Rhodobacter sphaeroides,a novel tumor‐targeting bacteria that emits natural near‐infrared fluorescence

Several optical imaging techniques have been used to monitor bacterial tropisms for cancer. Most such techniques require genetic engineering of the bacteria to express optical reporter genes. This study investigated a novel tumor‐targeting strain of bacteria, Rhodobacter sphaeroides 2.4.1 (R. sphaeroides), which naturally emits near‐infrared fluorescence, thereby facilitating the visualization of bacterial tropisms for cancer. To determine the penetration depth of bacterial fluorescence, various numbers of cells (from 10⁸ to 10¹⁰ CFU) of R. sphaeroides and two types of Escherichia coli, which stably express green fluorescent protein (GFP) or red fluorescent protein (RFP), were injected s.c. or i.m. into mice. Bacterial tropism for cancer was determined after i.v. injection of R. sphaeroides (10⁸ CFU) into mice implanted s.c. with eight types of tumors. The intensity of the fluorescence signal in deep tissue (muscle) from R. sphaeroides was much stronger than from E. coli‐expressing GFP or RFP. The near‐infrared fluorescence signal from R. sphaeroides was visualized clearly in all types of human or murine tumors via accumulation of bacteria. Analyses of C‐reactive protein and procalcitonin concentrations and body weights indicated that i.v. injection of R. sphaeroides does not induce serious systemic immune reactions. This study suggests that R. sphaeroides could be used as a tumor‐targeting microorganism for the selective delivery of drugs to tumor tissues without eliciting a systemic immune reaction and for visualizing tumors.     

球形红细菌厌氧降解邻二氯苯及其机理研究

研究分析光合细菌球形红细菌在厌氧光照条件下降解邻二氯苯的条件和机理.结果表明,在厌氧光照条件下球形红细菌的最佳生长和对邻二氯苯的最佳降解条件为:pH 7.0,温度为30℃,接种量10%.在最佳条件下,邻二氯苯的去除率可达90%以上;其降解中间产物主要有氯苯、4-羟基苯甲酸;根据降解产物的分析,推断球形红细菌降解邻二氯苯的机理主要是按照先脱掉一个氯原子生成氯苯,然后氯苯进一步脱氯并通过4-羟基苯甲酸的代谢途径开环进行.     

Isolation and characterization of transposon Tn5 mutants of Rhodobacter sphaeroides deficient in both nitrate and chlorate reduction.

Abstract The wild-type strain Rhodobacter sphaeroides DSM 158 is a nitrate-reducing bacterium with a periplasmic nitrate reductase. Addition of chlorate to the culture medium causes a stimulation of the phototrophic growth, indicating that this strain is able to use chlorate as an ancillary oxidant. Several mutant strains of R. sphaeroides deficient in nitrate reductase activity were obtained by transposon Tn5 mutagenesis. Mutant strain NR45 exhibited high constitutive nitrate and chlorate reductase activities and phototrophic growth was also increased by the presence of chlorate. In contrast, the stimulation of growth by chlorate was not observed in mutant strains NR8 and NR13, in which transposon Tn5 insertion causes the simultaneous loss of both nitrate and chlorate reductase activities. Tn5 insertion probably does not affect molybdenum metabolism since NR8 and NR13 mutants exhibit both xanthine dehydrogenase and nitrogenase activities. These results that a single enzyme could reduce both nitrate and chlorate in R. sphaeroides DSM 158.     

Damping of oscillations in the semiquinone absorption in reaction centers after successive flashes. Determination of the equilibrium between QA−QB and QAQB−

A quantitative model for the damping of oscillations of the semiquinone absorption after successive light flashes is presented. It is based on the equilibrium between the states Q_A^?Q_B and Q_AQ_B^?. A fit of the model to the experimental results obtained for reaction centers from Rhodopseudomonas sphaeroides gave a value of $\text{α =}\text{[}\text{Q}{}_{\mathrm{<mtext>A</mtext>}}{}^{\mathrm{?}}\text{Q}{}_{\mathrm{<mtext>B</mtext>}}\text{]}\text{([}\text{Q}{}_{\mathrm{<mtext>A</mtext>}}{}^{\mathrm{?}}\text{Q}{}_{\mathrm{<mtext>B</mtext>}}\text{] + [}\text{Q}{}_{\mathrm{<mtext>A</mtext>}}\text{Q}{}_{\mathrm{<mtext>B</mtext>}}{}^{\mathrm{?}}\text{])}\text{= 0.065 ± 0.005}$ (T = 21°C, pH 8).     

