酸溶辅助超声波法提取类球红细菌类胡萝卜素条件优化

通过单因素和正交试验,对类球红细菌3757产类胡萝卜素的提取条件进行了研究。先采用超声波法、酸溶法、研磨法、冻融法、酸溶辅助超声波法和冻融辅助超声波法优化了从类球红细菌3757菌株中提取类胡萝卜素的方法,然后开展了酸溶辅助超声波法的单因素和正交实验,最后进行了重复性实验。结果表明,酸溶辅助超声波法是较优的提取方法,丙酮是较好的提取溶剂。最佳提取条件为料液比110、超声波处理总时间20 min、酸浓度3 mol/L、酸溶时间25 min、超声波振幅40%、超声工作/间隔时间2 min/1 min、酸溶温度27℃,实验重现较好。优化后类胡萝卜素的提取率较优化前提高了74.8%,为其产业化创造了条件。     

产氢红杆菌类胡萝卜素含量和组分分析

类胡萝卜素在调节光合细菌产氢中具有重要作用。采用丙酮-甲醇有机溶剂法和KOH甲醇皂化法对产氢红杆菌(Rhodobacter sp.)R7菌株类胡萝卜素进行了提取纯化,并进一步采用硅胶G薄板层析法对提取的类胡萝卜素进行了分离,并结合光谱法对分离的类胡萝卜素进行了定性和定量分析。结果表明,丙酮-甲醇(7∶2,V/V)提取3次可将色素提取完全;最佳提取时间为2h;超声波处理与否对提取率影响不明显;该工艺提取类胡萝卜素产率为2.81mg/g湿菌体。硅胶G薄板层析表明该菌株类胡萝卜素有4个主要组分:黄色、红色、浅红色和浅黄色,黄色和红色为主要成分,光谱学数据显示黄色组分为球形烯,红色组分为螺菌黄质系类胡萝卜素,表明产氢红杆菌类胡萝卜素代谢途径独特。     

产氢红杆菌类胡萝卜素含量和组分分析

类胡萝卜素在调节光合细菌产氢中具有重要作用。采用丙酮－甲醇有机溶剂法和KOH甲醇皂化法对产氢红杆菌（Rhodobacter sp.）R7菌株类胡萝卜素进行了提取纯化,并进一步采用硅胶G薄板层析法对提取的类胡萝卜素进行了分离,并结合光谱法对分离的类胡萝卜素进行了定性和定量分析。结果表明,丙酮－甲醇（7∶2,V/V）提取3次可将色素提取完全;最佳提取时间为2h;超声波处理与否对提取率影响不明显;该工艺提取类胡萝卜素产率为2.81mg/g湿菌体。硅胶G薄板层析表明该菌株类胡萝卜素有4个主要组分：黄色、红色、浅红色和浅黄色,黄色和红色为主要成分,光谱学数据显示黄色组分为球形烯,红色组分为螺菌黄质系类胡萝卜素,表明产氢红杆菌类胡萝卜素代谢途径独特。     

提取方法对库拉索芦荟多糖的抗氧化活性影响研究

文章探讨了水提法和超声波辅助复合酶法对芦荟多糖抗氧化活性的影响。以多糖提取率和多糖含量为评价指标，分别得到水提法和超声波辅助复合酶法的提取产物，采用体外抗氧化活性体系评价2种提取方法对芦荟多糖抗氧化活性的影响。抗氧化活性评价结果显示，超声波辅助复合酶法对于多糖提取物的羟基自由基清除率和DPPH自由基清除率均高于水提法。     

红酵母生物合成类胡萝卜素研究进展

类胡萝卜素(carotenoid)是一类呈黄色、橙红色或红色的多烯类化合物,具有预防血管硬化和抑制肿瘤发生,增加宿主免疫力,抗氧化能力和抑制自由基等功能.目前类胡萝卜素已被FAO和WHO等国际组织定为A类营养色素,并在50多个国家获准为营养、色素双重功用的食品添加剂,被广泛应用于保健食品及医药和化妆品工业[1].生产类胡萝卜素可采用提取法、化学合成法和微生物发酵法,其中最有发展前景的是微生物发酵法.目前国内外微生物发酵技术生产天然类胡萝卜素的研究,主要集中在三孢布腊拉霉菌和红酵母等菌种上.利用红酵母生产类胡萝卜素具有营养要求简单,培养周期短,可综合利用等优点,具有很好的应用价值和开发前景.本文主要对红酵母生物合成类胡萝卜素的菌种选育、培养条件及类胡萝卜素的提取检测方法进行了综述.     

