深海放线菌Streptomyces sp.WP6次生代谢产物的研究

目的 研究深海来源链霉菌Streptomyces sp.WP6具有生物活性的次级代谢产物.方法 采用正相硅胶柱色谱、反相硅胶柱色谱、Sephadex LH-20凝胶柱色谱、TLC薄层制备层析色谱、中压快速制备层析色谱和高压液相色谱等方法分离纯化次生代谢产物,采用13C NMR、1H NMR、HMBC、HSQC、1H-1H COSY、LC-MS等波谱技术鉴定结构.结果 从该菌株发酵产物中分离得到8个次级代谢产物,分别鉴定为:邻苯二甲酸二(2-乙基己基)酯(1)、N-乙酰基酪胺(2)、脯氨酸-亮氨酸(3)、N-乙酰色胺(4)、吲哚-3-甲酸(5)、苯乙酸(6)、环(脯氨酸-苯丙氨酸)(7)、(4S)-4,10二羟基-10甲基-2烯-1,4内酯(8).结论 首次从Streptomyces sp.WP6发酵产物中分离到上述8个次生代谢产物.     

中国南海黑乳海参共附生真菌Phialemonium sp.次生代谢产物的鉴定

［］　目的　对中国南海黑乳海参的共附生单胞瓶霉属真菌(Phialemonium sp.)的次生代谢产物进行研究。 方法　运用硅胶柱色谱、Sephadex LH-20凝胶柱色谱、高效液相色谱等分离手段对真菌Phialemonium sp.的乙酸乙酯提取物进行分离纯化,利用现代波谱技术结合文献报道进行结构鉴定。 结果　共分离得到2个缩酚酸环醚化合物,emeguisin A和aspergillusidone C。结论　本研究是对黑乳海参共附生真菌Phialemonium sp.次生代谢产物的首次报道,2个缩酚酸环醚成分均为首次从该种真菌中分离得到。     

深海细菌Psychrobacter submarinus 1A01998的次级代谢产物研究

目的 对来自深海沉积物中的海洋细菌Psychrobacter submarinus1 A01998的次级代谢产物进行研究.方法 采用大孑L吸附树脂、硅胶、ODS和Sephadex LH-20凝胶等柱层析色谱分离方法,对海洋细菌Psychrobacter submarinus1 A01998进行次级代谢产物的分离纯化,根据1H NMR、13C NMR和MS等波谱数据分析并结合相关文献对化学结构进行鉴定.结果 从深海细菌Psychrobacter submarinus 1A01998的发酵粗提物中分离得到4个化合物,分别鉴定为环(L-脯氨酸-L-酪氨酸)(1)、环(D-脯氨酸-L-酪氨酸)(2)、3-吲哚甲醛(3)、2'-O-甲氧基尿嘧啶核苷(4).结论 化合物1～4均为首次从该属中分离得到,其中化合物1、2为环二肽类化合物.     

深海真菌Penicillium sp.Z13次级代谢产物的研究

目的：研究海洋真菌Penicillium sp.Z13次级代谢产物。方法：采用凝胶柱,ods柱,反相高效液相制备色谱法等,对该真菌发酵产物的乙酸乙酯提取物进行分离纯化,采用核磁共振氢谱和碳谱等方法,确定化合物的结构。结果：从海洋真菌Penicillium sp. Z13的乙酸乙酯提取物中分离得到10个化合物,分别鉴定为6-desmethylmonacolin-J-15(1), 3,5-dihydroxy-7-(8-hydroxy-2-met-hy-hexahydronaph-thalen-1-yl)heptanoic acid(2)和penicopeptide A(3),圆弧菌素(4)和圆弧菌醇(5),viridicatol (6)和viridicatin (7),吲唑(8),吲哚乙酸(9),环(异亮氨酸-脯氨酸)(10)。结论：海洋真菌Penicillium sp. Z13可以产生结构多样的次级代谢产物。     