念珠藻葛仙米的藻殖段分化——信号转导参与过程的初步研究

钙调素拮抗剂W_7和FPZ均可抑制葛仙米藻殖段的分化。利用FPZ作为荧光染色剂发现 ,经过钙调素拮抗剂处理的藻体有较低的钙调素分布。在达到一定浓度后 ,金属离子螯合剂EGTA也抑制藻殖段分化。金霉素 (CTC)荧光显示 ,经EGTA处理的藻细胞其膜钙分布大大减少。但实验表明 ,藻殖段得以正常分化却是多种金属离子共同参与的结果。一氧化氮 (NO)可以大大降低葛仙米藻殖段的分化率 ,一氧化氮合成酶 (NOS)的抑制剂NNA也可以抑制藻殖段的分化。外源地加入氧自由基抑制了藻殖段的分化 ,但是氧自由基的清除物过氧化氢酶 (CA)也使藻殖段分化完全受抑制。以上结果初步表明 ,葛仙米藻殖段分化是个复杂的、信号转导参与的过程     

球形念珠藻的生化组成分析

测定了葛仙米的生化组成,结果表明,葛仙米的蛋白、氨基酸和总糖含量分别为28．33％、23．35％和24．19％;灰分和粗脂肪原含量分别为14．63％和5．34％;还含有较为丰富的粗脂肪和无机元素,特别是钙、镁元素的含量较高。     

The use of two-dimensional gradient plates in determining the responses of non-sulphur purple bacteria to pH and NaCl concentration

Abstract Chemical gradients can easily be established in agar gels to test behaviour of bacteria in respect to two variables. Agar plates with pH and NaCl gradients have provided a simple technique for useful comparison of heterotrophic bacteria and the present paper extends observations to non-sulphur purple photosynthetic bacteria.     

一株类胡萝卜素高产菌S2的研究

从废水中分离到一株球形红杆菌Ｓ２,能够在光照微氧条件下以葡萄糖为碳源,脲为氮源的培养基上旺盛生长,并产生大量红色胞内类胡萝卜素。在最适合条件下,经３８ｈ分批发酵,类胡萝卜素产量达３１２ｍｇ／Ｌ。     

The stereoisomerism of bacterial, reaction-center-bound carotenoids revisited: An electronic absorption, resonance Raman and H-NMR study

In order to solve discrepancies between earlier assignments we have reinvestigated the stereoisomerism of the spheroidene molecule bound to reaction centers (RC) of Rhodobacter sphaeroides. A stable cis isomer could be extracted and purified from the reaction centres by working at very low ambient light. Resonance Raman spectroscopy showed that this cis isomer assumed the same configuration as that of the RC-bound molecule. Proton-NMR spectroscopy of the extracted isomer permitted to assign it the 15–15′ mono cis configuration. Comparisons between resonance Raman spectra of the native form and of the 15 cis extract showed that, in the reaction center, 15 cis spheroidene is in addition twisted into a non-planar conformation. Comparisons of extraction-induced changes in relative intensities of Raman bands of the 760–1060 cm⁻¹ regions, which largely correspond to out-of-plane modes, further indicated that the out-of-plane twist of RC-bound spheroidene should predominantly affect C₈–C₁₂ and/or C_8′–C_12′ regions of the molecule rather than the central region. Comparisons between difference electronic absorption spectra of RC-bound spheroidene and of RC-bound methoxyneurosporene showed that the out-of-plane twisting of both these native forms results in a drastic weakening of their ¹C ← ¹A electronic transitions, compared with those of the planar, 15 cis forms. Finally, it is proposed, on the basis of their resonance Raman spectra, that spirilloxanthin bound to RCs of Rhodospirillum rubrum as well as dihydroneurosporene or dihydrolycopene bound to RCs of Rhodopseudomonas viridis shares 15 cis configurations and out-of-plane twisting with carotenoids bound to RCs of various strains of Rb. sphaeroides.     

Resolved difference spectra of redox centers involved in photosynthetic electron flow in Rhodopseudomonas capsulata and rhodopseudomonas sphaeroides

1. In Rhodopseudomonas sphaeroides the Q_x absorption band of the reaction center bacteriochlorophyll dimer which bleaches on photo-oxidation is both blue-shifted and has an increased extinction coefficient on solubilisation of the chromatophore membrane with lauryldimethylamine-N-oxide. These effects may be attributable in part to the particle flattening effect.2. The difference spectrum of photo-oxidisable c type cytochrome in the chromatophore was found to have a slightly variable peak position in the α-band (λ_max at 551–551.25 nm); this position was always red-shifted in comparison to that of isolated cytochrome c₂ (λ_max at 549.5 ± 0.5 nm). The shift in wavelength maximum was not due to association with the reaction center protein. A possible heterogeneity in the c-type cytochromes of chromatophores is discussed.3. Flash-induced difference spectra attributed to cytochrome b were resolved at several different redox potentials and in the presence and absence of antimycin. Under most conditions, one major component, cytochrome b₅₀ appeared to be involved. However, in some circumstances, reduction of a component with the spectral characteristics of cytochrome b_?90 was observed.4. Difference spectra attributed to (BChl)₂, Q^?_II, c type cytochrome and cytochrome b₅₀ were resolved in the Soret region for Rhodopseudomonas capsulata.5. A computer-linked kinetic spectrophotometer for obtaining automatically the difference spectra of components functioning in photosynthetic electron transfer chains is described. The system incorporates a novel method for automatically adjusting and holding the photomultiplier supply voltage.