Large-scale production of GDP-fucose and Lewis X by bacterial coupling

A large-scale production system of GDP-fucose (GDP-Fuc) and fucosylated oligosaccharides was established by the combination of recombinant Escherichia coli cells overexpressing GDP-Fuc biosynthetic genes and Corynebacterium ammoniagenes cells. E. coli cells overexpressed the genes for glucokinase, phosphomannomutase, mannose-1-phosphate guanylyltransferase, GDP-mannose (GDP-Man) dehydratase, and GDP-4-keto-6-deoxy-mannose (GKDM) epimerase/reductase as well as phosphoglucomutase and phosphofructokinase. C. ammoniagenes contributed to the formation of GTP from GMP. GDP-Fuc accumulated to 29 mM (18.4 g l⁻¹) after a 22-h reaction starting with GMP and mannose through introducing the two-step reaction to overcome the inhibition of GDP-Fuc on GDP-Man dehydratase activity. When E. coli cells overexpressing the α1,3-fucosyltransferase gene of Helicobacter pylori were put into the GDP-Fuc production system, Lewis X [Galβ1–4(Fucα1–3)GlcNAc] was produced at an amount of 40 mM (21 g l⁻¹) for 30 h from GMP, mannose, and N-acetyl lactosamine. The production system through bacterial coupling can be applied to the industrial manufacture of fucosylated oligosaccharides. Journal of Industrial Microbiology & Biotechnology (2000) 25, 213–217. Received 01 May 2000/ Accepted in revised form 20 July 2000     

Early Tertiary fossil plants and paleoclimate of Lanzhou Basin

Fossil plants from the lower part of Xianshuihe Formation in the Lanzhou Basin, Gansu Province were studied. The flora contains 29 species, representing 20 genera and 12 families, which include Lauraceae ( Daphnogene ), Lardizabalaceae ( Akebia ), Berberidaceae ( Berberis ), Ulmaceae ( Planera, Ulmus, Zelkova ), Betulaceae ( Alnus, Carpinus ), Myricaceae( Myrica ), Salicaceae ( Populus, Salix), Myrsinaceae(Ardisia), Rosaceae ( Prunus, Sorbus, Sorbaria, Spiraea ), Leguminosae ( Gleditsia, Sophora), Anacardiaceae (Rhus), Caprifoliaceae(Viburnum). An analysis of the floristic elements and their foliar physiognomy shows that most members of the flora are deciduous broad-leaved trees or shrubs with a few evergreen shrubs. The most noteworthy species is Rhus turcomanica which was present in the Middle Eocene to Late Eocene of Central Asia (Kazakhstan, Turkmenistan). Generally, Rhus turcomanica occurred at the same beds as Palibinia, an extinct fossil plant whose presence indicates a subtropical dry climate. Another species, Sorbaria callicomifolia Kornilova was present from the Early Oligocene to Early Miocene of Central Asia (Kazakhstan and Turkmenistan). According to an analysis of spores and pollen, this flora contains over 20 species. It is predominated by the angiosperm pollen. There appeared Ephedripites and Nitrariadites which were important elements in the dry area. Ephedripites was found from the Upper Cretaceous to Early Tertiary. Nitrariadites occurred in the Late Miocene, whereas Rhus turcomanica and Sorbaria callicomifolia were both reported in the subtropical dry area from the Middle Eocene to Early Oligocene. The latest record of Rhus turcomanica is from the Middle Eocene to Early Oligocene of Central Asia. The presence of this element in the lower part of Xianshuihe Formation may indicate that itsage is the latest stage of the Early Oligocene.     

Large Scale Structural Analysis of cDNAs in the Model Legume, Lotus japonicus

Lotus japonicus possesses certain characteristics suited to molecular genetic and genomic analyses and has been adopted as a model species in the study of legume plants. To make a catalogue of genes expressed in L. japonicus and understand biological processes specific to legume plants, large scale EST analyses have been performed. To date, more than 26,000 EST sequences of L. japonicus have been deposited in the public databases. These sequences were developed by five laboratories using different organs. In this review, information obtained from two EST projects carried out in Japan is presented. Some 7137 non-redundant EST groups from young plants and 718 groups from flower buds were generated. A similarity search revealed that homologues of nodulin genes in other legume plants, as well as genes related to secondary metabolism, seed development and the reproductive process, were included in the EST collection, indicating the usefulness of the EST clones in the study of biological phenomena distinctive to legume plant species. Received 23 August 2000/ Accepted in revised form 22 September 2000     

Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110.