新的低聚肽降压药

从无花果得到的低聚肽L-丙氨酸-L-缬氨酸-L-天冬酰胺-L-脯氨酸-L-异亮氨酸-L-精氨酸,L-亮氨酸-L-酪氨酸-L-脯氨酸-L-缬氨酸-L-赖氨酸,L-亮氨酸-L-缬氨酸-L-精氨酸及其加成盐都是新化合物。磨碎无花果肉,分离固体物质。无花果汁或含从果汁分离得到(分子量达10000)的部位的溶液,或二者的浓缩物是含上述有效成分的降压药。此种从无花果得到的低聚肽为极好的降压药。口服或非肠道内(如腹腔注射)给药。剂     

海洋放线菌Nocardiopsis sp.SCSIO 11492的抗肿瘤活性成分研究

目的 研究海洋放线菌Nocardiopsis sp.SCSIO 11492的抗肿瘤活性成分.方法 采用ODS反相硅胶中压柱色谱、正相硅胶柱色谱、Sephadex LH-20凝胶柱色谱等方法分离纯化次级代谢产物;13 C NMR、1 H NMR、HSQC、1 H-1 HCOSY、HMBC等波谱技术并结合电喷雾电离质谱法鉴定结构;MTT法进行体外抗肿瘤活性测试.结果 从海洋放线菌Nocardiopsis sp.SCSIO 11492的乙酸乙酯萃取物中分离得到8个化合物,分别为2′-脱氧腺苷(1)、2′-脱氧胸腺嘧啶核苷(2)、2′-脱氧尿苷(3)、尿嘧啶核苷(4)、1-棕榈酰-3-β-D-半乳糖甘油酯(5)、环(亮-缬)二肽(6)、环(缬-脯)二肽(7)、环(脯-脯)二肽(8).其中化合物5对小鼠单核巨噬细胞白血病细胞RAW264.7显示出较好的生长抑制活性(IC50=10.9μmol/L),其他化合物均没有表现出明显的细胞毒活性(IC50>20.0μmol/L).结论 以上8个化合物均为首次从该菌中分离得到,其中化合物5可能是该菌的抗肿瘤活性成分.     

海洋细菌Halomonas elongate的次级代谢产物研究

目的 对具有抗肿瘤活性的海洋细菌Halomonas elongate进行次级代谢产物的分离、纯化及抗肿瘤活性研究.方法 通过大孔吸附树脂、硅胶、ODS、Sephadex LH-20凝胶柱色谱等分离纯化手段,对H.elongate的次级代谢产物进行分离纯化;根据1H NMR谱、13C NMR谱及MS分析,并结合文献确定结构:采用MTT法对分离得到的次级代谢产物进行活性检测.结果 从海洋细菌H.elongate的发酵粗提物中分离鉴定3个化合物,分别为胸腺嘧啶-2 '-脱氧核苷(1)、环(L-甘氨酸-L-脯氨酸)二肽(2)、1-(2'-脱氧-β-D-赤式-呋喃戊糖)-1氢-1,2,4-三嗪酮(3),活性检测结果显示3个化合物对人肺癌细胞H1299生长活性均无显著抑制作用(IC50＞20 μμmol/L).结论 化合物1～3均为首次从Halomonas中分离得到.     

中国南海黑乳海参共附生白色侧齿霉菌中的甾体类成分

目的对中国南海黑乳海参共附生白色侧齿霉菌Engyodontium album的次生代谢产物进行研究。方法运用硅胶柱色谱、Sephadex LH-20凝胶柱色谱、高效液相色谱等分离手段对白色侧齿霉菌Engyodontium album的乙酸乙酯提取物进行分离纯化,根据现代波谱技术结合文献报道进行结构鉴定。结果共分离得到5个化合物,分别鉴定为:(22E,24R)-5α,8α-过氧化麦角甾-6,22-二烯-3β-醇(1)、(22E,24R)-麦角甾-5,7,22-三烯-3β-醇(2)、(22E,24R)-麦角甾-7,22-二烯-3β,5α,6β-三醇(3)、3β,5α-二羟基-(22E,24R)-麦角甾-7,22-二烯-6-酮(4)、3β,5α-二羟基-6β-甲氧基-麦角甾-7,22-二烯(5)。结论本研究是对黑乳海参共附生白色侧齿霉菌次生代谢产物的首次报道,这5个化合物均为首次从白色侧齿霉菌Engyodontium album中分离得到。     

