Biotransformation of l-ornithine from l-arginine using whole-cell recombinant arginase

A recombinant arginase was generated for a whole-cell biotransformation system to convert l-arginine to l-ornithine in Escherichia coli. The gene ARG1 coding arginase from Bos taurus liver was synthesized and expressed in E. coli BL21 (DE3) via pETDuet-1. The recombinant arginase was used to catalyze l-arginine to l-ornithine and urea. The reaction was optimal at pH 9.5 and 37 °C. Manganese (10^?5 M) and Emulsifier OP-10 [0.033 % (v/v)] could promote arginase activity. In a scale up study, l-arginine conversion rate reached 98 % with a final concentration of 111.52 g l-ornithine/l.     

Mortality factors of satin moth,Leucoma salicis [Lep.: Lymantriidae], in aspen forests in Maine

Natural control agents of the satin moth,Leucoma salicis (L.) were examined in 2Populus grandidentata Michaux stands. Highest mortality occurred in overwintering larvae, with the causal agents 2 fungi,Paecilomyces sp. andHirsutella gigantea Petch, a factor causing death symptomatic of a pathogen, the parasiteEupteromalis hemipterus (Walker) and death from unknown causes. Mortality fromPaecilomyces sp. andE. hemipterus was reduced where overwintering sites were covered with burlap cloth. Parasites recovered from larvae and pupae included the braconidsApanteles melanoscelus (Ratzeburg) andMeteorus versicolor (Wesmeal), the ichneumonidCoccygomimus pedalis (Cresson), the tachinidsCompsilura concinnata (Meigen),Carcelia laxifrons Villeneuve,Tachinomyia variata Curran, and the sarcophagidsSarcophaga aldrichi Parker andAgria housei Shewell. Larval and pupal predators included the carabidCalosoma frigidum Kirby, pentatomids, and birds, particularly black-billed cuckoos,Coccyzus erythrophthaimus (Wilson). Eggs were parasitized by the scelionidTelenomus prob.californicus Ashmead and the trichogrammidTrichogramma minutum (Riley). Predators of adult satin moths included the hermit thrush,Hylocichla guttata (Pallas), and pentatomid bugs.     

The effect of prey density on the functional and numerical reponses of three species of predatory mites

Functional and numerical responses of the predators:Phytoseiulus persimilis Athias-Henriot,Metaseiulus occidentalis (Nesbitt), andAmblyseius chilenensis (Dosse) [Acarina, Phytoseiidae] were observed at prey (Tetranychus urticae Koch [Acarina, Tetranychidae]) densities up to 300 prey/6.45 cm². Neither functional nor numerical response curves revealed any prey-predator interference effects, i.e.: the dome-shaped response curves (Holling, 1961), did not occur.     

Characterization of a recombinant l-fucose isomerase from Caldicellulosiruptor saccharolyticus that isomerizes l-fucose, d-arabinose, d-altrose, and l-galactose

A recombinant l-fucose isomerase from Caldicellulosiruptor saccharolyticus was purified as a single 68 kDa band with an activity of 76 U mg^?1. The molecular mass of the native enzyme was 204 kDa as a trimer. The maximum activity for l-fucose isomerization was at pH 7 and 75°C in the presence of 1 mM Mn²⁺. Its half-life at 70°C was 6.1 h. For aldose substrates, the enzyme displayed activity in decreasing order for l-fucose, with a k _cat of 11,910 min^?1 and a K _m of 140 mM, d-arabinose, d-altrose, and l-galactose. These aldoses were converted to the ketoses l-fuculose, d-ribulose, d-psicose, and l-tagatose, respectively, with 24, 24, 85, 55% conversion yields after 3 h.     

