Effects of glucose dehydrogenase gene knock-out in Acetobacter sp. On the metabolism of carbon sources and the synthesis of 3-hydroxypropionic acid北大核心CSCD

3-Hydroxypropionic acid (3-HP) is an emerging platform chemical with a high added-value. Resting cells of Acetobacter sp. can efficiently catalyze 1,3-propanediol (1,3-PDO) to 3-HP. Glucose is oxidized by the membrane-bound dehydrogenase, resulting in an acidic environment that inhibits cell growth and reduces the biomass. We deleted the gdh gene for glucose dehydrogenase (GDH), and investigated the effects on cell growth, carbon metabolism, and 3-HP production. The gdh gene knocked-out showed a 1.72-fold increase in biomass in the mixed medium containing glucose and glycerol. A carbon flux analysis showed that glucose was converted to gluconic acid by GDH, followed by an oxidation to 2-ketogluconic acid. In addition, a small percentage of the gluconic acid was degraded via the pentose phosphate pathway. Glycerol was phosphorylated and entered the central pathway (gluconeogenesis). Results indicate that the deletion of gdh can effectively promote higher cell densities and improve the catalytic performance for the production of 3-HP, and thus provide a theoretical reference for improving the carbon source utilization and the catalytic performance of acetic acid bacteria. © 2018 Science Press. All rights reserved.     

Isolation,identification, and the degradation characteristics of a thiobencarb-degrading bacterium Bacillus sp. T2北大核心CSCD

Thiobencarb, a thiocarbamate herbicide, is widely used to control weeds in rice paddies. Screening for highly efficient thiobencarb-degrading bacteria is important for the bioremediation of thiobencarb-contaminated environments. The aim of this study was to isolate and identify highly efficient thiobencarb-degrading bacteria and to identify the degradation pathway and the degrading properties. The thiobencarb-degrading strain was isolated using methods of microbiological acclimation and enrichment and was then identified using a 16S rRNA phylogenetic analysis. The degrading properties of the isolated bacterium were determined by single-factor experiments, and the degradation products were identified using gas chromatography-mass spectrometry (GC-MS). A thiobencarb-degrading strain T2, which can utilize thiobencarb as the sole source of carbon for energy and growth, was isolated from paddy soil. Strain T2 degraded more than 98.3% of 0.4 mmol/L of thiobencarb within 36 h. It was preliminarily identified as Bacillus sp. T2 according to the 16S rRNA gene analysis and from its morphological, physiological, and biochemical characteristics. The metabolic products of the thiobencarb degradation for strain T2 were identified as 4-chlorobenzyl mercaptan, 4-chlorobenzaldehyde, and 4-chlorobenzoic acid by the GC-MS. Based on metabolite identification, it was speculated that thiobencarb degradation in strain T2 was initiated by the hydrolysis of the thioester bond to produce 4-chlorobenzyl mercaptan, which was further oxidized to 4-chlorobenzaldehyde and 4-chlorobenzoic acid. The thiobencarb degradation that was initiated by the hydrolysis of the thioester bond by strain T2 is a new metabolic pathway, which provides valuable research material and reliable experimental data for revealing the metabolic process and mechanism of thiobencarb microbial degradation in soil. The strain Bacillus sp. T2 has a very high degradation efficiency, suggesting it is a good prospect for microbial remediation in thiobencarb-polluted environments. © 2018 Science Press. All rights reserved.     

红球菌-R04生物降解多卤代联苯的影响因素研究

研究了不同取代元素以及取代元素的数量、位置等因素对 Rhodococcus sp. R04降解多卤代联苯的影响.结果表明,联苯苯环第4位上的氢分别被氟、氯、溴以及甲基基团取代后降解率由高到低依次为4-FB>4-CB34-MB>4-BrB;当取代元素相同时,随着取代原子数目的增加,多卤代联苯降解率降低;当取代元素及取代原子数目相同时,不同位置的取代对卤代联苯的降解影响较大,特别是2,6位取代会强烈抑制关键酶对卤代联苯的催化降解,导致降解率降低.     

Isolation,identification, and acetochlor-degrading potential of a novel Rhodococcus sp. MZ-3

A new species of Rhodococcus, designated strain MZ-3, which could degrade acetochlor efficiently were isolated and identified. The isolate could degrade and utilize acetochlor as the sole source of carbon, nitrogen, and energy for growth. The optimal conditions for the degradation and growth of MZ-3 were pH 7.0 and 30°C. Under these conditions, this strain could completely degrade 200 mg/L of acetochlor within 12 h of incubation. During the biodegradation process, the enantioselectivity of the strain was investigated using a chiral high-performance liquid chromatography (HPLC) system. However, no obvious enantioselectivities were found. 2-chloro-N-(2-methyl-6-ethylphenyl) acetamide (CMEPA) was detected as the intermediate using liquid chromatography-mass spectrometry (LC-MS) analyses. Our results suggest that strain MZ-3 might be a promising microorganism for the bioremediation of acetochlor-contaminated environments because of its acetochlor-degrading performance.     