中国广西涠洲岛海域海绵Callyspongia sp.的化学成分研究

目的:对采自中国广西涠洲岛海域海绵Callyspongia sp.的化学成分进行分离和结构鉴定。方法:采用硅胶薄层色谱、柱色谱、凝胶渗透色谱LH-20,半制备HPLC等方法对采自中国广西涠洲岛海域海绵Callyspongia sp.的次生代谢产物进行化学分离,并通过核磁共振谱(1D、2D NMR)、质谱 (ESI - MS)等方法,确定化合物的结构。结果:共分离得到8个化合物,分别为牛磺酸（1）、乙酰牛磺酸（2）、N-甲基吡啶-2-羧酸盐（3）、zooanemonin（4）、N-丁基乙酰胺（5）、尿嘧啶（6）、3-甲基尿嘧啶（7）、苯甲酸（8）。结论:化合物1-5和7-8均为首次从该属海绵中分离得到。     

蜚蠊肠道放线菌WA23-4-4的抗金黄色葡萄球菌次级代谢产物研究

目的 研究蜚蠊肠道内生放线菌WA23-4-4的抗金黄色葡萄球菌次级代谢产物.方法 采用硅胶柱色谱、葡聚糖凝胶柱色谱、半制备高效液相等方法分离纯化菌株WA23-4-4次级代谢产物;采用紫外光谱(UV)、红外光谱(IR)、核磁共振光谱(NMR)、质谱(MS)等技术鉴定结构;微量肉汤稀释法体外测定抗金黄色葡萄球菌活性.结果 ...     

Estimation of boswellic acids in herbal formulations containing Boswellia serrata extract and comprehensive characterization of secondary metabolites using UPLC-Q-Tof-MSe

Boswellia serrata is a widely used herb in Indian systems of medicine and is well known for its potential medicinal properties. A chromatographic method was developed for the analysis and quantification of six boswellic acid marker compounds, i.e., keto boswellic acid (1), 3-O-Acetyl 11-keto β-boswellic acid (2), ɑ-Boswellic acid (3), β-Boswellic acid (4), 3-O-Acetyl-ɑ-boswellic acid (5) and 3-O-Acetyl-β-boswellic acid (6) in commercial herbal products containing B. serrata as an ingredient. Combining UPLC with Q-Tof-MS/MS makes the better identification of secondary metabolites and adulterants in the herbal formulations containing B. serrata in rapid time using fragmentation approach than the traditional approaches. In this study quantification of boswellic acids with UPLC-PDA method was performed as per the pharmacopeia guidelines. Furthermore, minor phytochemical constituents were identified and characterized with the help of LC-Q-Tof-MS/MS fragmentation data and various isoforms of boswellic acids and tirucallic acids in B. serrata oleo-gum-resin extract were identified.     

Production and purification of a bioactive substance against multi-drug resistant human pathogens from the marine-sponge-derived Salinispora sp.

ObjectiveTo isolate, purify, characterize, elucidate structure and evaluate bioactive compounds from the sponge-derived Salinispora sp. FS-0034.MethodsThe symbiotic actinomycete strain FS-0034 with an interesting bioactivity profile was isolated from the Fijian marine sponge Theonella sp. Based on colony morphology and obligatory requirement of seawater for growth, and mycelia morphological characteristics the isolate FS-0034 was identified as a Salinispora sp. The bioactive compound was identified by using various spectral analysis of ultraviolet, high resolution electrospray ionization mass spectroscopy, ¹H nuclear magnetic resonance, correlated spectroscopy and heteronuclear multiple bond coherence spectral data. A minimum inhibitory concentration assay were performed to evaluate the biological properties of the pure compound against multi-drug resistant pathogens.ResultsBioassay guided fractionation of the ethyl acetate extract of the culture of Salinispora sp. FS-0034 by different chromatographic methods yielded the isolation of an antibacterial compound, which was identified as rifamycin W (compound 1). Rifamycin W was reported for its potent antibacterial activity against methicillin-resistant Staphylococcus aureus, wild type Staphylococcus aureus and vancomycin-resistant Enterococcus faecium and displayed minimum inhibitory concentrations of 15.62, 7.80 and 250.00 μg/mL, respectively.ConclusionsThe present study reported the rifamycin W from sponge-associated Salinispora sp. and it exhibited appreciable antibacterial activity against multi-drug resistant human pathogens which indicated that sponge-associated Actinobacteria are significant sources of bioactive metabolites.     