响应面优化食用菌多酚的提取及抗氧化活性评价

目的：优化食用菌多酚的超声波法提取工艺,对比不同食用菌多酚的抗氧化活性。方法：以多酚提取率为指标,采用超声波法,通过单因素试验及响应面分析,确定超声波法的最佳工艺流程。采用Folin-Ciocalteu比色法检测多种食用菌的多酚提取率,并以对DPPH·清除能力评价食用菌多酚的抗氧化活性。结果：响应面试验确定超声波法的最佳提取条件为乙醇浓度为85%、超声时间为38 min、液料比为44∶1mL/g,多酚提取率可达5.258 mg/g,与模型理论预测值（5.236 mg/g）的相对误差仅为1.28%,表明该工艺稳定、可靠。食用菌多酚提取率与抗氧化能力检测结果表明,超声波法得到的食用菌多酚都具有抗氧化活性,且抗氧化能力与多酚提取率之间的相关性显著,其中黄牛肝菌单位毫克多酚的DPPH·清除能力最强。结论：通过响应面法优化超声波法提取食用菌多酚的工艺条件,保持了食用菌多酚的抗氧化活性,是一种可行、实用和高效的食用菌多酚提取方法。     

酶法辅助超声波法提取类球红细菌辅酶Q10条件优化

采用单因素和正交实验优化了酶法辅助超声波法提取类球红细菌辅酶Q10的提取条件。结果表明,较优的辅酶Q10提取条件:超声波总时间14 min,超声波振幅30%,超声波工作/间歇时间1 min/1 min,料液比115,溶菌酶添加量300μL,酶解pH 7.2,酶解温度37℃,酶解时间90 min。优化后辅酶Q10的提取率比优化前提高了91.9%。     

优美红游动菌类胡萝卜素的提取条件研究

研究了优美红游动菌(Rhodoplanes elegans)的生长以及类胡萝卜素的产生,通过比较4种广泛采用的细胞破碎方法,对优美红游动菌(Rhodoplanes elegans)类胡萝卜素的提取条件进行了研究,结果表明:类胡萝卜素的产生与菌种的生长曲线相符合,菌种活化后培养45h,类胡萝卜素的含量趋于稳定;取此时培养液,采用超声波法破碎细胞(破碎功率640W,时间10min),类胡萝卜素提取效果最好,初提液类胡萝卜素的提取率约为7.62mg·(g干菌体)^-1;酸热法对色素的破坏严重。这为进一步开发利用天然色素资源提供了一定的理论依据。     

桦褐孔菌多糖的提取工艺优化及抗氧化活性评价

目的：优化桦褐孔菌多糖的超声波法提取工艺,对比不同产地桦褐孔菌多糖的抗氧化活性。方法：采用正交试验法确定超声波法的最佳工艺流程,检测不同产地桦褐孔菌的多糖提取率和抗氧化活性。结果：超声波法的最佳提取条件为超声时间25min、料液比1∶40、超声温度50℃、提取3次,在此条件下多糖提取率高达8.61 g/100g,与水提醇沉法相比提高了17.3%,耗时缩短了3/4,大大提高了提取效率。抗氧化能力检测结果证明超声波法得到的桦褐孔菌多糖都具有抗氧化活性,并且抗氧化能力与多糖提取率之间的相关性极其显著。结论：通过优化提取条件,超声波法明显提高了多糖提取率,保持了桦褐孔菌多糖的抗氧化活性,是一种可行、实用和高效的真菌多糖提取方法。     

