Effects of the piezo-tolerance of cultured deep-sea eel cells on survival rates, cell proliferation, and cytoskeletal structures

We investigated the pressure tolerance of deep-sea eel (Simenchelys parasiticus; habitat depth, 366–2,630 m) cells, conger eel (Conger myriaster) cells, and mouse 3T3-L1 cells. Although there were no living mouse 3T3-L1 and conger eel cells after 130 MPa (0.1 MPa = 1 bar) hydrostatic pressurization for 20 min, all deep-sea eel cells remained alive after being subjected to pressures up to 150 MPa for 20 min. Pressurization at 40 MPa for 20 min induced disruption of actin and tubulin filaments with profound cell-shape changes in the mouse and conger eel cells. In the deep-sea eel cells, microtubules and some actin filaments were disrupted after being subjected to hydrostatic pressure of 100 MPa and greater for 20 min. Conger eel cells were sensitive to pressure and did not grow at 10 MPa. Mouse 3T3-L1 cells grew faster under pressure of 5 MPa than at atmospheric pressure and stopped growing at 18 MPa. Deep-sea eel cells were capable of growth in pressures up to 25 MPa and stopped growing at 30 MPa. Deep-sea eel cells required 4 h at 20 MPa to finish the M phase, which was approximately fourfold the time required under atmospheric conditions.     

太平洋帕里西维拉海盆细菌多样性的非培养的初步分析

从环境中直接提取总DNA ,构建了含 32个有效转化子的太平洋帕里西维拉 (PareceVela)海盆 5 0 10米深处底泥的细菌 16SrRNA基因文库。测序结果表明 ,可以将 32条有效序列分为 17个不同的分类单元。大部分序列与已知细菌类群的 16SrDNA序列相似性较高 ,归属于Proteobacteria的gamma亚群、alpha亚群和海洋非培养细菌类群 ,主要分布在Pseudoalteromonas属、Halomonas属、Alcanivorax属、Photobacterium属、Acinetobacter属 ;部分序列与已知细菌类群的 16SrDNA序列同源性较低 ,可能代表新的分类单位。研究结果表明 ,帕里西维拉海盆不仅含有丰富的微生物物种 ,并且存在尚未被认识的新物种     

深海微生物的研究进展

从生态学的角度介绍了深海微生物的营养来源、生物多样性及相关研究方法并展望深海微生物资源的开发前景。     

新颖深海微生物酯酶EstC11的酶学性质及其在手性拆分乙酸苏合香酯中的应用

【背景】手性乙酸苏合香酯是重要的手性香料产品,在食品及精细化工等领域都有重要的应用。酶催化不对称合成手性乙酸苏合香酯产品具有极好的工业应用前景。【目的】研究酯酶EstC11的基本酶学性质及其在制备手性乙酸苏合香酯中的应用。【方法】对来自西太平洋深海热液口芽孢杆菌Bacillus sp.CX01中的新颖微生物酯酶基因EstC11进行克隆、表达及酶学性质鉴定。通过对p H、温度、有机溶剂等反应条件的优化提高酯酶手性拆分乙酸苏合香酯的光学纯度。【结果】酯酶EstC11的最适反应p H为8.5,最适温度为25°C,一些金属离子和有机溶剂对酯酶EstC11的水解活性具有不同程度的抑制作用。通过对反应条件的优化,在最适反应条件下(p H 9.0 50 mmol/L Tris-HCl,20°C,50 mmol/L底物浓度)反应3 h后,(R)-乙酸苏合香酯的光学纯度达98%,得率为39%。【结论】通过对酯酶拆分条件的优化,手性拆分乙酸苏合香酯生成(R)-乙酸苏合香酯的光学纯度明显提高,为酯酶EstC11在工业化上的应用奠定了基础。     

Nitrile hydrolysing activities of deep-sea and terrestrial mycolate actinomycetes

Nitrile metabolising actinomycetes previously recovered from deep-sea sediments and terrestrial soils were investigated for their nitrile transforming properties. Metabolic profiling and activity assays confirmed that all strains catalysed the hydrolysis of nitriles by a nitrile hydratase/amidase system. Acetonitrile and benzonitrile, when used as growth substrates for enzyme induction experiments, had a significant influence on the biotransformation activities towards various nitriles and amides. The specific activities of selected deep-sea and terrestrial acetonitrile-grown bacteria against a suite of nitriles and amides were higher than those of the only other reported marine nitrile-hydrolysing R. erythropolis, isolated from a shallow sediment. The increase of nitrile chain length appeared to have negative influence on the nitrile hydratase activity of acetonitrile-grown bacteria, but the same was not true for benzonitrile-grown bacteria. The nitrile hydratases and amidases were constitutive in 10 of the 16 deep-sea and terrestrial actinomycetes studied, and one strain showed an inducible hydratase and a constitutive amidase. Most of the deep-sea strains had constitutive activities and showed some of the highest activities and broadest substrate specificities of organisms included in this study. This revised version was published online in August 2006 with corrections to the Cover Date.     

Sequential changes in cell morphology of an obligately barophilic deep-sea bacterium during its growth at high pressures

Abstract An obligately barophilic bacterium isolated from intestinal content of a deep-sea fish ( Coryphaenoides yaquinae ), which was retrieved from a depth of 6100 m in the Northwest Pacific Ocean, swelled in the early-exponential phase, elongated in the midexponential phase, thinned in the late-exponential phase, and shortened in the stationary phase when incubated both at 82.7 MPa and at 41.4 MPa around the optimum growth pressure. The same changes were observed at any pressures where the strain could grow. We propose to call such morphological changes the SETS (swell-elongate-thin-shorten) phenomenon. These results suggest the possibility that the SETS phenomenon might occur in the growth of the strain at high-pressure deep-sea environments.     

南海深海沉积物中52株真菌的初步分离鉴定及其代谢产物活性

【背景】深海环境复杂多样,蕴含着丰富的真菌资源,为深海真菌结构新颖代谢产物化学及生物活性研究提供了新契机。【目的】对南海深海环境来源真菌进行初步鉴定和代谢产物活性初步研究,旨在筛选和发现潜在的药源真菌新资源,为南海深海真菌资源的进一步开发利用奠定基础。【方法】基于内转录间隔区(ITS)序列测定对分离得到的52株真菌进行初步鉴定,利用纸片扩散法、Solis改良法和DPPH自由基清除法对真菌粗浸膏进行了抗菌(ABA)、卤虫致死(BSL)和抗氧化活性(ABTS)筛选。【结果】52株真菌分布在16个属,其中枝孢菌属(Cladosporium)、曲霉属(Aspergillus)为优势菌群,分别占菌株总数的25.00%、23.08%;32株真菌发酵产物具有抑制至少1种指示菌的活性,其中8株对所有指示菌均有抑制作用;23株具有一定强度的卤虫致死活性,占总数的44.23%,其中有2株活性较显著,IC50值为68.59μg/mL和78.83μg/mL;30株具有DPPH自由基清除活性,占总数的57.69%,其中有9株活性较显著,EC50值低于100μg/mL。【结论】初步认知了南海部分海域深海沉积环境真菌分布和代谢产物活性特征,发现了一批潜在的活性真菌资源,为后续深海真菌代谢产物化学多样性及其功能研究提供支撑。