Purification and Properties of a Poly (β-hydroxybutyrate) Depolymerase From Penicillium sp.

An extracellular poly (β-hydroxybutyrate) (PHB) depolymerase was purified from a Penicillium sp. DS9701-09a by centrifugation, ultrafiltration, precipitation and gel filtration chromatography. The specific activity of the purified enzyme was 37.9-folds higher than that of the culture supernatant and the recovery yield was 11.8%. The PHB deploymerase molecular mass was 44.8 kDa from analysis of both Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Matrix-assisted laser desorption-time-of-flight (MALDI-TOF) mass spectrometer. The isoelectric point of 6.7 for the enzyme was determined by a two-dimensional electrophoresis. The optimum enzyme activity was observed at a temperature of 50 °C and pH 5.0. The apparent K _m of the enzyme was found to be 1.35 mg/mL. The PHB depolymerase consisted of 16 kinds of normal amino acids. The secondary structure of the enzyme was determined by CD spectrum. α-helix and β-turn were found to be 66% and 34% for the enzyme without ammonium sulphite. Chemical inhibition on the PHB depolymerase activity was examined and EDTA was found to have a significantly inhibitory effect.     

Promotion of hexadecyltrimethyleamine bromide to the damage of Alexandrium sp. LC3 by cupric glutamate

The effect of hexadecyltrimethyleamine bromide （HDTMAB） on the removal of A lexandrium sp. LC3 under cupric glutamate stress was investigated. Toxic effect of cupric glutamate on A lexandrium sp. LC3 was significantly promoted in the presence of HDTMAB, especially at 3.0 cmc of HDTMAB. It was found that the sulfhydryl group content of the cell decreased, while the malonaldehyde content and membrane permeability increased when A lexandrium sp. LC3 was treated with HDTMAB and cupric glutamate complex, compared with cupric glutamate alone. The data suggest that HDTMAB might stimulate the damage of A lexandrium sp. LC3 by enhancing the membrane permeability.     

芽孢杆菌发酵产碱性果胶酶温度控制策略

采用自行筛选获得一株产碱性果胶酶芽孢杆菌WSH03-09,在小型发酵罐中研究了不同温度对碱性果胶酶分批发酵的影响,结果表明,在恒定39℃条件下,可获得最高酶活5．39u／mL,各温度条件下的菌体干重相差不多,最终均能达11．5g／L左右;在发酵前期,控制温度41℃时最有利于菌体的生长,而在产物合成期,控制37℃有利于获得较高的产物合成比速,在此基础上,提出分阶段温度控制策略,采用此温度控制策略进行碱性果胶酶的发酵,碱性果胶酶酶活达5．99u／mL,比采用单一温度下的最大值提高了11％,其它各项指标也有较大提高．图6表1参6     

1株耐盐异养硝化-好氧反硝化菌Zobellella sp.B307的分离及脱氮特性

从胶州湾沉积物中分离出1株异养硝化-好氧反硝化菌株B307,采用16S rRNA基因序列分析对该菌株进行鉴定,采用单因素实验对其进行条件优化和耐盐特性研究,并在最优条件下考察其在单一和混合氮源中的脱氮效果.结果表明,该菌为Zobellella sp.,其最佳碳源为丁二酸钠,最适C/N为5,最适初始p H为9,最适温度为35~40℃.该菌株在混合氮源体系中12 h对NH_4~+-N和NO_3~--N的去除率分别为98.35%和99.75%;在盐度为75 g·L~(-1)(以NaCl计)条件下24 h对NH_4~+-N和NO_3~--N去除率仍分别保持在97.67%和94.39%.表明该菌株具有高效的异养硝化-好氧反硝化能力和较强的耐盐特性,在高盐废水脱氮等领域具有广泛的应用前景.     

A bioaugmentation failure caused by phage infection and weak biofilm formation ability

Two biological aerated filters (BAF) were setup for ammonia removal treatment of the circulation water in a marine aquaculture. One of the BAFs was bioaugmented with a heterotrophic nitrifying bacterium, Lutimonas sp. H10, where the ammonia removal was not improved and the massive inoculation was even followed by a nitrification breakdown from day 9 to 18. The nitrification was remained stable in control BAF operated under the same conditions. Fluorescent in situ hybridization (FISH) with rRNA-targeted probes and cultivable method revealed that Lutimonas sp. H10 almost disappeared from the bioaugomented BAF within 3 d, and this was mainly due to the infection of a specific phage as revealed by flask experiment, plaque assay and transmission electron observation. Analyses of 16S rRNA gene libraries showed that bacterial groups from two reactors evolved di erently and an overgrowth of protozoa was observed in the bioaugmented BAF. Therefore, phage infection and poor biofilm forming ability of the inoculated strain are the main reasons for bioaugmentation failure. In addition, grazing by protozoa of the bacteria might be the reason for the nitrification breakdown in bioaugmented BAF during day 9–18.     

