海洋抑菌放线菌Y15的筛选、鉴定及其代谢产物理化性质

【目的】筛选产广谱、高效抑菌活性物质的海洋放线菌,为新型抗生素的开发奠定基础。【方法】采用琼脂块法初筛,打孔扩散法复筛,以金黄色葡萄球菌、大肠埃希氏菌、枯草芽孢杆菌和单核细胞增生李斯特菌为指示菌从海泥样品中筛选目标菌株;利用20种指示菌株考查Y10、Y11、Y15、Y16和Y21的抑菌谱;通过形态观察和16S r RNA基因序列分析确定菌株分类地位;以单核细胞增生李斯特菌的OD600值为指标探讨Y15抑菌活性物质对该菌的作用方式;以抑菌活性为指标研究Y15抑菌活性物质的理化性质。【结果】共分离12株具抑菌活性的海洋放线菌,其中Y15抑菌谱最广,对20种指示菌株中的18种具有抑菌活性,并且在4种培养基中均能产生抑菌活性物质。16S r RNA基因序列分析表明Y15属于小单孢菌属,并与Micromonospora endolithica亲缘关系最近。Y15抑菌活性物质在144 h达到最高值为480 AU/m L,在168-216 h保持平衡为320 AU/m L。Y15抑菌活性物质对单核细胞李斯特菌作用方式为杀菌。Y15抑菌活性物质在-20-60°C抑菌活性保持稳定,在80-120°C活性逐渐下降,但在120°C处理30 min仍保留37.5%的活性;在p H 7.0-10.0抑菌活性稳定,在p H 2.0-6.0和p H 11.0-12.0抑菌活性均有所损失;Y15抑菌活性物质对紫外和4种酶(蛋白酶K、胰蛋白酶、木瓜蛋白酶和α-淀粉酶)处理均保持稳定,表明活性物质为非蛋白和非多肽类的抑菌物质。【结论】Y15产生的活性物质具有良好的抑菌谱和抑菌活性,且较为稳定,具有较高的应用价值。     

恩拉霉素生产菌株的遗传改造

【目的】提高杀真菌素链霉菌发酵生产恩拉霉素的产量。【方法】利用定点突变技术,对恩拉霉素生产菌株杀真菌素链霉菌F1中影响细胞次级代谢及抗生素合成的核糖体S12蛋白的编码基因rps L进行改造,将第43位的赖氨酸(Lys)分别替换为天冬酰胺(Asn)和精氨酸(Arg),并对改造菌株L-M1(Asn43)和L-M2(Arg43)的生长特性、抗生素合成以及摇瓶发酵性能进行研究。【结果】与野生型菌株相比,改造菌株的生长特性及生理生化特性均发生了明显的改变:产孢周期明显缩短,野生型菌株在MS培养基中,28°C下需要培养5-7 d后才能产生孢子,而在相同条件下,改造菌株3 d后就能产生大量的孢子;恩拉霉素产量相对提高,摇瓶发酵条件下,改造菌株L-M1(Asn43)和L-M2(Arg43)的恩拉霉素产量分别可达到1 334 U/m L和1 456 U/m L,与野生型菌株F1相比分别提高了11.9%和22.1%。【结论】通过遗传改造,恩拉霉素的产量得到了提高,为其他位点的遗传改造提供了可行性。     

滨海深古菌的研究进展

深古菌(Bathyarchaeota),原名MCG(Miscellaneous Crenarchaeotal Group)古菌,是一类至今未被分离培养的古菌,普遍存在于海洋和陆地环境中。深古菌具有极高的种群多样性,目前已发现多达23个亚群。深古菌具有多种生理生化功能,能降解蛋白质、多聚碳水化合物、脂肪酸、芳香族化合物和甲基化合物等有机质,参与甲烷代谢循环,产乙酸,异化还原亚硝酸盐和硫酸盐,很可能是地球碳元素循环的重要驱动力之一。本文简要概述了深古菌的研究发展历史,阐述了深古菌的分子系统发育分析、分布和基于基因组的生理特征研究的最新进展,总结了滨海深古菌的研究现状,并对滨海深古菌研究的发展方向进行了分析和展望。     

