberg JE, et al. Unsuspected diversity among marine aerobic anoxygenic phototrophs [J]. Nature,2002,415(6287):630-633.
[87]杨官品,茅云翔.环境细菌宏基因组研究及海洋细菌生物活性物质BAC文库筛选[J].青岛海洋大学学报,2001，31：718-722.
[88]Rondon MR, August PR, Bettermann AD, et al. Cloning the soil metagenome:a strategy for accessing the genetic and functional diversity of uncultured microorganisms [J]. Appl Environ Microbiol,2000,66:2541-2547.
[89]周世水,丁金国,姚汝华.海洋微生物药物的开发和应用[J].工业微生物,2002,2
[90]李渤,王乃力,姚新生,胡江春,王书锦.海洋微生物中具有引起稻瘟霉孢子形态变异活性的菌株筛选.沈阳药科大学学报,2002，4
[91]刘志鸿,程力,牟海津.海洋微生物活性物质的研究概况.中国水产科学,1999,4
[92]孙谧,洪义国,李勃生,修朝阳.海洋微生物低温酶特性及其在工业中的潜在用途.海洋水产研究,2002，3
[93]安鑫龙,周启星,刑光敏.海洋微生物在海洋污染治理中的应用现状.水产科学,2006,2
[94]Deming JW. Deep ocean environmental biotechnology [J]. Curr Opin Biotechnol, 1998,9:283-287.
[95]姜英,王秉瑞.抗生素耐药性机理研究进展[J].微生物学免疫学进展,2003,31(3)：74-76.
[96]Knight V, Sanglier JJ, Ditullio D, et al. Diversifying microbial natural products for drug discovery [J]. Appl Microbiol Biotechnol,2003,62(5-6):1-28.
[97]Simidu U, Lee WJ, Kogure K. Comparison of different techniques for determing plate counts of marine bacteria [J]. Bull Jpn Soc Sci Fish,1983,49:1199-1203.
[98]Richard EH, James AZ, Jeffrey DG, et al. Rapid method to estimate the presence of secondary metabolites in microbial extracts [J]. Appl Environ Microbial, 2001, 67(1):371-376.
[99]Amann R, Ludwig W. Ribosomal RNA-targeted nucleic acid probes for studies in microbial ecology [J]. FEMS Microbiol Rev,2000,24(5):555-565.
[100]林敏,宁喜斌.具抑菌活性海洋微生物的筛选[J].微生物学杂志,2005,(5)：23-25.
[101]Acar JF. The disc susceptibility test [Z]. In:Lorian V(ed) Antibiotics in laboratory medicine,1980.
[102]罗文新,陈晓佳,黄耀坚,等.来自海洋的链霉菌抑菌活性与其培养条件的关系[J].微生物学通报,1998,25(6)：325-332.
[103]Lee A, Langer R. Shark cartilage contains inhibitors of tumor angiogenesis [J]. Science,1983,221(4616):1185-1187.
[104]陈建鹤,焦炳华,缪为民,等.中国姥鲨软骨源新生血管抑制因子(Sp8)的分离纯化及生物学活性[J].第二军医大学学报,2000，21(2)：107-110.
[105]Damiani G, Amedeo P, Bandi C, et al. Bacteria indentification by PCR based techniques [M]. Microbial Genome Methods, CRC Press, 1996.
[106]O'Sullivan LA. Fuller KE. Thomas EM, et al. Distribution and culturability of the uncultivated "AGG58 cluster" of the Bacteriodetes phylum in aquatic environments [J]. FEMS Microbiol Ecol,2004, 47:359-370.
[107]Fry NK, Warwick S, Saunders NA, et al. J Gen Microbiol, 1991, 137:1215-1222.
[108]Devereux R, He SH, Doyle CL, et al. J Bacteriol,1990,172:3609-3619.
[109]Woese CR. Bacterial evolution [J]. Microbiol Rev,1987,51:221-271.
[110]Pace NR, Stahl DA, Lane DL, et al. The analysis of natural microbial populations by rRNA suquences [J]. Adv Microbiol Ecol,1986,9:1-55.
[111]vov Dohren H, Dieckmann R, et al. The nonribosomal code [J]. Chem Biol, 1999,6:273-279.
[112]明镇寰,潘建伟,朱睦元.非核糖体多肽合成酶进展[J].生物化学与生物物理进展,2002,29(5)：667-669.
[113]Stachelhaus T, Mooty HD, Marahiel MA. The specificity-conferring code of adenylation domains in nonribosomal peptide synthetases [J]. Chem Biol,1999, 6:493-505.
[114]Cane DE, Walsh CT. The parallel and convergent universes of polyketide synthases and nonribosomal peptide synthetases [J]. Chem Biol,1999, 6:319-325.
[115]Lambalot RH, Walsh CT. A new enzyme superfamily-the phosphopantetheinyl transferases [J]. Chem Biol,1996,3:923-936.
[116]Grunewald J, Marahiel MA. Chemoenzymatic and template-directed synthesis of bioactive macrocyclic peptides [J]. Microbiol Mol Biol Rev,2006,70(1):121-146.
[117]Stachelhaus T, Mooty HD, Bergendahl V, et al. Pepetide bond formation in nonribosomal peptide biosynthesis [J]. J Biol Chem,1998,272:22773-22781.
[118]Mootz HD, Schwarzer D, Marahiel MA. Ways of assembling complex natural products on modular nonribosomal peptide synthetases [J]. Chem Bio Chem, 2002,3(6):490-504.
[119]Becker JE, Moore R, Moore BS. Cloning, sequencing, and biochemical characterization of the nostocyclopeptide biosynthetic gene cluster.molecular basis for imine macrocyclization [J]. Gene,2004,325:35-42.
[120]Cramer RA Jr, Gamcsik MP, Brooking RM, et al. Disruption of a nonribosomal peptide synthetase in Aspergillus fumigatus eliminates gliotoxin production [J]. Eukaryot Cell,2006,5(6):972-801.
[121]Sharma A, Johri BN. Growth promotinginfluence of siderophore-producing Pseudomonas strains GRP3A and PRS9 in maize (Zea mays L.) under iron limiting conditions [J]. Microbiol Res,2003,158:243-248.
[122]Picosi S, Valladares A, Flores E, et al. Nitrogen-reulated genes for the metabolism of cyanophycin, a bacterial nitrogen reserve polymer [J]. J Bio Chem,2004,279(12):11582-11592.
[123]Liu F, Garneau S, Walsh CT. Hybrid nonribosomal peptide-polyketide interfaces in epothilone biosynthesis:minimal requirements at N and C termini of EpoB for elongation [J]. Chem Bio,2004,11:1533-1542.
[124]Miao V, Coeffet-Le Gal MF, Brian P.. et al. Daptomycin biosynthesis in Streptomyces roseosporus.cloning and analysis of the gene cluster and revision of peptide stereochemistry [J]. Microbiology,2005,151:1507-1523.
[125]Schneider A, Stachelhaus T, Marahiel MA. Targeted alteration of the substrate specificity of peptide synthetases by rationalmodule swapping [J]. Mol Gen Genet,1998,257:308-318.
[126]diDuilio M. The origin of peptide synthesis:On some molecular fossils identified through comparison of protein sequences [J]. Biosystems,1996,
39:169.
[127]Xu HS, Roberts N, Singleton FL, et al. Survival and viability of nonculturable Escherichia coli and Vibrio cholerae in the estuarine and marine environment [J]. Microbial Ecology,1982,8:313-323.