Isolation of Cr(VI) reducing bacteria from industrial e uents and their potential use in bioremediation of chromium containing wastewater

The present study is aimed at assessing the ability of Bacillus sp.JDM-2-1 and Staphylococcus capitis to reduce hexavalent chromium into its trivalent form.Bacillus sp.JDM-2-1 could tolerate Cr(Ⅵ) (4800 μg/mL) and S.capitis could tolerate Cr(Ⅵ) (2800 μg/mL).Both organisms were able to resist Cd2+ (50 μg/mL),Cu2+ (200 μg/mL),Pb2+ (800 μg/mL),Hg2+ (50 μg/mL) and Ni2+ (4000 μg/mL).S.capitis resisted Zn2+ at 700 μg/mL while Bacillus sp.JDM-2-1 only showed resistance up to 50 μg/mL.Bacillus sp.JDM-2-1 and S.capitis showed optimum growth at pH 6 and 7,respectively,while both bacteria showed optimum growth at 37℃.Bacillus sp.JDM-2-1 and S.capitis could reduce 85% and 81% of hexavalent chromium from the medium after 96 h and were also capable of reducing hexavalent chromium 86% and 89%,respectively,from the industrial effluents after 144 h.Cell free extracts of Bacillus sp.JDM-2-1 and S.capitis showed reduction of 83% and 70% at concentration of 10 μg Cr(Ⅵ)/mL,respectively.The presence of an induced protein having molecular weight around 25 kDa in the presence of chromium points out a possible role of this protein in chromium reduction.The bacterial isolates can be exploited for bioremediation of hexavalent chromium containing wastes,since they seem to have the potential to reduce the toxic hexavalent form to its nontoxic trivalent form.     

一株高效十四烷降解菌的筛选及降解条件优化

采集某炼油厂废水处理车间的回流污泥,以十四烷作为唯一碳源筛选分离到一株高效十四烷降解菌。经形态学观察和生理生化特征研究,鉴定为布兰汉氏球菌(Branhamella sp.)。对其降解十四烷的条件进行了优化,实验结果表明,最适降解条件为接种量0.5%,温度35℃,初始pH=7,碳氮摩尔比为0.4:1,在最佳条件下,当十四烷的初始浓度约为50mg/L时,在12h内降解率高达98%以上。     

4株邻苯二甲酸二丁酯降解菌的分离鉴定及其相关降解基因的克隆

从土壤中分离纯化出4株能降解邻苯二甲酸二丁酯(DBP)的菌株,分别命名为JDC-1、JDC-8、JDC-9、JDC-12,并对其进行了形态学、生理生化及分子生物学鉴定.菌株革兰氏染色阳性,16S rDNA序列分析显示4株菌均与节杆菌属(Arthrobacter sp.)有99%以上的序列相似性,初步判断这4株菌为Arthrobacter sp..通过PCR扩增及克隆,均获得了1个约900 bp的DNA片段,测序结果显示该片段与Arthrobacter keyseri的邻苯二甲酸3,4-双加氧酶基因的核苷酸序列相似性为96%以上.对4株菌的最适生长条件及对DBP的降解能力进行了分析,结果显示,4株菌的最适生长条件为pH 7.0~8.5,温度30~35℃.以DBP作为目标测试物,在适宜条件下测试了4株菌的降解能力,显示这4株菌均为高效降解菌,效率最高的JDC-1能在28 h内将500 mg/L的DBP降解完全,最慢的JDC-8经40 h能将500 mg/L的DBP降解完全,本研究对于DBP降解机制的研究及微生物资源的开发都具有重要意义.     

黄单胞菌生物滴滤塔处理硫化氢能力及产物研究

以黄单胞菌为硫氧化菌种挂膜于生物滴滤塔,研究其去除能力及产物.通过测定压降、生物量确定挂膜时间,以进出气口浓度变化反映去除效率,测定营养液pH值变化,用SEM电镜扫描、能谱分析、X射线衍射对产物进行分析.结果表明,黄单胞菌挂膜时间为4 d,持续运行,菌种均匀地分布于填料表面;保持气体流量为1 L/min,H_2S气体质量浓度在2 000 mg/m~3以下时,去除率大于90.5%,单位体积最大生化去除量为43.23 g/(m~3·h);以10 d为周期添加营养,系统的pH值在5～7;自然沉降得到的黄色颗粒状沉淀粒径为3.161～31.90 μm;能谱分析表面主要成分为C、O、S,应该为微生物和S系物;X射线衍射分析表明干燥粉末的主要无机物质存在形态为S单质.塔在弱酸环境运行,能减少对装置的腐蚀,营养液中通过物理沉降、过滤、低速离心的简单手段,就能回收产物S单质.     

DMF降解菌MBYD-1的分离、鉴定及降解特性

从农药厂污水处理池的活性污泥中分离得到一株能以二甲基甲酰胺(DMF)为唯一碳、氮源生长的菌株MBYD-1.经形态观察、生理生化实验及16S rRNA基因序列分析,将菌株初步鉴定为假单胞菌属 (Pseudomonas sp.).菌株MBYD-1降解DMF的最适条件为:pH 7.0,30℃.当DMF浓度为400mg/L,接种量为2%时,菌株能在72h内将DMF完全矿化.底物广谱性实验表明,菌株MBYD-1能在甲酸盐和甲醛中很好地生长,在甲酰胺中的生长情况优于二甲胺.