Actinomycetes‐mediated biogenic synthesis of metal and metal oxide nanoparticles: progress and challenges

Actinomycetes‐mediated biogenic synthesis of metal nanoparticles and their antimicrobial activities are well documented. Actinomycetes facilitate both intracellular and extracellular metal nanoparticles synthesis and are efficient candidates for the production of polydispersed, stable and ultra‐small size metal nanoparticles. Secondary metabolites and new chemical entities derived from Actinomycetes have not been extensively studied for the synthesis of metal/metal oxide nanoparticles. The present review focuses on biogenic synthesis of metal nanoparticles from Actinomycetes and the scope for exploring Actinomycetes‐derived compounds (enzymes, organics acids and bioactive compounds) as metal and metal oxide reducing agents for the synthesis of desired nanoparticles. This review also focuses on challenges faced in the applications of nanoparticles and the methods to synthesize biogenic metal nanoparticles with desired physiochemical properties such as ultra‐small size, large surface to mass ratio, high reactivity etc. Methods to evade their toxicity and unique interactions with biological systems to improve their chance as an alternative therapeutic agent in medical and pharmaceutical industry are also discussed.     

Rhodobacteraceae on the marine brown alga Fucus spiralis are abundant and show physiological adaptation to an epiphytic lifestyle

Macroalgae harbour specific microbial communities on their surface that have functions related to host health and defence. In this study, the bacterial biofilm of the marine brown alga Fucus spiralis was investigated using 16S rRNA gene amplicon-based analysis and isolation of bacteria. Rhodobacteraceae (Alphaproteobacteria) were the predominant family constituting 23% of the epibacterial community. At the genus level, Sulfitobacter, Loktanella, Octadecabacter and a previously undescribed cluster were most abundant, and together they comprised 89% of the Rhodobacteraceae. Supported by a specific PCR approach, 23 different Rhodobacteraceae-affiliated strains were isolated from the surface of F. spiralis, which belonged to 12 established and three new genera. For seven strains, closely related sequences were detected in the 16S rRNA gene dataset. Growth experiments with substrates known to be produced by Fucus spp. showed that all of them were consumed by at least three strains, and vitamin B₁₂ was produced by 70% of the isolates. Since growth of F. spiralis depends on B₁₂ supplementation, bacteria may provide the alga with this vitamin. Most strains produced siderophores, which can enhance algal growth under iron-deficient conditions. Inhibiting properties against other bacteria were only observed when F. spiralis material was present in the medium. Thus, the physiological properties of the isolates indicated adaption to an epiphytic lifestyle.     

Ensifer shofinae sp. nov., a novel rhizobial species isolated from root nodules of soybean (Glycine max)

Two bacterial strains isolated from root nodules of soybean were characterized phylogenetically as members of a distinct group in the genus Ensifer based on 16S rRNA gene comparisons. They were also verified as a separated group by the concatenated sequence analyses of recA, atpD and glnII (with similarities ≤93.9% to the type strains for defined species), and by the average nucleotide identities (ANI) between the whole genome sequence of the representative strain CCBAU 251167^T and those of the closely related strains in Ensifer glycinis and Ensifer fredii (90.5% and 90.3%, respectively). Phylogeny of symbiotic genes (nodC and nifH) grouped these two strains together with some soybean-nodulating strains of E. fredii, E. glycinis and Ensifer sojae. Nodulation tests indicated that the representative strain CCBAU 251167^T could form root nodules with capability of nitrogen fixing on its host plant and Glycine soja, Cajanus cajan, Vigna unguiculata, Phaseolus vulgaris and Astragalus membranaceus, and it formed ineffective nodules on Leucaena leucocephala. Strain CCBAU 251167^T contained fatty acids 18:1 ω9c, 18:0 iso and 20:0, differing from other related strains. Utilization of l-threonine and d-serine as carbon source, growth at pH 6.0 and intolerance of 1% (w/v) NaCl distinguished strain CCBAU 251167^T from other type strains of the related species. The genome size of CCBAU 251167^T was 6.2 Mbp, comprising 7,581 predicted genes with DNA G+C content of 59.9 mol% and 970 unique genes. Therefore, a novel species, Ensifer shofinae sp. nov., is proposed, with CCBAU 251167^T (=ACCC 19939^T = LMG 29645^T) as type strain.     