Metabolic engineering Corynebacterium glutamicum for the l-lysine production by increasing the flux into l-lysine biosynthetic pathway

The experiments presented here were based on the conclusions of our previous results. In order to avoid introduction of expression plasmid and to balance the NADH/NAD ratio, the NADH biosynthetic enzyme, i.e., NAD-dependent glyceraldehyde-3-phosphate dehydrogenase (GADPH), was replaced by NADP-dependent GADPH, which was used to biosynthesize NADPH rather than NADH. The results indicated that the NADH/NAD ratio significantly decreased, and glucose consumption and l-lysine production drastically improved. Moreover, increasing the flux through l-lysine biosynthetic pathway and disruption of ilvN and hom, which involve in the branched amino acid and l-methionine biosynthesis, further improved l-lysine production by Corynebacterium glutamicum. Compared to the original strain C. glutamicum Lys5, the l-lysine production and glucose conversion efficiency (α) were enhanced to 81.0 ± 6.59 mM and 36.45 % by the resulting strain C. glutamicum Lys5-8 in shake flask. In addition, the by-products (i.e., l-threonine, l-methionine and l-valine) were significantly decreased as results of genetic modification in homoserine dehydrogenase (HSD) and acetohydroxyacid synthase (AHAS). In fed-batch fermentation, C. glutamicum Lys5-8 began to produce l-lysine at post-exponential growth phase and continuously increased over 36 h to a final titer of 896 ± 33.41 mM. The l-lysine productivity was 2.73 g l^?1 h^?1 and the α was 47.06 % after 48 h. However, the attenuation of MurE was not beneficial to increase the l-lysine production because of decreasing the cell growth. Based on the above-mentioned results, we get the following conclusions: cofactor NADPH, precursor, the flux through l-lysine biosynthetic pathway and DCW are beneficial to improve l-lysine production in C. glutamicum.     

Observation of algal colonization on Acropora aspera killed by Acanthaster planci

Algae started colonizing branches of the coral Acropora aspera (Dana) killed by the sea star Acanthaster planci (Linnaeus) within less than 24 hours. Two blue-greens ((Microcoleus lyngbyaceus (Crouan) Ag. and Hormothamnion solutum B. & F.)) dominated the early community but became less abundant than a brown ((Giffordia indica (Sonder) Papenfuss & Chihara)) after 26 days.     

Chromosome counts and chromosome morphology of some selected species

In the years 1976–1981 we studied chromosome counts and karyotypic formulae of the following 29 species of plants from 41 localities (of these 6 from Bohemia, 32 from Moravia, 3 from Slovakia):Batrachium baudotii (Godron) F. W. Schultz,Chenopodium rubrum L.,C. polyspermum L.,C. murale L.,C. ficifolium Sm.,C. opulifolium Schrader ex DC. inLam. et DC.,C. strictum Roth [subsp.strictum, subsp.glaucophyllum (Aellen)Aellen inJust etAellen, subsp.striatiforme Uotila],Arenaria grandiflora L.,Illecebrum verticillatum L.,Spergula morisonii Boreau inDuchartre,Spergularia marginata (DC. inLam. et DC.)Kittel S. marina (L.)Griseb.,S. rubra (L.) J. etC. Presl,Silene conica L.,Sisymbrium loeselii L.,S. volgense Bieb. exE. Fourn.,S. orientale L. [subsp. orientale, subsp.macroloma (A. Pomel)Dvo?ák],S. officinale (L.)Scop.,Descurainia sophia (L.)Webb exPrantl inEngler etPrantl,Nasturtium officinale R. Br. inAiton,Barbarea arcuata (Opiz inPresl J. et C.)Reichenb.,Lunaria annua L.,Soldanella montana Willd.,S. carpatica Vierh. inUrban etGraebner,Lotus tenuis Waldst. etKit. exWilld.,L. uliginosus Schkuhr,Trigonella monspeliaca L.,Geranium sibiricum L.,Lactuca tatarica (L.)C. A. Meyer.     

Impact of aerially appliedBacillus thuringiensis and carbaryl on gypsy moth [Lep.: Lymantriidae] and adult parasites

In 1973, Dipel^® (Bacillus thuringiensis Berliner) and Sevin^® 4 oil (carbaryl) were applied aerially on a dense population of gypsy moths,Lymantria dispar (L.), to evaluate the effect of these biological and chemical insecticides on gypsy moth larvae and adult parasites. Both insecticides provided excellent protection of foliage (29 % average defoliation) and gypsy moth population reduction (99 %). Significantly fewerBrachymeria intermedia (Nees), andApanteles melanoscelus (Ratzeburg),Parasetigena silvestris (Robineau-Desvoidy) andCompsilura concinnata (Meigen) adult parasites were captured in the blocks treated with Sevin 4 oil or Dipel than in the control block during the treatment year and in 1974, respectively.     

