Classification of methylotrophic bacteria according to 5S ribosomal RNA sequencing

5S ribosomal RNA sequences of 33 strains of methylotrophic bacteria were determined. Tentative phylogenetic tree was constructed using the maximum topological similarity principle. Strains under study can be divided into 7 separate branches consistently with the current classification of methylotrophic bacteria. More extensive tree was also built to show the position of methylotrophic bacteria with respect to non-methylotrophic ones. One can conclude that the in contrast to obligate methane-oxidizing bacteria, facultative methylotrophic bacteria do not comprise phylogenetically separate domain on the tree.     

One-carbon compounds transport studies in the obligate methylotroph

Cells of obligate methylotrophic Gram-negative bacterium Methylobacillus flagellatum KT which can only grow on methanol and methylamine media possess three different carriers mediating uptake of methylamin depending on growth conditions. All three uptake systems are energy-dependent, the methylamine uptake was inhibited by oxidative phosphorylation uncoupler and respiratory inhibitors. The first active transport system for methanol in the cells of obligate methylotroph was also demonstrated. The parameters of this system were measured, their dependence on energy, presence of respiratory inhibitors and uncoupler was shown.Abbreviations CCCP Carbonyl cyanide p-(trichloromethoxy)-phenylhydrazone - DCCD N,N-dicyclohexyl-carbodiimide     

Transformation of a new Gram-positive methylotroph,Brevibacterium methylicum,by plasmid DNA

Summary Brevibacterium methylicum is a newly isolated Gram-positive facultatively methylotrophic bacterium that uses the NAD⁺-dependent methanol dehydrogenase for methanol oxidation and assimilates its carbon via the ribulose monophosphate cycle. Protoplasts prepared by lysozyme treatment of B. methylicum cells grown in the presence of glycine were transformed by plasmid shuttle vectors pCEM500 (16.5 kb; Sm^r/Sp^r, Km^r/Gm^r) and pEC71 (7.1 kb; Km^r/Nm^r) constructed on the basis of B. lactofermentum plasmid pAM330 and replicating in Escherichia coli and in amino-acid-producing coryneform bacteria. The resistance markers were found to be expressed in B. methylicum and autonomous plasmid DNAs of various size were isolated from the transformants. The presence of the pAM330 replicon in these plasmids was demonstrated by DNA-DNA hybridization experiments. Offprint requests to: J. Nevera     

Oxidative phosphorylation in membrane vesicles of a gram-positive methylotroph

Membrane preparations, capable of high rates of respiration-linked ATP synthesis, have been obtained from a gram-positive methylotrophic bacterium Bacillus sp. MGA3. NADH, succinate, reduced TMPD and methanol were shown to be suitable substrates for the oxidative phosphorylation. Esterification of orthophosphate was dependent on electron transfer, as evidenced by the requirement for both substrate and oxygen. Phosphorylation was also dependent on ADP and was destroyed by boiling the membrane preparation. The phosphorylation was markedly uncoupled by carbonyl cyanide p-(trichloromethoxy)-phenylhydrazone (CCCP) and was inhibited by N,N-dicyclohexylcarbodiimide (DCCD). KCN caused strong inhibition of substrate oxidation as well as phosphorylation for all substrates tested. Rotenone, amytal and antimycin A caused inhibition when NADH or methanol were used as substrates. Antimycin A inhibited respiration and ATP synthesis with succinate as substrate and had no effect on ascorbate —N,N,N,N-tetramethyl-p-phenylenediimide (TMPD) oxidation by membrane preparations of Bacillus sp. MGA3. P/O ratios determined were 2.4 with NADH, 1.7 with succinate and 0.8 with reduced TMPD. The measured P/O ratio with methanol-oxidizing system was similar to that with NADH (about 2.4).Abbreviations CCCP Carbonyl cyanide p-(trichloromethoxy)-phenylhydrazone - DCCD N,N-dicyclohexylcarbodiimide - TMPD N,N,N,N-tetramethyl-p-phenylenediimide - Q ubiquinone Q     

Enzyme activity of the formate hydrogenlyase complex in Citrobacter freundii

Citrobacter freundii 62 can grow in the absence of oxygen in media containing glucose, peptone, fumarate or malate. When the medium contained fumarate or malate, the culture could grow under anaerobic conditions only in the presence of molecular hydrogen, formate or nitrate. The highest activity of formatehydrogenlyase and hydrogenase was found when C. freundii grew in a medium with glucose and formate. The activity was lower in media with other organic substrates, particularly, in the absence of formate or H2. The activity of hydrogenase was very low in cells grown under aerobic conditions or in the presence of nitrates while the activity of formatehydrogenlyase was not found at all for all practical purposes. The activity of formate dehydrogenase assessed in the presence of methylene blue was rather high irrespective of the conditions under which the culture was grown. However, when the activity of formate dehydrogenase was determined in the presence of benzyl viologen, it was high only in cells grown in the medium with glucose and formate.     