含硒类球红细菌的研究

为了确定类球红细菌转硒培养的最佳条件 ,研究了无机硒的加入浓度、时间以及分批补料培养对菌体生长和转硒效率的影响。实验表明 ,无机硒的浓度低于 1× 10 -5mol/L时 ,对类球红细菌的生长基本没有影响 ,并能将6 3.9%的无机硒转化为有机硒。转硒的最佳时间是在接种后 12h左右 ,此时转硒效率最高。实验还表明 ,分批补料培养可以提高菌体浓度 ,可使转硒效率和绝对量增加。体内试验表明 ,用 5mL/kgbw和 10mL/kgbw剂量的含硒类球红细菌灌养小鼠 ,可以使其全血GSH Px酶活性提高 2 0 .9%和 2 5 .5 % ,使其血清丙二醛 (MDA)含量降低2 1.0 %和 2 3.2 %。     

类球红细菌应用进展

类球红细菌(Rhodobacter Sphaeroides)属光合细菌,是目前研究最深入的光合微生物之一,具有多种代谢方式。不仅能够产生类胡萝卜素、辅酶Q10、超氧化物歧化酶、5-氨基乙酰丙酸和氢气,而且能够降解农药残留、有机废水和多环芳烃等有毒有害物质,应用领域十分广泛。本文综述了类球红细菌在食品、医药、农业、环境、氢气生产等领域中的应用进展,并对类球红细菌的应用前景进行展望。     

Differences in the control of bacteriochlorophyll formation by light and oxygen

Control of bacteriochlorophyll formation was studied with continuous cultures of Rhodospirillum rubrum, Rhodopseudomonas sphaeroides, and Rhodopseudomonas capsulata. Oxygen controlled specific bacteriochlorophyll contents of the three species in a hyperbolical fashion irrespective of the presence of light. In Rps. sphaeroides, this applied to oxygen concentrations above 16% air saturation of the medium while at lower oxygen concentrations control followed a kinetics with negative cooperativity. Cell protein formation of R. rubrum and Rsp. sphaeroides was independent of oxygen concentrations while protein formation of Rps. capsulata increased at lower concentrations. Light controlled bacteriochlorphyll contents of R. rubrum and Rps. sphaeroides in a sigmoidal fashion. When growing at a constant low oxygen concentration cell protein formation increased with light energy flux in Rps. sphaeroides but remained unaffected in R. rubrum. Protein formation of R. rubrum increased with light energy flux only under anaerobic conditions. Two factor analyses were performed with R. rubrum and Rps. sphaeroides to study the combined effects of light and oxygen on bacteriochlorophyll formation. The results showed that both factors act independent of each other.Abbreviations ALA 5-aminolevulinic acid - R Rhodospirillum - Rsp. Rhodopseudomonas     

Heterologous synthesis and assembly of functional LHII antenna complexes from <Emphasis Type="Italic">Rhodovulum sulfidophilum</Emphasis> in <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis> mutant

The light harvesting complexes, including LHII and LHI, are the important components of photosynthetic apparatus. Rhodovulum (Rdv.) sulfidophilum and Rhodobacter (R.) sphaeroides belong to two genera of photosynthetic bacteria, and they are very different in some physiological characteristics and light harvesting complexes structure. The LHII structural genes (pucBsAs) from Rdv. sulfidophilum and the LHI structural genes (pufBA) from R. sphaeroides were amplified, and cloned into an expression vector controlled by puc promoter from R. sphaeroides, which was then introduced into LHI and LHII-minus R. sphaeroides mutants; the transconjugant strains synthesized heterologous LHII and native LHI complexes, which played normal roles in R. sphaeroides. The Rdv. sulfidophilum LHII complex from pucBsAs had near-infrared absorption bands at ~801–853 nm in R. sphaeroides, and was able to transfer energy efficiently to the native LHI complex. The results show that the pucBsAs genes from Rdv. sulfidophilum could be expressed in R. sphaeroides, and the functional foreign LHII and native LHI were assembled into the membrane of R. sphaeroides.     