Cytotaxonomic study ofTragopogon L. in czechoslovakia

Karyotypes ofTragopogon orientalis L. subsp.orientalis, T orientalis L. subsp.leiocarpos (Sauter)Trnka,T. pratensis L.,T. minor Miller,T. dubius Scop. subsp.dubius andT. dubius Scop. subsp.major (Jacq.)Vollmann were studied. The occurrence in Slovakia ofT. pratensis was karyologically proved.     

Parasitism ofPhthorimaea operculella [Lep.: Gelechiidae] larvae in Queensland

Parasitism ofPhthorimaea operculella (Zeller) larvae on potato foliage was examined intensively in the Lockyer Valley, south-eastern Queensland from 1975–78 and extensively from other hosts and areas of Queensland. The % parasitism in the Lockyer Valley exceeded 50% in 29 of the 36 collections. The parasite species were dominated by the introducedCopidosoma desantisi Annecke & Mynhardt (Encyrtidae) andOrgilus lepidus Muesebeck (Braconidae) which together accounted for 92.6% of parasite numbers recorded. In more northern areas of Queensland, the dominant parasite species found was the introducedApanteles subandinus Blanchard (Braconidae). Other hymenopterous parasites found wereMicrochelonus curvimaculatus Cameron (Braconidae), Elasmus funereus Riek (Elasmidae) andTemelucha minuta (Morley) (Ichneumonidae).     

The kinetic analysis of the substrate specificity of motif 5 in a HAD hydrolase-type phosphosugar phosphatase of Arabidopsis thaliana

The Arabidopsis thaliana gene AtSgpp (locus tag At2g38740), encodes a protein whose sequence motifs and expected structure reveal that it belongs to the HAD hydrolases subfamily I, with the C1-type cap domain (Caparrós-Martín et al. in Planta 237:943–954, 2013). In the presence of Mg²⁺ ions, the enzyme has a phosphatase activity over a wide range of phosphosugar substrates. AtSgpp promiscuity is preferentially detectable on d-ribose-5-phosphate, 2-deoxy-d-ribose-5-phosphate, 2-deoxy-d-glucose-6-phosphate, d-mannose-6-phosphate, d-fructose-1-phosphate, d-glucose-6-phosphate, dl-glycerol-3-phosphate, and d-fructose-6-phosphate. Site-directed mutagenesis analysis of the putative signature sequence motif-5 (IAGKH), which defines its specific chemistry, brings to light the active-site residues Ala-69 and His-72. Mutation A69M, changes the pH dependence of AtSgpp catalysis, and mutant protein AtSgpp-H72K was inactive in phosphomonoester dephosphorylation. It was also observed that substitutions I68M and K71R slightly affect the substrate specificity, while the replacement of the entire motif for that of homologous dl-glycerol-3-phosphatase AtGpp (MMGRK) does not switch AtSgpp activity to the specific targeting for dl-glycerol-3-phosphate.     

Mechanical transmission of beet yellows virus toChenopodium quinoa Willd. andChenopodium foliosum (Moench) Asch.

Beet yellows virus (BYV) was mechanically transmitted by sap from sugar-beet plants, infected with BYV, to the plants ofChenopodium quinoa Willd. and ofChenopodium foliosum (Moench) Asch. Mechanical transmission of BYV to the plants ofTetragonia expansa Murr. failed. Infectious material was homogenized in phosphate buffer with veronal and EDTA, pH 7–8. Experimental plants were darkened three days before infection and kept at a temperature of 5°C. Plants ofC. quinoa Willd. were decapitated. Back transmissions fromC. quinoa Willd. andC. foliosum (Moench) Asch. infected with BYV, to sugar-beet plants were carried out by the aveidMyzus persicae Sulz. These transmissions were positive. Filamentous particles of BYV, of an average length 1275 nm, were found in plants ofC. quinoa Willd. andC. Foliosum (Moench) Asch., infected with BYV.     

