低温葡聚糖内切酶产生菌的筛选、鉴定及酶学性质

自黄海长海县附近海泥中分离得到一株产低温葡聚糖内切酶的菌株SWD-28, 经形态学及ITS 序列鉴定为Penicillium cordubense。对该菌株产的粗酶液进行硫酸铵盐析和Sephadex G100柱层析, 比活力达到26.4 U/mg, 提高20.6 倍, 回收率13.1%。得到一个电泳纯的低温葡聚糖内切酶, 分子量为33.1 kD。经圆二色对其结构进行检测, 发现其α-螺旋占49.9%, β-折叠占0.0%, 转角占24.3%, 随机卷曲占25.8%, 呈典型α 螺旋。对其酶学性质进行初步研究, 结果表明其最适pH为5.0, 最适反应温度为35 °C, 在5 °C 酶活力仍能保持60%。     

降解尿酸乳酸菌复合菌系的筛选与菌株组合提升降解效果

【背景】高尿酸症由血液中尿酸含量明显升高而导致,利用乳酸菌对人体的益生作用缓解高尿酸血症越来越受到关注。【目的】获得具有降解尿酸能力的乳酸菌复合菌系与纯培养菌株。【方法】以泡菜为样品来源,以尿酸为底物,采用MRS培养基筛选降解尿酸的乳酸菌复合菌系,通过高效液相色谱法测定复合菌系对尿酸的降解能力。【结果】得到一组乳酸菌复合菌系,当培养温度为37 °C、pH值为6.20、静置培养72 h后复合菌系对尿酸的降解率为12.08%;通过优化培养条件,当该菌系在以牛肉膏为单一氮源、初始pH值为5.00、温度为35 °C的条件下培养72 h,尿酸降解率上升至17.19%,降解率比优化前提高了42.3%;从该菌系中分离出两株具有尿酸降解能力的菌株UA-1与UA-2,它们的尿酸降解率分别为10.85%和8.65%;通过形态学观察和16S rRNA基因序列分析,经鉴定两株菌均为布氏乳杆菌(Lactobacillus buchneri)。将两株单菌组合降解尿酸试验发现,UA-1与UA-2比例为2:1的尿酸降解率为20.2%,比原复合菌系的降解能力提高了67.22%。【结论】研究证明了乳酸菌复合菌系对尿酸的降解能力优于单个菌株,为后续利用乳酸菌复合菌系应用提供了数据支持。     

聚丁二酸丁二醇酯降解菌株的筛选及其降解性能研究

从石油污染土壤中筛选到1株具有聚丁二酸丁二醇酯降解能力的菌株PBS1302,经菌体形态特征、菌落培养特征、生理生化鉴定和16S r DNA基因序列分析,初步鉴定为铜绿假单胞菌(Pseudomonas aeruginosa)。该菌株在培养温度37℃,培养基起始p H 6.8的条件下经6 d的培养,对聚丁二酸丁二醇酯薄膜的降解率可达36.9%。经电子显微镜观察,与降解前相比,聚丁二酸丁二醇酯薄膜表面变得粗糙,出现明显的蚀刻痕迹。     

一株瘤胃纤维素降解菌的分离鉴定及其纤维素降解特性

从蒙古绵羊瘤胃内容物中分离到一株纤维素降解细菌WH-1, 通过形态、生理生化特征、G+C mol%含量和16S rRNA序列分析对分离菌株进行鉴定, 鉴定为溶纤维丁酸弧菌属(Butyrivibrio fibrisolvens)的溶纤维丁酸弧菌(Butyrivibrio fibrisolvens)。同时, 用Mega 4.1软件构建的系统发育树显示分离菌株WH-1与多株溶纤维丁酸弧菌(Butyrivibrio fibrisolvens)的亲缘关系最近。对该菌株纤维素降解特性的初步研究表明：当温度为37°C、     

一株丁草胺降解菌的分离鉴定及其降解特性的研究

利用富集培养技术从长期施用丁草胺的稻田土壤中分离得到能够降解丁草胺的细菌1株, 标记为LYC-1。经形态特征、生理生化特征和16S rRNA序列分析, 将该菌株鉴定为不动杆菌属(Acinetobacter sp.), 菌株LYC-1的最适生长温度为30°C, 最适pH值为7.5。当接种量为5%时, 该菌株在含100 mg/L 的丁草胺无机盐基础培养液中培养7 d后, 可使丁草胺降解达80%以上。     

