Isobutanol production from cellobiose in Escherichia coli

Converting lignocellulosics into biofuels remains a promising route for biofuel production. To facilitate strain development for specificity and productivity of cellulosic biofuel production, a user friendly Escherichia coli host was engineered to produce isobutanol, a drop-in biofuel candidate, from cellobiose. A beta-glucosidase was expressed extracellularly by either excretion into the media, or anchoring to the cell membrane. The excretion system allowed for E. coli to grow with cellobiose as a sole carbon source at rates comparable to those with glucose. The system was then combined with isobutanol production genes in three different configurations to determine whether gene arrangement affected isobutanol production. The most productive strain converted cellobiose to isobutanol in titers of 7.64?±?0.19 g/L with a productivity of 0.16 g/L/h. These results demonstrate that efficient cellobiose degradation and isobutanol production can be achieved by a single organism, and provide insight for optimization of strains for future use in a consolidated bioprocessing system for renewable production of isobutanol.     

Primary Infection of Barley by Erysiphe graminis f. sp. hordei in Relation to Leaf-Age Dependent Resistance and the Roles of the Epidermis and Mesophyll in this Resistance

Abstract The infection frequency of both compatible and incompatible races of Erysiphe graminis f. sp. hordei decreased gradually with increasing leaf age on undetached primary barley leaves. The length of secondary hyphae of the compatible race was approximately the same regardless of age, but secondary hyphae were slightly longer on younger seedlings than on older seedlings in the case of the incompatible race. Both the infection frequency and length of secondary hyphae of the two races weredistinctly different. On composite sections produced by exchanging the epidermal layers of young and relatively mature primary leaves, the infection frequency of the compatible race was higher on the epidermis of young leaves than on the epidermis of older, leaves, regardless of which mesophyll was under the epidermis. The epidermis appears to play a major role in age-dependent resistance, while the mesophyll may act disparately by providing a factor promotive to mildew infection in addition to supporting the resistance function of the epidermis.     

Inhibition of ATPase Activity in Pea Plasma Membranes In Situ by a Suppressor from a Pea Pathogen, Mycosphaerella pinodes

A pathogenic fungus of pea, Mycosphaerella pinodes, secretesa so-called "suppressor" in its pycnospore germination fluid.The suppressor blocks the defense responses and induces localsusceptibility (accessibility) in pea plants to agents thatare not pathogenic in pea. The suppressor nonspecifically inhibitsthe ATPase activity in plasma membranes prepared from pea, soybean,kidney bean, cowpea and barley plants. However, cytochemicalstudies by electron microscopy indicate that the suppressorspecifically inhibits the ATPase in pea cell membranes, butnot in those of four other plant species tested. That is, thespecificity of the suppressor appears at the cell and/or tissuelevel, but is not evident in vitro. Furthermore, the inhibitoryeffect of the suppressor is temporary because the ATPase activityrecovers 9 h after the treatment. A similar effect was observedafter inoculation with M. pinodes but not with a nonpathogenof pea, M. ligulicola. The role of the suppressor in host-parasitespecificity is discussed. (Received April 9, 1991; Accepted August 6, 1991)     

Inhibition of virus multiplication by immunoactive peptides

In the present study we show that peritoneal macrophages obtained from the mice treated with the immunoactive peptides inhibit the multiplication of Herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), but not that of vesicular stomatitis virus (VSV), and that the intraperitoneal administration of the peptides suppresses the infection with HSV-1 in mice.     

Stimulatory effect of calcium on photoactivation of the water oxidation system in dark-grown spruce chloroplasts

Treatment of platelets or red cells with small amounts of phospholipase C from Clostridium welchii enables both cells, prior to the onset of lysis, to stimulate prothrombin conversion by coagulation factor Xa and Va in the presence of calcium. Phospholipase C treatment of both cells also exposes significant amounts of phosphatidylserine at the outer surface. The level of phosphatidic acid formed from diglycerides produced by phospholipase C action, is similar to that formed in activated platelets upon triggering the phosphatidylinositol cycle. A possible involvement of this cycle to activate platelets to become more procoagulant is discussed.     

Effect of Streptococcus faecalis BIO-4R on intestinal flora of weanling piglets and calves.

The effect of oral administration of Streptococcus faecalis BIO-4R, an antibiotic-resistant lactic acid bacterium, on the intestinal flora of weanling piglets and cows reared on antibiotic-containing diet was investigated. Fourteen days after administration of the bacteria, the intestinal flora of the piglets was examined. Animals of the administered group had stabilized lactic flora such as bifidobacteria, streptococci, and lactobacilli, whereas most animals of control group had reduced lactic flora. On the other hand, abundant yeasts were detected from the cecum, colon, and feces of the control animals, but the levels were significantly lower in the animals given strain BIO-4R. The density of Salmonella in the intestine appeared to be reduced after the administration of strain BIO-4R. The number of BIO-4R cells was shown to be 10 times lower in the duodenum and jejunum than in the ileum, suggesting that strain BIO-4R might have grown transiently in the ileum. The similar trend toward stabilization of the lactic flora was also observed in cows after administration of BIO-4R. In addition, an antagonistic effect of the strain against yeasts and Salmonella was suggested. These findings indicate that the oral administration of strain BIO-4R is one of the useful methods whereby the potentially deleterious effect of antibiotics on the intestinal flora of farm animals may be minimized.     

Typing of Verotoxins by DNA Colony Hybridization with Poly- and Oligonucleotide Probes,a Bead-Enzyme-Linked Immunosorbent Assay,and Polymerase Chain Reaction

To identify the type of Verotoxins (VT) produced by Verocytotoxin-producing Escherichia coli (VTEC), a sensitive bead-enzyme-linked immunosorbent assay and polymerase chain reaction with common and specific primers to various VTs (VT1, VT2, VT2vha, VT2vhb, and VT2vp1) were developed. Together with colony hybridization tests with oligo- and polynucleotide probes, these methods were applied to VTEC isolates to type the VT produced. The toxin types of 26 of 37 strains were identified, but the reaction profiles in assays of the remaining 11 strains suggested the existence of new VT2 variants. The application of these identification procedures may be useful as a tool for clinical and epidemiological studies of VTEC infection.     

Photoactivation of the latent water-oxidizing complex in photosystem II membranes isolated from dark-grown spruce seedlings

Photosystem II membranes (D-PSII) were isolated from dark-grown spruce seedlings. All major PSII proteins except the 17- and 23-kDa extrinsic proteins were present in D-PSII. O₂ evolution and Mn content in D-PSII were negligible, while PSII-donor activity showed a value comparable to that of NH₂OH-treated PSII membranes (NH₂OH-L-PSII) from light-grown seedlings. Light incubation of D-PSII with 1 m M MnCl₂, 50 m M CaCl₂ and 100 μ M DCIP at pH 5.3 resulted in activation of the latent water-oxidizing complex. Accomplishment of photoactivation of PSII membranes from dark-grown spruce seedlings clearly indicates that only ligation of Mn²⁺ to the apo-water oxidizing complex is required for expression of O₂ evolution, and that protein synthesis is not involved in the photoactivation process. There was no essential difference between 'photoactivation' of naturally Mn-free PSII membranes and 'photoreactivation' of artificially Mn-depleted PSII membranes on kinetics, pH dependence, Mn²⁺-concentration dependence. However, kinetics and pH dependence of photoactivation were appreciably different in spruce PSII membranes and in PSII membranes of angiosperms such as wheat and spinach.