Mutants of Rhodopseudomonas sphaeroides useful in genetic analysis

Two kinds of mutants of Rhodopseudomonas sphaeroides that should be useful in extending genetic analysis of this organism have been isolated. One is deficient in recombination and has been used to isolate derivatives of the plasmid R 68.45 which incorporate chromosomal genes of R. sphaeroides. The other is apparently defective in a DNA restriction enzyme; transfer of plasmid borne chromosomal genes of R. sphaeroides from Escherichia coli back to R. sphaeroides is greatly enhanced in these mutants.In memory of R. Y. Stanier     

Expression of four <Emphasis Type="Italic">pha</Emphasis> genes involved in poly-β-hydroxybutyrate production and accumulation in <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis> FJ1

A cluster of genes encoding polyhydroxybutyrate (PHB) depolymerase (phaZ), PHB synthase (phaC), phasin (phaP), and the regulator protein (phaR) was previously identified in Rhodobacter sphaeroides FJ1 (R. sphaeroides FJ1). In this study, we investigated the role of the PhaR protein on the expression of the pha genes. Immunoblot analysis revealed that the expressions of phaP, phaZ and phaR genes in wild-type cells of R. sphaeroides FJ1 are repressed during the active growth phase, with the exception of phaC. A phaR deletion mutant of R. sphaeroides FJ1 was constructed, and the basal level of phaP and phaZ expression in this mutant was markedly increased. Electrophoretic mobility shift assays demonstrated that PhaR binds to the promoter region of phaP as well as those of phaR and phaZ. These results suggest that the PhaR protein is a repressor of phaP, phaR, and phaZ genes in R. sphaeroides FJ1.     

Purification and properties of the adenylate kinases from Rhodopseudomonas palustris,Rhodopseudomonas sphaeroides and Rhodopseudomonas rubrum

The adenylate kinases (EC 2.7.4.3) from photosynthetically grown Rhodopseudomonas palustris, Rhodopseudomonas sphaeroides and Rhodospirillum rubrum were purified to homogeneity by the same procedure. The purified enzymes showed optimal rates of activity with MgCl₂ at 25° C and pH 8.0. They were found to be heat labile and were characterized by pI-values of 4.5. Apparent molecular weights of 33 500 for R. palustris, 34 400 for R. sphaeroides and 32 100 for R. rubrum were determined by high performance liquid chromatography. No separation into subunits was observed by use of sodium dodecylsulfate polyacrylamide gel electrophoresis. The apparent K _m -values for ADP corresponded to 0.26 mM for R. palustris, 0.27 mM for R. sphaeroides and 0.24 mM for R. rubrum. ADP in excess had a strong inhibitory effect. Competitive product inhibition was found for AMP, with K _i-values of 0.017 mM for R. palustris, 0.018 mM for R. sphaeroides and 0.014 mM for R. rubrum. A competitive inhibitor likewise was P¹,P⁵-di(adenosine-5)pentaphosphate with K _i-values of 0.020 M for R. palustris and R. sphaeroides, and 0.017 M for R. rubrum. Sulfhydryl-reacting reagents like p-chloromercuribenzoate and iodoacetic acid were found to be non-inhibitory. All measurements of adenylate kinase activity were carried out with the stabilized and most sensitive luciferin-luciferase system.     

Improvement of lipid profile and antioxidant of hypercholesterolemic albino rats by polysaccharides extracted from the green alga Ulva lactuca Linnaeus

Sulfated polysaccharides from Ulva lactuca were extracted in hot water and precipitated by ethanol then orally gavaged to rats fed on a hypercholesterolemic diet for 21 days to evaluate the antihypercholesterolemic and antioxidant actions. Atorvastatine Ca (Lipitor) was used as a reference drug. The intragastric administration of U. lactuca extract to hypercholesterolemic rats caused significant decrease of serum total lipids, triglycerides, total cholesterol, LDL-cholesterol and vLDL-cholesterol levels. Whereas, HDL-cholesterol concentration was markedly increased by 180%. Aqueous extract showed a significant ameliorative action on elevated atherogenic index, creatine kinase and lactate dehydrogenase activities of hypercholesterolemic group. Furthermore, serum activities of transaminases and alkaline phosphatase were also improved. High fat diet intake caused a highly significantly elevated serum urea, creatinine concentration. These effects were reversed by oral administration of U. lactuca extract. Sulfates polysaccharides extract of U. lactuca ameliorate hepatic enzymatic (catalase, glutathione peroxidase and superoxide dismutase), non-enzymatic (reduced glutathione & total thiol) antioxidant defenses and thiobarbituric acid reactive substances. In conclusion, the tested U. lactuca polysaccharides extract has potent hypocholesterolemic and antioxidant effects in experimentally-induced hypercholesterolemic animal model.     