一株高效苯酚降解菌的选育及降酚性能研究

从一绝缘材料厂的污水中分离得到一株可高效降解苯酚的菌株JY01, 该菌株的形态和理化特征与Bacillus基本相同, 其16S rDNA序列与Bacillus simplex (AM9216370)的相似性为99.01%。在接种量为2%的条件下, 该菌在pH为6.0~9.0和温度为18°C~36°C的范围内保持对苯酚良好的降解能力; 30 h内, 当苯酚浓度为1100 mg/L和1300 mg/L时, 其降解率分别为99.16%和74.76%。这将为进一步采用生物法处理含酚废水提供了可靠的控制条件。     

降解棉秸秆耐热菌株的鉴定及降解条件优化

【目的】鉴定从新疆棉花秸秆高温堆肥中分离出的两株耐热真菌Z1、Z2的属种,并通过优化影响菌株产生纤维素酶的因素来提高菌株对秸秆的降解率。【方法】经形态学和菌株的ITS区克隆与序列分析确定属种,以液体摇瓶发酵产滤纸酶活性(FPA)变化为衡量指标,对Z1、Z2以及二者混合菌(MS)的纤维素酶产生条件进行优化。【结果】菌株Z1为曲霉属烟曲霉(Aspergillus fumigatus Fresen),Z2为蚀丝霉属(Myceliophthora Cost.)。确定Z1以棉秸秆为碳源、以NaNO3为氮源、起始pH 9.5、接种量11%、50°C摇床培养10 d,对棉秸秆降解率为10.19%;Z2以麦秸秆为碳源、以NaNO3为氮源、起始pH 5.5、接种量9%、50°C摇床培养10 d,对麦秆降解率为27.50%;MS以棉花秸秆为碳源、以蛋白胨为氮源、起始pH 5.5、接种量11%、50°C摇床培养10 d,对棉秸秆的降解率为53.45%。【结论】实验表明,MS(Z1、Z2混合)对秸秆的降解效果优于单株菌,降解率达到一半以上,本研究中的两株耐热真菌在降解棉花秸秆、小麦秸秆等农作物废弃秸秆中具有较高的应用价值。     

一株DMP降解菌的分离鉴定及其降解特性

从山东省潍坊市污染河流底泥中筛选到1株能够以酞酸酯(Phthalic acid esters, PAEs)为唯一碳源和能源生长的微生物, 命名为JDC-3, 根据形态学观察、生理生化指标测定和分子生物学鉴定结果, 将该菌株初步鉴定为戴尔福特菌属(Delftia sp.), 以一对简并引物, 首次在该属中扩增出编码邻苯二甲酸双加氧酶的基因片段。同时以邻苯二甲酸二甲酯(Dimethyl phthalate, DMP)为目标测试物, 利用高效液相色谱(HPLC)测定了JDC-3的降解性能, 得出该菌对DMP降解的最佳条件为: pH 7.0~8.0、温度30°C~35°C; 在不同DMP初始浓度下研究了该菌的降解动力学, 结果表明当浓度低于300 mg/L时的降解动力学方程为ln C = - 0.06837 t + A, 半衰期为12.48 h, 当初始浓度不断增加, DMP对JDC-3的抑制能力增强, JDC-3对DMP的降解速率不断下降, 半衰期增大。     

紫外线诱变选育聚丁二酸丁二醇酯（PBS）高降解活性菌株

以PBS降解菌HJ03（Alternariasp．）为出发菌株,通过紫外诱变,透明圈初筛及PBS薄膜复筛,获得一株降解能力增强且对温度和pH耐受力均得到提高的突变株HJ10。与出发菌株相比,HJ10在培养初期气生菌丝少,而培养7d时菌落致密且生长速度较快。经过连续继代培养7代后发现,突变株的降解活力保持了良好的遗传稳定性,其降解率较出发菌株提高百分比达14．4％以上;在最适降解温度范围内（25℃～30℃）和不适宜降解的温度条件下突变菌株Ⅻ10的降解率均高于出发菌株;各pH条件下,突变株对PBS的降解能力明显优于出发菌株,尤其在pH5．0时降解率提高了22．80％。     

三氯乙烯降解菌FT17的分离、鉴定及其降解特性研究

采用水-硅油双相系统, 从辽河流域浑河沈阳段底泥中筛选得到一株三氯乙烯降解菌FT17。综合形态特征、生理生化特征、16S rRNA Blast分析和系统发育分析结果, 将该菌株鉴定为Sporosarcina ginsengisoli。菌株FT17最适生长温度为34°C, 最适生长pH为7.8。苯酚作为共代谢基质可以促进该菌株对三氯乙烯的降解。该菌株的三氯乙烯降解酶在胞内和胞外均存在。采用两种质粒提取方法对该菌株进行质粒检测, 结果均没有发现质粒条带, 推测该菌株的三氯乙烯降解基因位于染色体上。     