放线菌野生株代谢功能的核糖体工程改造与新产抗肿瘤活性产物研究

目的 利用核糖体工程技术,改造放线菌次级代谢功能,筛选获得抗肿瘤活性突变株,并对突变株新产活性产物进行研究。方法 以海洋来源无活性放线菌野生株HLF-39和HLF-43为出发菌,通过单菌落挑选与平板划线培养,分离纯化链霉素抗性突变株,通过摇床液态发酵和发酵提取物乙酸乙酯萃取样品的抗肿瘤活性筛选,获得抗肿瘤活性突变株;采用活性跟踪与微量预试先导 放大实验制备组合的实验模式,与原始菌样品对照比较,在快速确定差异活性斑点基础上,组合各种色谱技术,分离纯化突变株新产抗肿瘤活性产物;根据理化性质和波谱数据鉴定化合物结构;采用MTT法测评抗肿瘤活性。结果 链霉素对HLF-39和HLF-43的最低抑菌浓度分别为3和10 mg·L^-1。经抗性筛选,得到对链霉素产生抗性的HLF-39突变株28株、HLF-43突变株204株。在所得突变株中,3株HLF-39突变株和14株HLF-43突变株有抗肿瘤活性,100 mg·L^-1样品对K562细胞的抑制率大于20%;其中,4株分别对11、100、200和300 mg·L-1链霉素产生抗性的HLF-43突变株CHS-21101、CHS-210010、CHS-220002和CHS-230001活性显著,100 mg·L^-1样品对K562细胞的抑制率分别达59.5%、50.0%、44.1%和59.6%。从CHS-21101发酵物中分离得到2个该突变株新产抗肿瘤活性产物,并分别鉴定为环(4-羟脯-亮)二肽（1）和phencomycin（2）。化合物1和2对K562细胞呈抑制活性,100 mg·L^-1抑制率分别为33.9%和21.4%。结论 利用核糖体工程抗性筛选技术,从2株无活性放线菌野生菌株成功筛选得到17株具有抗肿瘤活性的链霉素抗性突变株,从其中1株发酵物中分离鉴定了2个该突变株新产抗肿瘤活性产物。用核糖体工程技术改造无活性放线菌野生株的次级代谢功能,可筛选获得活性突变株并提供深入研究新产活性产物,从中筛选新药及其先导结构,从而拓展药源放线菌活性菌株新资源。     

棘体海绵Acanthella cavernosa中次生代谢产物的分离鉴定

目的对中国南海棘体海绵Acanthella cavernosa的次生代谢产物进行分离鉴定。方法运用硅胶柱色谱、凝胶柱色谱、高效液相色谱等分离手段,对棘体海绵Acanthella cavernosa的丙酮提取物进行分离纯化,根据现代波谱技术结合文献报道进行结构鉴定。结果和结论共分离得到7个化合物,分别鉴定为kalihinene X~Z(1~3)、(R)-3,5-diethyl-5-[(S,E)-2-ethylhex-3-en-1-yl]furan-2(5H)-one(4)、methyl(2Z,6R,8S)-4,6-diethyl-3,6-epoxy-8-methyldodeca-2,4-dienoate(5)、methyl(2Z,6R,8R,9E)-3,6-epoxy-4,6,8-triethyl-2,4,9-dodecatrienoate(6)、methyl(2Z,6R,8S)-3,6-epoxy-4,6,8-triethyldodeca-2,4-dienoate(7)。化合物4~7为首次从Acanthella属中分离得到的聚酮类化合物。     

Characterization of cytotoxic compound from marine sediment derived actinomycete Streptomyces avidinii strain SU4

Objective

To investigate the cytotoxic activity of actinomycete isolated from marine sediment.