Molecular cloning, sequence and regulation of expression of the recA gene of the phototrophic bacterium Rhodobacter sphaeroides

The recA gene of Rhodobacter sphaeroides 2.4.1 has been isolated by complementation of a UV-sensitive RecA^– mutant of Pseudomonas aeruginosa. Its complete nucleotide sequence consists of 1032 bp, encoding a polypeptide of 343 amino acids. The deduced amino acid sequence displayed highest identity to the RecA proteins from Rhizobium mehloti, Rhizobium phaseoli, and Agrobacterium tumefaciens. An Escherichia coli-like SOS consensus region, which functions as a binding site for the LexA repressor molecule was not present in the 215 by upstream region of the R. sphaeroides recA gene. Nevertheless, by using a recA-lacZ fusion, we have shown that expression of the recA gene of R. sphaeroides is inducible by DNA damage. A recA-defective strain of R. sphaeroides was obtained by replacement of the active recA gene by a gene copy inactived in vitro. The resulting recA mutant exhibited increased sensitivity to UV irradiation, and was impaired in its ability to perform homologous recombination as well as to trigger DNA damage-mediated expression. This is the first recA gene from a Gram-negative bacterium that lacks an E. coli-like SOS box but whose expression has been shown to be DNA damage-inducible and auto-regulated.     

Activity and characterization of mixed organic compounds extracted from Rhodobacter sphaeroides as alternative materials to serum for mammalian cell growth

Photosynthetic microorganisms produce relatively large amounts of physiologically active materials which stimulate the physiological activity of other organisms. In this study, mammalian HeLa cells were cultured in different culture media which were Dulbecco's modified Eagle medium (DMEM) with newborn calf serum (NCS), and DMEM including different types of physiologically activating compounds (PACs) extracted from Rhodobacter sphaeroides grown under various culture conditions. R. sphaeroides was grown under the following five different culture conditions: anaerobically in the light, anaerobically in the dark and treated with dimethyl sulfoxide, aerobically in the dark for 48 h, in the light for 48 h, and in the light for 24 h and changed after previous culturing in the dark for 24 h. The growth of HeLa cell was measured by cell counting using a hemocytometer, and the fluorescent intensities of cellular lysosomes were measured to check the level of cellular stress caused by adding PACs. The growth of HeLa cells cultured in DMEM with PACs extracted from R. sphaeroides aerobically grown under dark conditions was enhanced compared to that of cells grown with NCS. We also found that a high concentration of pigments such as bacteriochlorophylls and carotenoids and a high concentration of arginine produced by R. sphaeroides aerobically grown in the dark were implicated in increased growth of the HeLa cells. Therefore, our results suggest that PACs extracted from R. sphaeroides aerobically cultured in dark conditions can enhance the physiological activity of mammalian cells and serve as nontoxic and bioavailable materials.     

DNA-DNA hybridization of Rhodopseudomonas capsulata,Rhodopseudomonas sphaeroides and Rhodopseudomonas sulfidophila strains

The genetic relatedness of 21 Rhodopseudomonas strains has been studied by means of DNA-DNA hybridization. All strains included in the study belonged to the subgroup of the genus Rhodopseudomonas which is characterized by a short-rod to coccus morphology, a vesicular intracytoplasmic membrane system and carotenoids of the spheroidene group. Mol percentages guanine + cytosine ranged from 64 to 73, most strains having values between 68 and 72. With few exceptions, the hybridization data obtained were in agreement with the subdivision in three (or possibly four) species on the basis of classical taxonomy. Strain SCJ, formerly considered to be a somewhat atypical R. capsulata strain, is most probably a R. sphaeroides strain and two out of seven strains that were received as R. sulfidophila did not fit in this species on the basis of the hybridization data. The results also showed that two undesignated strains that were previously thought to be related to R. capsulata (Hansen et al. 1975) cannot be assigned to this species and may be representatives of another species. The seven strains that required approximately 2.5% NaCl in the medium and that had been designated R. sulfidophila were found to synthesize far higher levels of bacteriochlorophyll during fully aerobic growth in the dark than the purple bacteria studied thus far.Abbreviations GC guanosine + cytosine - SSC standard saline citrate buffer