Colourful parrot feathers resist bacterial degradation

The brilliant red, orange and yellow colours of parrot feathers are the product of psittacofulvins, which are synthetic pigments known only from parrots. Recent evidence suggests that some pigments in bird feathers function not just as colour generators, but also preserve plumage integrity by increasing the resistance of feather keratin to bacterial degradation. We exposed a variety of colourful parrot feathers to feather-degrading Bacillus licheniformis and found that feathers with red psittacofulvins degraded at about the same rate as those with melanin and more slowly than white feathers, which lack pigments. Blue feathers, in which colour is based on the microstructural arrangement of keratin, air and melanin granules, and green feathers, which combine structural blue with yellow psittacofulvins, degraded at a rate similar to that of red and black feathers. These differences in resistance to bacterial degradation of differently coloured feathers suggest that colour patterns within the Psittaciformes may have evolved to resist bacterial degradation, in addition to their role in communication and camouflage.     

Complement-mediated tail degradation of Neodiplostomum seoulense cercariae

The furcocercus cercariae of Neodiplostomum seoulense (Digenea: Neodiplostomidae) penetrate the skins of tadpoles and shed their tails. The speculated mechanism of this tail loss was physical efforts required to produce a vigorous zigzag motion during skin penetration; no other mechanism has been proposed. We examined the relationship between the host serum and cercarial tail loss. Cercariae of N. seoulense were collected from experimentally infected Segmentina hemisphaerula, and lots of 300 cercariae were cultured in medium 199 contained several types of sera. Cercarial tail degradation was induced in all media, but all the cercariae cultured except those cultured in media containing fetal bovine serum (FBS) died within 48 hr. After 72 hr cultivation in media containing FBS, cercarial tail degradation was induced in 67.0%; in continuous cultivation 13.3% of larvae survived for 7 days. Tail degradation did not occur in the absence of serum and when serum was heat inactivated at 56 degrees Celsius for 30 min. The addition of 20 mM ethylenediaminetetraacetic acid (EDTA) blocked cercarial tail degradation completely. Moreover, the addition of 20 mM MgCl2 restored tail degradation blocked by EDTA. These results suggest that the alternative complement pathway is related with the N. seoulense cercarial tail degradation induced by serum.     

Intracellular localization of proteasomal degradation of a viral antigen.

To better understand proteasomal degradation of nuclear proteins and viral antigens we studied mutated forms of influenza virus nucleoprotein (NP) that misfold and are rapidly degraded by proteasomes. In the presence of proteasome inhibitors, mutated NP (dNP) accumulates in highly insoluble ubiquitinated and nonubiquitinated species in nuclear substructures known as promyelocytic leukemia oncogenic domains (PODs) and the microtubule organizing center (MTOC). Immunofluorescence revealed that dNP recruits proteasomes and a selective assortment of molecular chaperones to both locales, and that a similar (though less dramatic) effect is induced by proteasome inhibitors in the absence of dNP expression. Biochemical evidence is consistent with the idea that dNP is delivered to PODs/MTOC in the absence of proteasome inhibitors. Restoring proteasome activity while blocking protein synthesis results in disappearance of dNP from PODs and the MTOC and the generation of a major histocompatibility complex class I-bound peptide derived from dNP but not NP. These findings demonstrate that PODs and the MTOC serve as sites of proteasomal degradation of misfolded dNP and probably cellular proteins as well, and imply that antigenic peptides are generated at one or both of these sites.     

Habitat degradation impacts black howler monkey (Alouatta pigra) gastrointestinal microbiomes

The gastrointestinal (GI) microbiome contributes significantly to host nutrition and health. However, relationships involving GI microbes, their hosts and host macrohabitats remain to be established. Here, we define clear patterns of variation in the GI microbiomes of six groups of Mexican black howler monkeys (Alouatta pigra) occupying a gradation of habitats including a continuous evergreen rainforest, an evergreen rainforest fragment, a continuous semi-deciduous forest and captivity. High throughput microbial 16S ribosomal RNA gene sequencing indicated that diversity, richness and composition of howler GI microbiomes varied with host habitat in relation to diet. Howlers occupying suboptimal habitats consumed less diverse diets and correspondingly had less diverse gut microbiomes. Quantitative real-time PCR also revealed a reduction in the number of genes related to butyrate production and hydrogen metabolism in the microbiomes of howlers occupying suboptimal habitats, which may impact host health.     