Methods

In the present study the DNA was isolated and the ITS region of 16s rRNA was amplified by polymerase chain reaction, using two universal bacterial primers, 1492R (5′-GGTTACCTTGTTAC GACTT-3′) and Eubac27F (5′-AGAGTTTGATCCTGGCTC AG-3′). The amplified products were purified using TIANgel mini purification kit, ligated to MD18-T simple vector (TaKaRa), and transformed into competent cells of Escherichia coli DH5α. 16S rRNA gene fragment was sequenced using forward primer M13F (-47) and reverse primer M13R (-48). Blast search sequence similarity was found against the existing non-redundant nucleotide sequence database thus, identified as Streptomyces sp SU, Streptomyces rubralavandulae strain SU1, Streptomyces cacaoi strain SU2, Streptomyces cavourensis strain SU3, Streptomyces avidinii strain SU4, Streptomyces globisporus strain SU5, Streptomyces variabilis strain SU6, Streptomyces coelicolor strain SU 7. Among the eight identified isolates, one actinomycete Streptomyces avidinii strain SU4 was selected for further study.

Results

Crude extract of the actinomycete isolate exhibited IC₅₀ in 64.5 µg against Hep-2 cell line, 250 µg in VERO cell line. This value is very close to the criteria of cytotoxicity activity for the crude extracts, as established by the American National Cancer Institute (NCI) is in IC₅₀ < 30 µg/mL. The GC MS analysis showed that the active principle might be 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester (12.17%), isooctyl phthalate (15.29%) with the retention time 15.642 and 21.612, respectively.

Conclusions

This study clearly proves that the marine sediment derived actinomycetes with bioactive metabolites can be expected to provide high quality biological material for high throughout biochemical and anticancer screening programs. These results help us to conclude that the potential of using metabolic engineering and post genomic approaches to isolate more bioactive compounds and make their possible commercial application is not far off.     

水母雪莲内生真菌Aspergillus sp. TPXq 的次级代谢产物研究

目的 研究藏药水母雪莲内生真菌Aspergillus sp. TPXq的次级代谢产物。方法 菌种采用震荡培养箱扩大培养,经反复硅胶柱色谱、Sephadex LH-20凝胶柱色谱、开放ODS柱色谱、高效液相色谱等方法进行分离纯化;并根据化合物的理化性质及波谱数据鉴定结构。结果 从来自藏药水母雪莲的一株内生真菌Aspergillus sp. TPXq的培养液中分离得到8个环二肽和1个生物碱类化合物。分别鉴定为3-异丁基-吡咯并哌嗪-2,5-二酮（Ⅰ）、3-异戊基-吡咯并哌嗪-2,5-二酮（Ⅱ）、3-仲丁基-吡咯并哌嗪-2,5-二酮（Ⅲ）、3-苄基-吡咯并哌嗪-2,5-二酮（Ⅳ）、3-苄基-6-对羟基苄基-2,5二酮（Ⅴ）、3,6-二甲基哌嗪-2,5-二酮（Ⅵ）、3-异丁基-6-异丙基哌嗪-2,5-二酮（Ⅶ）、3-异丁基-6-苄基哌嗪-2,5-二酮（Ⅷ）、Chaetominine（Ⅸ）。结论 其中化合物Ⅳ~Ⅷ为曲霉属Aspergillus.sp真菌中首次分离得到。细胞毒测试结果表明,以上化合物Ⅰ~Ⅷ对于A549和MCF-7肿瘤细胞均显示弱细胞毒活性（>50μg/mL）,化合物Ⅸ对A549和MCF-7肿瘤细胞均具有显著的细胞毒活性,IC₅₀值分别为0.18μg/mL和0.89μg/mL。      

Antimicrobial secondary metabolites from marine gastropod egg capsules and egg masses

Marine organisms have attracted special attention in the last three decades for their ability to produce interesting pharmacological active compounds. Even though all marine organisms have the potential to produce antimicrobial secondary metabolites, the gastropod has the vital sources of secondary metabolites particularly their egg capsule which has the promising antimicrobial secondary metabolites. In the present review, we intend to focus on marine secondary metabolites from marine gastropod egg capsule. The following compounds i.e. Kabiramid C, Aplysianin E, Aplysianin A, Thisaplysianin E and Tyrian purple have been documented in egg capsule of various gastropod and most of the antimicrobial secondary metabolites have not been isolated from the egg capsule because of the odious, and complex chemical structure. Stability of the compounds is unknown.