从烟台附近海域通过挂板取样获得了污损微生物膜样品。利用高通量测序鉴定其中细菌有假交替单胞菌(Pseudoalteromonas sp.),芽孢杆菌(Bacillus sp.),盐单胞菌(Halomonas sp.),嗜冷杆菌(Psychrobacter sp.),肠杆菌(Enterobacter sp.)。通过对微生物膜污损细菌进行发酵培养,结合天然产物分离纯化和核磁共振鉴定技术,共从发酵液中分离并鉴定了9种次级代谢产物,其中4种是二酮哌嗪(Diketopiperazine,DKP)类信号分子,环(脯氨酸-甘氨酸)(1),环(脯氨酸-丙氨酸)(2),环(4-羟基-脯氨酸-亮氨酸)(3),环(脯氨酸-酪氨酸)(4),以及尿嘧啶(5),胸腺嘧啶(6),对羟基苯乙醇(7),十五烷酸(8),邻苯二甲酸二(2-乙基己)酯(9)。研究结果表明,烟台海域污损微生物膜中细菌组成多样,以假交替单胞菌和芽孢杆菌为主;污损细菌产生了多种代谢物,其中包括4种DKP类信号分子,该类物质可能对生物污损的发生具有调控作用,为探究生物污损和生物膜的作用机制奠定了基础。
To investigate the biofouling biofilms, exposure tests were performed in Yantai coastal zone. High throughput sequencing revealed that there were many bacterial species in the biofouling biofilms including Pseudoalteromonas spp., Bacillus spp., Halomonas spp., Psychrobacter spp. and Enterobacter spp. Metabolites chemical purification from the fermentation broth of the bacterial strains was conducted using silica gel column chromatography. Structures of the isolated chemicals were elucidated by NMR. Totally nine chemicals were purified, including four Diketopiperazines: cyclo (Pro-Gly) (1), cyclo (Pro-Ala) (2), cyclo (L-Leu-trans-4- hydroxy-L-Pro) (3) and cyclo (Pro-Tyr) (4). Others were as following: uracil (5), thymine (6), hydroxyphenethyl alcohol (7), pentadecanoic acid (8) and phthalate esters (2-ethyl caproic) (9). This investigation revealed that the bacterial community in biofouling biofilms in Yantai coastal zone was complicated and the major species were Pseudoalteromonas spp. and Bacillus spp. Several metabolites were testified, including four DKPs signal molecules, which might affect the formation ofbiofouling biofilms. This serves as fundamental research in antifouling studies.