The complete nucleotide sequence of the genome of a symbiotic bacterium Bradyrhizobium japonicum USDA110 was determined. The genome of B. japonicum was a single circular chromosome 9,105,828 bp in length with an average GC content of 64.1%. No plasmid was detected. The chromosome comprises 8317 potential protein-coding genes, one set of rRNA genes and 50 tRNA genes. Fifty-two percent of the potential protein genes showed sequence similarity to genes of known function and 30% to hypothetical genes. The remaining 18% had no apparent similarity to reported genes. Thirty-four percent of the B. japonicum genes showed significant sequence similarity to those of both Mesorhizobium loti and Sinorhizobium meliloti, while 23% were unique to this species. A presumptive symbiosis island 681 kb in length, which includes a 410-kb symbiotic region previously reported by G?ttfert et al., was identified. Six hundred fifty-five putative protein-coding genes were assigned in this region, and the functions of 301 genes, including those related to symbiotic nitrogen fixation and DNA transmission, were deduced. A total of 167 genes for transposases/104 copies of insertion sequences were identified in the genome. It was remarkable that 100 out of 167 transposase genes are located in the presumptive symbiotic island. DNA segments of 4 to 97 kb inserted into tRNA genes were found at 14 locations in the genome, which generates partial duplication of the target tRNA genes. These observations suggest plasticity of the B. japonicum genome, which is probably due to complex genome rearrangements such as horizontal transfer and insertion of various DNA elements, and to homologous recombination.     

Imaging and measuring the rituximab-induced changes of mechanical properties in B-lymphoma cells using atomic force microscopy

The topography and mechanical properties of single B-lymphoma cells have been investigated by atomic force microscopy (AFM). With the assistance of microfabricated patterned pillars, the surface topography and ultrastructure of single living B-lymphoma cell were visualized by AFM. The apoptosis of B-lymphoma cells induced by rituximab alone was observed by acridine orange/ethidium bromide (AO/EB) double fluorescent staining. The rituximab-induced changes of mechanical properties in B-lymphoma cells were measured dynamically and the results showed that B-lymphoma cells became dramatically softer after incubation with rituximab. These results can improve our understanding of rituximab’effect and will facilitate the further investigation of the underlying mechanisms.     

烟青虫感染核型多角体病毒后围食膜的病变

昆虫的围食膜是衬在昆虫中肠内一种网状的结构,它可充作虫体抵御外来病原侵染的一道屏障。关于鳞翅目昆虫幼虫感染了昆虫病毒后围食膜的病变问题,国内外鲜有报道。尤锡镇和康慧娟(1985)曾以实验证明家蚕围食膜对核型多角体病毒有灭活作用,而且认为核型多角体病毒不能侵染和破坏围食膜。Derksen和Granados(1988)则证明染病幼虫的围食膜因不同杆状病毒(包括两种核型多角体病     

Toxicity of free proline revealed in an arabidopsis T-DNA-tagged mutant deficient in proline dehydrogenase

The toxicity of proline (Pro) to plant growth has raised questions despite its protective functions in response to environmental stresses. To evaluate Pro toxicity, we isolated an Arabidopsis T-DNA-tagged mutant, pdh, that had a defect in Pro dehydrogenase (AtProDH), which catalyzes the first step of Pro catabolism. The pdh mutant showed hypersensitivity to exogenous application of < or =10 mM L-Pro, at which wild-type plants grew normally. A dose-dependent increase in internal free Pro accumulation was observed in pdh plants during external Pro supply. These results do not just prove the toxicity of Pro, but also suggest that AtProDH is the only enzyme acting as a functional ProDH in ARABIDOPSIS: To further analyze the targets of Pro toxicity, we compared the expression of thousands of genes by pdh plants with that by wild-type plants by cDNA microarray analysis. Most genes were unaffected. Here we demonstrate Pro toxicity by using the pdh mutant and discuss a cause-and-effect action between an excess of free Pro and growth inhibition in ARABIDOPSIS.     

κ-卡拉胶琥珀酰基化衍生物的抗氧化活性研究

对κ-卡拉胶进行酸降解得到三种卡拉胶低聚糖,并进一步琥珀酰基化得到分子量分别为2720、4000和5960的κ-卡拉胶琥珀酰衍生物(A、B和C)。对产物进行FT-IR表征,并测得其琥珀酰基取代度(DS)分别为0.61、0.29和0.83。检测了三种κ-卡拉胶琥珀酰衍生物对超氧阴离子自由基O2.-、DPPH自由基、羟基自由基.OH以及过氧化氢的清除活性。结果表明:随着取代度的增加,其清除超氧阴离子自由基O2.-和DPPH自由基的能力增强;随着分子量的增加,其清除羟基自由基.OH和过氧化氢的能力增强。这可能与衍生物的羟基含量、取代基团的性质以及取代度等因素有关。     

ATP-independent strand transfer protein from murine spermatocytes, spermatids, and spermatozoa

A protein-catalyzing D-loop formation is present in murine spermatocytes, spermatids, and spermatozoa, but is not found in somatic tissue or in premeiotic cells of the germline. Unlike the Escherichia coli RecA protein and the meiotic rec protein (m-rec) previously described, D-loop formation by this protein (referred to as "mAi-rec") does not require ATP. The meiotic profile of mAi-rec activity is only partly similar to that of m-rec. Like m-rec, it rises steeply during early prophase and reaches a peak at pachytene. Unlike m-rec, its activity remains high during the postmeiotic phase of spermatid development and is prominent in immunochemically stained spermatozoa. A polyclonal antibody to E. coli RecA reacts with mAi-rec and inhibits its activity. No such reaction occurs with m-rec protein. The extent of sequence homology between E. coli RecA and murine mAi-rec is highly limited; none of the several monoclonal antibodies tested reacted with mAi-rec.