Overexpression of d-psicose 3-epimerase from Ruminococcus sp. in Escherichia coli and its potential application in d-psicose production

The d-psicose 3-epimerase (DPE) gene from Ruminococcus sp. was cloned and overexpressed in Escherichia coli. The recombinant protein was purified and characterized. It was optimally active at pH 7.5–8.0 and 60?°C. Activity was not dependent on the presence of metal ions; however, it became more thermostable with added Mn²⁺. The K _m of the enzyme for d-psicose (48?mM) was lower than that for d-tagatose (230?mM), suggesting that d-psicose is the optimum substrate. More importantly, the thermostability of the novel DPE from Ruminococcus is the strongest among all of the d-psicose and d-tagatose 3-epimerases and may be suitable for the industrial production of d-psicose from fructose.     

Survey and distribution ofCybocephalidae [Coleoptera] in Israel

Nine species ofCybocephalus were found in a survey conducted throughout Israel:C. micans (Reitter),C. nigriceps nigriceps (J. Sahiberg),C. nigriceps sinister Endrödy-Younga,C. aegyptiacus Endrödy-Younga,C. mediterraneus Endrödy-Younga,C. pullus Endrödy-Younga,C. fodori minor Endrödy-Younga,C. membranaceus (Reitter) andCybocephalus sp. n.C. micans andC.n. nigriceps are the most prevalent: the former in the Coastal Plain on citrus and the latter in the Arava region on date palms. In most cases the population density of the predator appeared to be related to that of the prey. Armoured scale insects(Homoptera: Diaspididae) were the main prey ofCybocephalus.     

Backbone and side-chain 1H, 15N and 13C assignment of apo- and imipenem-acylated l,d-transpeptidase from Bacillus subtilis

The d,d-transpeptidase activity of Penicillin Binding Proteins (PBPs) is essential to maintain cell wall integrity. PBPs catalyze the final step of the peptidoglycan synthesis by forming 4 → 3 cross-links between two peptide stems. Recently, a novel β-lactam resistance mechanism involving l,d-transpeptidases has been identified in Enterococcus faecium and Mycobacterium tuberculosis. In this resistance pathway, the classical 4 → 3 cross-links are replaced by 3 → 3 cross-links, whose formation are catalyzed by the l,d-transpeptidases. To date, only one class of the entire β-lactam family, the carbapenems, is able to inhibit the l,d-transpeptidase activity. Nevertheless, the specificity of this inactivation is still not understood. Hence, the study of this new transpeptidase family is of considerable interest in order to understand the mechanism of the l,d-transpeptidases inhibition by carbapenems. In this context, we present herein the backbone and side-chain ¹H, ¹⁵N and ¹³C NMR assignment of the l,d-transpeptidase from Bacillus subtilis (Ldt_Bs) in the apo and in the acylated form with a carbapenem, the imipenem.     

The parasitoid complex of three noctuids [Lep.] in a northern Florida cropping system: Seasonal occurrence,parasitization, alternate hosts,and influence of host-habitat

Parasitoids of the cabbage looper,Trichoplusia ni (Hübner), the soybean looper,Pseudoplusia includens (Walker), and the tobacco budworm,Heliothis virescens (F.) were characterized in a 3.2-ha model of a north Florida (U.S.A.) cropping system (including tobacco (Nicotiana tabacum L.), soybeans (Glycine max (L.)Merr. and 18 other crops) not treated with chemical pesticides. The study was for a 2 yr-period; a minimum of three 0.0004-ha sections of row, or 0.0001-ha sections in broadcast or drilled crops were sampled weekly. In addition, sweeps with a net and some other sampling techniques were used in some crops.Litomastix truncatella (Dalman),Meteorus autographae Muesebeck, andVoria ruralis Fallen were the most important larval parasitoids recovered from cabbage loopers;Trichogramma spp. were by far the major egg parasitoid. Parasitization of eggs and larvae in crucifers (Brassica oleracea L.) ranged from 0 to 55% and 0 to 100%, respectively, and was generally highest during the spring and fall. Parasitization of cabbage looper immatures was highest in tomatoes (Lycopersicon esculentum Mill.). Parasitism by the parasitoid complex for the soybean looper larvae was high but eggs in soybeans were seldomly attacked by parasitoids. Tobacco budworm eggs were rarely parasitized in tobacco but were frequently parasitized in tomatoes and okra (Abelmoschus esculentus (L.)Medik). Parasitization of tobacco budworm larvae in tobacco was usually over 50% and was mostly byCardiochiles nigriceps Viereck; fewC. nigriceps were found from the other plants.     