钾离子通道与神经胶质瘤关系的研究进展

钾离子通道为组织细胞内分布最广、种类最多的离子通道,在细胞增殖、分化及肿瘤细胞的侵袭转移中起着关键作用。神经胶质瘤是颅内最多发的恶性肿瘤,目前其主要治疗方式为手术加术后放化疗,术后五年生存率较低,寻找其相关发病机制及化疗靶点具有重要意义。目前已有多项研究表明,多种钾离子通道在胶质瘤中呈特异性高表达,且与胶质瘤的增殖、分化有密切关系,一些钾离子通道可作为胶质瘤的诊断和预防因子,有望成为未来胶质瘤化疗的新靶点,研究钾离子通道与神经胶质瘤的关系对胶质瘤的诊断、预防和治疗有重要意义。本文主要对近年来钾离子通道与神经胶质瘤关系研究的新进展进行综述。     

阿尔茨海默病的发病机制及药物治疗的研究进展

阿尔茨海默病(Alzheimer's disease,AD)是一种慢性进行性神经变性疾病,早期临床表现为近期记忆力下降,后来逐渐发展为生活不能自理最终死亡。目前,有许多种假说阐述其发病机制,如β-淀粉样蛋白、载脂蛋白E、一些金属元素、炎症因子等,但都不全面,也有人认为其可能与朊蛋白有关。部分药物被用于治疗AD,如美金刚、胆碱酯酶抑制剂等,但疗效均不显著。一些新的治疗理念和方法也逐渐被发现,如脑源性神经生长因子(brain-derived neurotrophic factor,BDNF)以及免疫疗法等,但其远期疗效还需进一步证实,且一些副作用还并未被人们所知晓。本文主要对阿尔茨海默病的发病机制及药物治疗的研究进展进行了综述。     

Chemistry of Two Distinct Aeolid Spurilla Species: Ecological Implications

The lipophilic extracts of two marine aeolid nudibranch molluscs of the genus Spurilla collected in distinct geographical areas have been chemically analyzed. The Et₂O extracts of the nudibranchs were dominated by the presence of usual fatty acids and sterols and contained terpenoid compounds 1  –  3 as minor metabolites. Spurillin A ( 1 ) and spurillin B ( 3 ) were new molecules whereas cis‐γ‐monocyclofarnesol ( 2 ) was already reported in the literature as a synthesis product. Interestingly, bursatellin ( 4 ), previously isolated from anaspidean molluscs of the genus Bursatella, was found in the butanol extract of both Spurilla species. Compounds 1  –  4 were not detected in the extracts of the sea‐anemone preys collected together with the molluscs.     

Detection of invasive and cryptic species in marine mussels (Bivalvia,Mytilidae): A chromosomal perspective

Marine mussels illustrate a stunning variability in shape and color. Such variability, added to the scarcity of reliable morphological characters for their identification, can mislead recognition prompting the assignation of specimens of a single species to different ones or incorporate specimens belonging to different taxa into a single one. DNA barcoding is widely used for species identification; however, as this method relies on the previous morphological identification of the specimens, some of the DNA sequences stored in DNA databases are incorrectly assigned to a given species. In view of this uncertainty, further criteria beyond morphological characters and DNA sequences in databases are required to more reliably and accurately identify marine mussels. In this work we mapped ribosomal RNA and histone gene clusters to chromosomes of four species of marine mussels and compared them with those from another eight marine mussel taxa. Specimens of these twelve taxa were also DNA barcoded. Our results clearly demonstrated that the chromosomal analysis of marine mussels could shed light on their identification and, therefore, solve contradictions posed by morphological and molecular data.