Novel Clostridium populations involved in the anaerobic degradation of Microcystis blooms

Understanding the microbial degradation of Microcystis biomass is crucial for determining the ecological consequences of Microcystis blooms in freshwater lakes. The purpose of this study was to identify bacteria involved in the anaerobic degradation of Microcystis blooms. Microcystis scum was anaerobically incubated for 90 days at three temperatures (15 °C, 25 °C and 35 °C). We used terminal restriction fragment length polymorphism (T-RFLP) analysis of bacterial 16S rRNA genes, followed by cloning and sequencing of selected samples, to reveal the community composition of bacteria and their dynamics during decomposition. Clostridium spp. were found to be the most dominant bacteria in the incubations, accounting for 72% of the sequenced clones. Eight new clusters or subclusters (designated CLOS.1–8) were identified in the Clostridium phylogenetic tree. The bacterial populations displayed distinct successions during Microcystis decomposition. Temperature had a strong effect on the dynamics of the bacterial populations. At 15 °C, the initial dominance of a 207-bp T-RF (Betaproteobacteria) was largely substituted by a 227-bp T-RF (Clostridium, new cluster CLOS.2) at 30 days. In contrast, at 25 °C and 35 °C, we observed an alternating succession of the 227-bp T-RF and a 231-bp T-RF (Clostridium, new cluster CLOS.1) that occurred more than four times; no one species dominated the flora for the entire experiment. Our study shows that novel Clostridium clusters and their diverse consortiums dominate the bacterial communities during anaerobic degradation of Microcystis, suggesting that these microbes'' function in the degradation process.     

ABA modulates the degradation of phosphoenolpyruvate carboxylase kinase in sorghum leaves

Salt stresses strongly enhance the phosphoenolpyruvate carboxylase kinase (PEPC-k) activity of sorghum leaves. This work shows that (1) abscisic acid (ABA) increased the rise in kinase activity in illuminated leaf disks of the non-stressed plant, (2) ABA decreased the disappearance of PEPC-k activity in the dark, (3) two PEPC-k genes expressed in sorghum leaves, PPCK1 and PPCK2, were not up-regulated by the phytohormone and, (4) ABA effects were mimicked by MG132, a powerful inhibitor of the ubiquitin-proteasome pathway. Collectively these data support a role for the ubiquitin-proteasome pathway in the rapid turnover of PEPC-k. The negative control by ABA on this pathway might account for the increase of kinase activity observed in salt-treated plants.     

Predictable bacterial composition and hydrocarbon degradation in Arctic soils following diesel and nutrient disturbance

Increased exploration and exploitation of resources in the Arctic is leading to a higher risk of petroleum contamination. A number of Arctic microorganisms can use petroleum for growth-supporting carbon and energy, but traditional approaches for stimulating these microorganisms (for example, nutrient addition) have varied in effectiveness between sites. Consistent environmental controls on microbial community response to disturbance from petroleum contaminants and nutrient amendments across Arctic soils have not been identified, nor is it known whether specific taxa are universally associated with efficient bioremediation. In this study, we contaminated 18 Arctic soils with diesel and treated subsamples of each with monoammonium phosphate (MAP), which has successfully stimulated degradation in some contaminated Arctic soils. Bacterial community composition of uncontaminated, diesel-contaminated and diesel+MAP soils was assessed through multiplexed 16S (ribosomal RNA) rRNA gene sequencing on an Ion Torrent Personal Genome Machine, while hydrocarbon degradation was measured by gas chromatography analysis. Diversity of 16S rRNA gene sequences was reduced by diesel, and more so by the combination of diesel and MAP. Actinobacteria dominated uncontaminated soils with <10% organic matter, while Proteobacteria dominated higher-organic matter soils, and this pattern was exaggerated following disturbance. Degradation with and without MAP was predictable by initial bacterial diversity and the abundance of specific assemblages of Betaproteobacteria, respectively. High Betaproteobacteria abundance was positively correlated with high diesel degradation in MAP-treated soils, suggesting this may be an important group to stimulate. The predictability with which bacterial communities respond to these disturbances suggests that costly and time-consuming contaminated site assessments may not be necessary in the future.     

gamma-Glutamyl transpeptidase GGT4 initiates vacuolar degradation of glutathione S-conjugates in Arabidopsis

The xenobiotic monochlorobimane is conjugated to glutathione in the cytosol of Arabidopsis thaliana, transported to the vacuole, and hydrolyzed to cysteine S-bimane [Grzam, A., Tennstedt, P., Clemens, S., Hell, R. and Meyer, A.J. (2006) Vacuolar sequestration of glutathione S-conjugates outcompetes a possible degradation of the glutathione moiety by phytochelatin synthase. FEBS Lett. 580, 6384-6390]. The work here identifies gamma-glutamyl transpeptidase 4 (At4g29210, GGT4) as the first step of vacuolar degradation of glutathione conjugates. Hydrolysis of glutathione S-bimane is blocked in ggt4 null mutants of A. thaliana. Accumulation of glutathione S-bimane in mutants and in wild-type plants treated with the high affinity GGT inhibitor acivicin shows that GGT4 is required to initiate the two step hydrolysis sequence. GGT4:green fluorescent protein fusions were used to demonstrate that GGT4 is localized in the lumen of the vacuole.