The effect of N-nitroso-N-methylurea,buthylmethane sulphonate and X-rays on the germination and production of chlorophyll mutations in einkorn wheat

Grains ofTriticum monococcum L. var.sofianum Körn. were treated with 0·1mm, 0·2mm and 0·3mm solutions of N-nitroso-N-methylurea (MNH), with 0·03m solution of buthylmethane sulphonate (BMS) and with X-rays in doses of 5 000r and 10 000r. The germination and development of individual colors of chlorophyl mutants were observed by the system developed byLamprecht (1960). All the mutants induced were classified according to their color changes into three main categories-homogenous unicolor, homogenous multicolor and heterogenous multicolor. In the last type the colors of individual leaves of the same plant varied. Anthocyanin mutations “albina” and “albino-transvirgata” sometimes coincided with the chlorophyll mutations. Some chlorophyll mutations showing complicated groups of colors appeared which were beyond the scale of classification by ordinary systems. The largest proportion in the spectrum of chlorophyll mutations, induced by MNH and X-rays was occupied by mutations of thealbina type. The broadest mutation spectrum in our experiments was induced by the application of 0·3mm MNH. The doses of X-rays used induced relatively higher numbers ofalbina-type chlorophyll mutations than MNH and BMS. In our experiments we succeeded in inducing on medium size samples ofTriticum monococcum L. var.sofianum Körn not only almost all types of chlorophyll mutations, induced byFujii (1960, 1962) andMatsumura (1960), but in addition also a great number of other even more complicated chlorophyll mutations, which have never been previously described inTriticum monococcum. L.     

Hydrolysis of the Leu-Gly bond of phenylazobenzyl-oxycarbonyl-l-Pro-l-Leu-Gly-l-Pro-D-Arg (a substrate of microbial collagenases) by treponemes isolated from the subgingival plaque of periodontitis patients

Cell extracts prepared from several oral treponemes isolated from the subgingival plaque of periodontitis patients showed high enzyme activity toward phenylazobenzyl-oxycarbonyl-l-prolyl-l-leucylglycyl-l-prolyl-d-arginine (a compound used as a substrate for microbial collagenases). One major enzyme hydrolyzing this substrate at the Leu-Gly bond only was partially purified from an unspeciated treponeme (strain US),Treponema denticola ATCC 35405, and 29 different clinical isolates ofT. denticola. TheTreponema US enzyme also hydrolyzed furylacryloyl-l-leucylglycyl-l-prolyl-l-alanine (another substrate of bacterial collagenases) at the Leu-Gly bond. This enzyme also hydrolyzed various collagens and collagen-derived peptides. These treponemal proteases were sensitive to metal chelators andp-chloromercury compounds. The results indicate that human oral treponemes contain enzymes that readily hydrolyze in chromogenic protease substrates the Leu-Gly bond only that is the cleavage site of these substrates also by “true” microbial collagenases.     

Chromosome counts of some Cuban Angiosperms

Chromosome numbers are reported for 14 species collected in Cuba. The first chromosome records are reported forAlbizzia cubana Britton etWilson (2n=26),Atkinsia cubensis (Britton etWilson)Howard (2n=26),Caesalpinia violacea (Mill.)Standl. (2n=24),Colubrina ferruginosa Brongn. (2n=24). Chromosome numbers of the following species were confirmed:Albizzia lebbeck (L.)Benth. (2n=26),Canavalia maritima (Aubl.)Thouars (2n=22),Casuarina equisetifolia Forst. (2n=18),Cedrela mexicana M.J. Roem. (2n=56),Delonix regia (Bojer)Raf. (2n=28),Guazuma tomentosa H.B.K. (2n=16),Lysiloma bahamense Benth. (2n=26),L. latisiliqua (L.)Benth. (2n=26),Samanea saman (Jacq.)Merrill (2n=26),Thespesia populnea (L.)Soland (2n=26).     

Biology ofPyemotes parviscolyti [Acarina: Pyemotidae]

Pyemotes parviscolyti Cross & Moser is phoretic only onPityophthorus bisulcatus Eichhoff; it attacks all stages of this insect except the adult. Females, which contain little or no venom, prey on other scolytids if galleries overlap. Males copulate with females ofPyemotes ventricosus Newport and vice versa, but only males of the mother species result. Copulation withPyemotes scolyti Oudemans was not successful.