好氧砷还原菌对吸附态砷迁移转化的影响

利用静态和动态土柱实验初步研究了好氧砷还原菌Bacillus sp.SXB在砷迁移转化过程中的作用。结果表明,在静态实验中,菌株SXB可以有效还原吸附在针铁矿上的五价砷;在动态土柱实验中,由于原土中砷是以三价砷形式存在,因此,不论是空白对照还是菌株SXB加入处理,出水中砷均为三价砷;当孔隙体积大于100时,铁的溶出开始增加。菌株SXB的加入,轻微地促进了土壤中砷的释放,而对铁的释放并没有明显影响。     

两株氢噬胞菌的萘降解特性分析

对两株分离自内蒙古乌梁素海的氢噬胞菌X32和X12的培养条件和萘降解特性进行研究。实验结果表明:菌株X32和X12的最适生长pH为7.0,最适生长温度为30~35℃,最适盐度w(NaCl)为1%;当初始萘质量浓度为3 500 mg/L时,对数生长期的菌株X32对萘的降解活性可达53.9 nmol/(mg·min),而菌株X12可达34.8 nmol/(mg·min);菌株X32在培养48 h后进入稳定期,60 h时萘降解率达91.43%;菌株X12在培养60 h后进入稳定期,90 h时萘降解率达93.93%。氢噬胞菌X32和X12是两株具有较高应用价值的多环芳烃降解细菌。     

Performance evaluation of isoproturon-degrading indigenous bacterial isolates in soil microcosm

Isoproturon (IPU)-degrading soil bacteria were isolated from herbicide-applied wheat fields. These isolates were identified using cultural, morphological, biochemical and 16S rRNA sequencing methods. 16S rRNA sequences of both the bacterial isolates were compared with NCBI GenBank data base and identified as Bacillus pumilus and Pseudoxanthomonas sp. A soil microcosm study was carried out for 40 days in six different treatments. Experimental results revealed maximum 95.98% IPU degradation in treatment 6 where bacterial consortia were augmented in natural soil, followed by 91.53% in treatment 5 enriched with organic manure as an additional carbon source. However, only 14.03% IPU was degraded in treatment 1 (control) after 40 days. In treatments (2–4), 75.59%, 70.92% and 77.32% IPU degradation was recorded, respectively. IPU degradation in all the treatments varied significantly over the control. 4-Isopropylaniline was detected as IPU degradation by-product in the medium. The study confirmed that B. pumilus and Pseudoxanthomonas sp. performed effectively in soil microcosms and could be employed profitably for field-scale bioremediation experiments.     

滏阳河河流水体中重金属污染特征及其对青海弧菌和斜生栅藻的毒性效应

为调查滏阳河水系的重金属污染状况,研究河流重金属污染对水生生物的毒性,根据河流结构、水文条件、排污口分布并考虑空间分布的均匀性,选取66个采样位点,采集河水及对应的表层沉积物样品,分析了样品中的重金属含量。用斜生栅藻和青海弧菌Q67作为模式生物,根据滏阳河水重金属污染较严重的邯郸近郊2号采样点采集水样的重金属含量配制系列重金属浓度的模拟河水进行重金属污染河水的生态毒性测试。结果表明,在全部66个采样点中,29个采样点河水重金属含量超过国家地表水III类水体重金属含量标准,主要污染元素是Hg、Pb、Cr、Zn。几乎所有采样点河水Mn和Fe含量都大大高于国家集中式生活饮用水地表水源地补充项目规定的标准限值。根据2号采样点河水样品中的重金属含量配成的模拟河水对青海弧菌Q67的EC₅₀值为6.65%,为毒性极强的污染物。模拟河水样品对斜生栅藻的抑制作用较小,在实验的最高浓度下(1000倍河水重金属含量)暴露4 d尚未引起半数藻细胞死亡。随河水重金属浓度上升,斜生栅藻超氧化物歧化酶(SOD)活性总体上呈现先升高后降低的趋势,丙二醛(MDA)含量变化则与此相反,反映河水重金属污染可引起藻细胞的氧化损伤。叶绿素a和b含量则随暴露浓度的提高逐渐降低。在重金属浓度达到2号采样点河水的10%时,斜生栅藻叶绿素a含量已有显著降低,MDA含量显著升高,海河流域重金属污染对生态系统的影响应予以重视。河水发光菌Q67的生长抑制率、斜生栅藻的叶绿素a和MDA含量可以作为评估河流重金属污染生态危害的指标。     

氯氰菊酯降解菌CY22-7的分离鉴定及降解特性研究

从采集的多种土壤样品中.获得了19株在3种筛选平板上均生长良好的候选氯氰菊酯降解菌.通过生理生化特性鉴定和16S rDNA序列的测定及比对.将其中的氯氰菊酯降解菌CY22-7鉴定为中华根瘤菌属(Sinorhizobium sp.).同时,对氯氰菊酯降解菌CY22-7的降解特性研究结果表明:(1)氯氰菊酯降解菌CY22-7以5%(体积分数)的接种鼍接种到氯氰菊酯起始质量浓度为100 mg/L的氯氰菊酯乙醇培养基后,于200 r/min、30℃摇床中培养.经过6 d的培养,氯氰菊酯降解菌CY22-7降解了约60%的氯氰菊酯.(2)加人外源营养物质有利于促进氯氰菊酯的降解.其中,葡萄糖和酵母提取物的促进作用最为明显.(3)氯氰菊酯降解菌CY22-7降解氯氰菊酯的最适温度为30℃,最佳pH为6.0.     

微生物絮凝剂产生菌的筛选及其絮凝特性

采用常规的微生物学方法,从垃圾渗滤液中筛选到一株高效微生物絮凝剂产生菌LB1,根据其形态学和生理生化特征初步鉴定该菌株属于假单胞菌属,命名为Pseudonomas sp. LB1。LB1所产微生物絮凝剂的絮凝特性的研究结果表明,LB1的最佳产絮凝时间为96 h,所产絮凝活性物质主要是其生长过程中的胞外分泌物和细胞生长后期的次级代谢产物,菌体细胞本身具有一定的助凝作用。LB1所产絮凝剂最佳加样量为3％（体积分数V/V）;对pH的适应范围较宽,在4～10之间具有较高的絮凝活性,均大于75％;温度为25℃时絮凝率为86.3％,在25～60℃之间,絮凝率基本保持稳定;LB1所产絮凝剂对几种废水具有较好的絮凝效果。