Taxonomic studies on methylotrophic bacteria by 5S ribosomal RNA sequencing

Nucleotide sequences of 5S ribosomal RNA (rRNA) isolated from 19 strains of Gram-negative methylotrophic bacteria were determined. Comparison of these sequences allowed construction of a tentative phylogenetic tree and showed that the bacteria analysed belong to the Proteobacteria and fell into several clusters, including obligate methanotrophs, obligate methylotrophs and several groups of facultative methylotrophs. Taxonomic relations between methylotrophic and non-methylotrophic bacteria are discussed, and the polyphyletic nature of methylotrophy as a taxonomic feature is highlighted.     

正常小鼠高频心电图时域值和功率谱的研究

本文用南京新博公司生产的ＮＨＥ－１０００型心电高频信息检测分析仪研究了正常小鼠（昆明种）高频心电图（ＨＦ－ＥＣＧ）的时域值和ＱＲＳ波群的功率谱。主要结果如下（以正导为例,－Ｘ±ＳＤ）：心率６０３±８８次／ｍｉｎ（ｎ＝７４）;Ｐ－Ｒ间期相对较长。为３４．９±４．７ｍｓ（ｎ＝５８）,占心动周期的３４．９±４．９％,这与人类有很大的不同;ＱＲＳ波宽９．２±１．２ｍｓ,占心动周期的９．２±１．４％（ｎ＝７４）,这一结果与以前的文献报道相差较大。Ｔ波宽１０．３±３．２ｍｓ,占心动周期的１０．３±３．２％;Ｑ－Ｔ间期１９．４±３．２ｍｓ,占心动周期的１９．５±３．６％;ＱＲＳ波群峰－峰值（Ｖｐ－ｐ）为１．４５６±０．４８０ｍＶ;Ｔ波高０．３３６±０．１１５ｍＶ;７３只动物Ⅱ导联高频切迹总数只有３个,扭挫２６个。Ⅱ导联ＱＲＳ波群的功率谱特点：０—８０Ｈｚ的相对能量为４５．４８±１５．３２％;８０—２００Ｈｚ为４３．９７±９．９５％;２００—３００Ｈｚ为８．８９±７．３８％;３００—１０００Ｈｚ为１．６６±２．７４％。     

Integrated membrane systems for gas separation in biotechnology: potential and prospects

Integrated non-porous membrane systems were applied for microbial combustible gas separation processes. Methane/CO₂ mixtures of various concentrations from methane fermentation processes (biogas) were separated using a membrane-separation complex of permabsorber type into individual components of technical grade (more than 95% purity). In experiments with three-component mixtures, using a selective membrane valve with various liquid carriers, all the gases of interest (H₂, CH₄ and CO₂) were obtained at greater than 90% purity in one separation step. The perspectives for the further application of non-porous membrane separating devices for various gaseous mixtures from different microbial processes are discussed.V. Teplyakov and E. Sostina are with the A.V. Topchiev institute of Petrochemical Synthesis, Russian Academy of Sciences, Membrane Research Center, Moscow 117912, Russia. E. Sostina is also, and A. Netrusov is with the Microbiology Department, Moscow University, Moscow 119899, Russia. I. Beckman is with the Chemistry Department, Moscow University, Moscow 119899, Russia.     

17个建兰品种光合特性分析

以17个建兰(Cymbidium ensifolium)品种为材料,采用改良的丙酮法提取叶绿素,再通过Arnon丙酮法公式计算光合色素含量,利用捷克FluorCam开放式叶绿素荧光仪测定不同品种的叶绿素荧光参数。结果表明,17个建兰品种的光合色素和叶绿素荧光参数具有不同程度的差异,其中‘铁骨素’(C. ensifolium ‘Tiegusu’)、‘逸红双娇’(C. ensifolium ‘Yihongshuangjiao’)和‘闽南黄蝶’(C. ensifolium ‘Minnanhuangdie’)的光合色素含量高于其他品种,表明这3个品种具有良好的光合效率,吸收光能的能力较强;‘铁骨素’最大荧光产量(Fm)、Kautsky诱导效应最大荧光(Fp)、PS Ⅱ原初光能转化效率(Fv/Fm)和非光化荧光淬灭系数(NPQ)均为最高。综上可知,‘铁骨素’的光合生理特性优于其他品种,可作为优良建兰品种进行种植推广。