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.     

Benzoate degradation by Rhodococcus opacus 1CP after dormancy: Characterization of dioxygenases involved in the process

The process of benzoate degradation by strain Rhodococcus opacus 1CP after a five-year dormancy was investigated and its peculiarities were revealed. The strain was shown to be capable of growth on benzoate at a concentration of up to 10 g L^?1. The substrate specificity of benzoate dioxygenase (BDO) during the culture growth on a medium with a low (200–250 mg L^?1) and high (4 g L^?1) concentration of benzoate was assessed. BDO of R. opacus 1CP was shown to be an extremely narrow specificity enzyme. Out of 31 substituted benzoates, only with one, 3-chlorobenzoate, its activity was higher than 9% of that of benzoate. Two dioxygenases, catechol 1,2-dioxygenase (Cat 1,2-DO) and protocatechuate 3,4-dioxygenase (PCA 3,4-DO), were identified in a cell-free extract, purified and characterized. The substrate specificity of Cat 1,2-DO isolated from cells of strain 1CP after the dormancy was found to differ significantly from that of Cat 1,2-DO isolated earlier from cells of this strain grown on benzoate. By its substrate specificity, the described Cat 1,2-DO was close to the Cat 1,2-DO from strain 1CP grown on 4-methylbenzoate. Neither activity nor inhibition by protocatechuate was observed during the reaction of Cat 1,2-DO with catechol, and catechol had no inhibitory effect on the reaction of PCA 3,4-DO with protocatechuate.     

不同硫源对红串红球菌生长和脱硫比活性的效应

本文牛磺酸、硫酸镁和二苯噻吩（ＤＢＴ）分别作为唯一硫源以及ＤＢＴ不同供给量对红串红球菌生长和脱硫比活性的效应进行了研究,结果表明,红串红球菌肯 利用有机和无机硫化合物作为硫源的能力。与牛磺酸和到镁相比,ＤＢＴ是促进红串红球菌生长和提高其脱硫比活红串红球菌的最大脱硫比活性达７９．１ｍｍｏｌ２ＨＢＰ．ｋｇ＾－１．ｈ＾－１。影响红串红球菌脱硫比活性的因素主要是其生长的不同阶段、ＤＢＴ供给量和羟基二苯生产     

厌氧后猪场废水资源化制备微生物絮凝剂

通过检测碳源、氮源、磷酸盐和pH等对微生物絮凝剂絮凝率的影响,考察了红平红球菌利用厌氧处理后的猪场废水生产微生物絮凝剂的性能。通过正交实验研究各因素对絮凝率的交互影响以及影响程度的相对大小,从而确定了最佳培养基。结果表明,1.0 L厌氧处理后的猪场废水中加入蔗糖10.0 g、KH2PO42.0 g、K2HPO45.0 g、NaCl 1.0 g、Mg-SO40.2 g,在温度30℃、摇床速度120 r/min、pH=8的条件下发酵72 h后,发酵液的絮凝率可达到92.57%。表明厌氧处理后的猪场废水中加入适量的蔗糖和磷酸盐,完全可以作为一种有效的生产微生物絮凝剂的培养基。     

红球菌跨膜运输荧蒽的差异膜蛋白分析

提取不同时间荧蒽诱导下红球菌BAP-1分泌的膜蛋白,利用同位素标记相对和绝对定量技术（iTRAQ技术）结合液相二级质谱对差异膜蛋白进行聚类以及生物信息学分析.旨在从蛋白质层面上研究红球菌BAP-1跨膜运输荧蒽过程中起关键作用的膜蛋白种类及其作用机制.结果共鉴定到172个差异膜蛋白.通过差异膜蛋白的COG和GO功能分析,发现荧蒽的加入主要影响了红球菌BAP-1细胞膜上与能量、转运相关的膜蛋白.ABC转运蛋白和TonB依赖转运蛋白作为微生物主要的载体蛋白对BAP-1跨膜运输荧蒽起到了重要作用.过氧化氢酶和超氧化物歧化酶在第6d有一个显著上调,作为抗氧化防御机制来保护微生物.各种膜蛋白在不同阶段发挥着各自作用,这些蛋白共同组成蛋白互作网络调控红球菌BAP-1的跨膜运输过程.     

降解菌djl-6多菌灵水解酶的提取及性质

以往对多菌灵降解菌Rhodococcus qingshengii sp.nov.djl-6的降解途径研究显示,该菌株首先通过多菌灵水解酶将多菌灵水解成二氨基苯并咪唑,从而对多菌灵进行脱毒.为开发酶制剂并有效应用于环境中残留污染物多菌灵的降解,比较了不同提取方法(高压细胞破碎、超声波破碎和添加溶菌酶破碎)对多菌灵水解酶提取效率的影响,并对其酶学特性进行了初步研究.结果表明,djl-6菌株在LB培养基中培养72~84 h,生长量和产酶量均达到最大值.采用超声波破碎提取酶的效率较高(蛋白浓度为7.92 mg/mL),但酶活损失较大(比酶活只有1.2 U/μg protein).多菌灵水解酶属于一种胞内组成型酶.该酶水解多菌灵的最适pH值为7.0,最适温度为30℃,Zn2+和K+对酶活有一定的抑制作用.     

Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

鸡粪除臭菌的筛选、培养条件优化及其应用

为了筛选出具有高效除臭能力的菌株,采用驯化富集、平板划线的方法从鸡粪中筛选除臭微生物,采用初筛、复筛相结合的方法检测所筛选微生物的除臭效果。初筛采用嗅阈值法,复筛以氨气和硫化氢的去除率为检测指标,最终获得除臭效果最好的菌株CCJZ022。经过形态学观察、生理生化鉴定和16S rRNA基因序列分析,确定菌株CCJZ022为红平红球菌(Rhodococcus erythropolis)。对其初始pH值、培养温度及接种量进行优化,发现该菌株在初始pH值为7.0、培养温度为30℃、接种量为12%时除臭效果最好,对氨气和硫化氢的去除率分别可达66.73%和54.51%。利用该菌株进行鸡粪除臭的小试也取得了很好的效果,表明该菌株可以应用于鸡粪除臭。     

生物脱硫技术用于清洁燃料生产的初步研究

从高硫油田筛选的红球菌Rhodococcus sp．FS-2可以通过专一性断裂C-S键的“4S”途径将二苯并噻吩（DBT）转化成2-羟基联苯，从而降低油品中的硫含量。利用该菌株对DBT和汽油、柴油的脱硫研究结果表明，FS-2菌株对DBT及柴油中的有机硫有良好的选择性脱除作用，而且脱硫前后的烃类组分基本没有改变。说明该菌的脱硫过程不会破坏燃料油的有效成分，可以用于燃料油的深度脱硫。