Vitamin B12-dependent methionine synthesis in Rhizobium meliloti

Methionine, among the various additions to the medium, could only replace cobalt ion or vitamin B₁₂ required for the growth of $\text{Rhizobium meliloti}$. It was demonstrated that there exists a vitamin B₁₂-dependent terminal step in the methionine synthesis, that is, N⁵CH₃-tetrahydrofolate-homocysteine transmethylase, which can also catalyze the methyl transfer from CH₃B₁₂ to homocysteine, in the cell-free extracts of Rhizobium meliloti. These facts seem to indicate that the vitamin B₁₂-dependent pathway to methionine functions mainly among the B₁₂-dependent enzymatic systems in the wild-type symbionts and this is the chief nutritional significance of cobalt.     

Kinetics of photosynthetic membrane protein assembly in Rhodopseudomonas spheroides

Glycogen phosphorylase in cell-free extracts of Neurospora crassa is activated 10- to 15-fold by incubation with MgATP^2?. When the MgATP^2? is removed, the active form (a form) reverts to the inactive form (b form). The inactivation requires Mg²⁺ and is inhibited by NaF. The results confirm that Neurospora crassa glycogen phosphorylase exists in two interconvertible forms and strongly suggests that the interconversion is catalyzed by a kinase and phosphatase. The a form was partially purified. The enzyme has a molecular weight of 320,000. Uridine diphosphate glucose is a linear competitive inhibitor with respect to glucose-1-phosphate and a linear non-competitive inhibitor with respect to glycogen. Glucose-6-phosphate is a hyperbolic (partial) noncompetitive inhibitor with respect to all substrates in both directions. The b form of the enzyme in crude cell-free extracts is stimulated 2- to 3-fold by 5′-AMP. As the b form is purified, the 5′-AMP activation is diminished. The molecular weight of the partially purified “b” form was also 320,000.     

Electron acceptors in photosynthetic reaction centers from Rhodopseudomonas spheroides]

Using optical differential spectroscopy and EPR, a parallel study of light-induced electron transfer between the primary (X1) and secondary (X2) quinone-like acceptors in the preparations of reaction centers (RC) isolated from bacterial chromatophore membranes with sodium dodecyl sulfate was carried out. The data from direct measurements of the rate constant temperature dependence for the interaction between light-reduced X1 and X2 (KX1X2) are in good agreement with the data calculated from the kinetic analysis of dark reduction of photooxidized bacteriochlorophyll RC on the acceptors X1 and X2 (KX1X2 = 2.10(-1)S at 20 degrees; Ea = 11,8 kcal.mol-1 within the temperature range of 20 degrees-- -20 degrees). This evidence proves the efficiency of the previously used approach /1, 2/ for the evaluation of the X1-X2 interaction. The method proposed was used for a kinetic analysis of a low-temperature electron transfer from X1 to X2 in RC isolated with lauryldimethylaminoxide (KX1X2 = 2,3.10(2) S-1 at 20 degrees; Ea = 5,5 kcal.mol-1 within the temperature range of 10 degrees-- --70 degrees).     

Preferential inhibition by ethidium bromide of photosynthetic apparatus formation in Rhodopseudomonas spheroides cells

Ethidium bromide (EB) inhibited growth of cells of the non-sulfurpurple photosynthetic bacterium Rhodopseudomonas spheroides.The inhibitory action of EB on the light-anaerobic culturedcells was stronger than on dark-aerobic cultured ones. EB alsosuppressed the induced synthesis of bacteriochlorophyll (Bchl)and carotenoids in the dark-aerobically grown cells incubatedwith gentle aeration under no-growth conditions, suggestingthat the target of the inhibitory action of EB on the photosyntheticgrowth of R. spheroides cells is chromatophore formation. EBdepressed the incorporation of ³H- or ¹⁴C-uracil into both RNAand DNA fractions from cells incubated with gentle aeration.In contrast, inhibition by EB of ³H-uracil incorporation intothe DNA fraction was not observed under vigorous aeration. Ourfindings seemed to favor the hypothesis described previously(10) that lowering the intracellular oxidation-reduction potentialmight bring about a unique synthesis or turnover of DNA responsiblefor chromatophore formation. (Received December 22, 1977; )     

Isolation and fractionation of the photosynthetic membranous organelles from Rhodopseudomonas spheroides

Molecular sieve chromatography and sucrose gradient centrifugation were used to prepare large quantities of purified chromatophores from Rhodopseudomonas spheroides. Electron micrographs of these chromatophores revealed that the final preparations were very homogeneous and free of non-chromatophore particulate material. As an additional check on purity, (14)C-l-phenylalanine-labeled aerobic cells, devoid of chromatophores, were mixed with unlabeled photosynthetic cells. The resulting preparation contained less than 1% of the radioactivity, originally located in non-chromatophore protein. The purified chromatophores were solubilized in 2-chloroethanol and separated into two fractions. Fraction P(1) contained 3 to 5% of the total chromatophore protein and could be resolved into 10 electrophoretic components. The second fraction, P(II), contained five electrophoretic components. One of these components had associated with it all of the pigment and phospholipid present in P(II). Preliminary immunochemical studies on these fractions are also reported.     

Light-induced absorbance change of carotenoid in chromatophores of photosynthetic bacterium, Rhodopseudomonas spheroides

The nature of the light-induced absorbance change of carotenoid,spheroidene, was investigated with the chromatophores of Rhodopseudomonasspheroides. The experimental results indicate that the changedoes not represent an oxidation-reduction reaction of the carotenoid,but is caused by a change in the state of the chromatophoresclosely related to the high energy state of the photophosphorylation.Since the change almost vanishes at liquid nitrogen temperature,it probably does not represent a primary photochemical reactionin the chromatophores. The values of the quantum yield for thechange of carotenoid were above unity ; 2.5 on an avera (Received November 20, 1969; )     

光合细菌Rhodopseudomonas产氢的影响因子实验研究

利用光合细菌Rhodopseudomonas 8株菌株研究初期活性、光照强度、C源种类、菌株差异对生物产氢的影响表明 ,不同菌株的C源利用性有较大差异 ,但对乳酸钠都有很好的利用性 .细菌的初期活性对产氢有一定程度的影响 ,稳定生长期的细菌比对数生长期的细菌产氢活性略高 .光照强度对产氢活性的影响明显 ,在光饱和度以下 ,光照强度大则产氢速率高 .不同菌种的产氢性能有效大差异 ,从上海地区有机污染环境中分离到的RhodopseudomonasB2 1 菌株在以乳酸钠 ( 50mmol·L- 1 )为C源、谷氨酸钠 ( 1 0mol·L- 1 )为N源 ,60 0 0Lx光照、30℃下 ,最大产氢速率达到 1 4.7ml·h- 1 ·g- 1 细胞干重 .