Influence of the inoculum size ofLactobacillus plantarum on the competition withEnterobacter cloacae

Summary The size of the inoculum ofLactobacillus plantarum or its natural density, appears to be of predominant importance in the exclusion ofEnterobacter cloacae in mixed fermentations, such as ensilage. In a liquid medium, simulating adverse silage conditions, an initial density ofL. plantarum at least twice that ofE. cloacae was found necessary in order to obtain a succesful silage.     

Electrical augmentation of the antimicrobial activity of silver-nylon fabrics

We measured the silver levels producedin vitro by three silver-coated fabrics, and the resulting antimicrobial effect onPseudomonas aeruginosa,Staphylococcus aureus, andCandida albicans. A significant enhancement of the fabrics' antimicrobial effect was achieved by the passage of weak DC currents, which cause increased liberation of silver ions. The antimicrobial effectiveness of each fabric depended on its textile characteristics. Thus, a silver-coated fabric could potentially serve as an antimicrobial dressing by continuously releasing silver ions into a wound either by passive dissociation or through electrical stimulation.Running Title: Antimicrobial Activity of Silver-Nylon     

A self-referential model for the formation of the genetic code

A model for the formation of the genetic code is presented where protein synthesis is directed initially by tRNA dimers. Proteins that are resistant to degradation and efficient RNA-binders protect the RNAs. Replication becomes elongational producing poly-tRNAs from which the mRNAs and ribosomes are derived. Attributions are successively fixed to tRNAs paired through the perfect palindromic anticodons, with the same bases at the extremities (5′ANA: UNU 3′; GNG: CNC; principal dinucleotides, pDiN). The 5′ degeneracy is then developed. The first pairs to be encoded correspond to the hydropathy correlation outliers (Gly-CC: Pro-GG and Ser-GA: Ser-CU) and to the sector of homogeneous pDiN, composed by two pyrimidines or two purines. These amino acids are preferred in the N-ends of proteins, stabilizers of proteins against catabolism and strong RNA-binders. The next pairs complete the sector of homogeneous pDiN (Asp, Glu-UC: Leu-AG and Asn, Lys-UU: Phe-AA). This set of nine amino acids forms the protein cores with the predominant aperiodic conformation. Next enter the pairs with mixed pDiN (one purine and one pyrimidine), the RY attributions composing the protein N-ends and the YR attributions the C-ends. The last pair contains the main punctuation signs (Ile, Met, iMet-AU: Tyr, Stop-UA). The model indicates that genetic information emerged during the process of formation of the coding/decoding system and that genes were defined by the proteins. Stable proteins constructed the nucleoprotein system by binding to the RNAs that produced them. In this circular rationale, genes are memories in a metabolic system for production of proteins that stabilize it. The simplicity and the highly deterministic character of the process suggest that the Last Universal Common Ancestor populations could be composed, in early stages, of lineages bearing similar genetic codes.     

Transformation of the coryneform bacterium cellulomonas flavigena with plasmid DNA via electroporation

Summary Using electroporation we have transformed Cellulomonas flavigena with a shuttle vector (pJA85) derived from the E. coli plasmid pUC8 and the Brevibacterium lactofermentum plasmid pULRS8. Upon transformation this plasmid was found to be stable, not to undergo detectable deletion, and to express antibiotic resistance markers originating in Brevibacterium.     

Improved L-lysine production by the amplification of theCorynebacterium glutamicum dapA gene encoding dihydrodipicolinate synthetase inE. coli

Summary ThedapA gene (L-2,3-dihydrodipicolinate synthetase: DHDP synthetase) ofCorynebacterium glutamicum JS231, a lysine overproducer, cloned and subcloned inE. coli/C. glutamicum shuttle vector pECCG117 was used to transformE. coli threonine producer and threonine and lysine coproducer. The plasmid pDHDP5812 carryingdapA gene ofC. glutamicum led to increase in lysine production in theseE. coli strains. Threonine and lysine co-producerE. coli TF1 with pDHDP5812 produced lysine with small amount of threonine. The DHDP synthetase activity ofE. coli TF1 carrying pDHDP5812 showed high resistance toward inhibition by lysine.     

An assessment of the cultural capabilities of protoplasts of some wild species of linum

Protoplasts of several wildLinum species were isolated enzymatically from roots, hypocotyls and cotyledons of seedlings, and also from theirin vitro grown shoots and cell suspension cultures. When cultured all these protoplasts divided to produce callus but only good plant regeneration capability was evident in the case ofLinum lewissii and to a much lesser extent forL. strictum. Only rhizogenesis was observed withL. alpinum, L. narbonense, L. grandiflorum andL. altaicum. The high plant regeneration capacity ofL. lewissii from protoplast -derived tissues ofin vitro shoots and cell suspension cultures makes this species an attractive experimental system for somatic genetic manipulation.Abbreviations BAP benzylaminopurine - NAA -naphthaleneacetic acid - CPW cell and protoplast wash solution - gFW gram fresh weight On leave from Department of Crop Sciences University of Alexandria Egypt     

Structural and SAXS analysis of the budding yeast SHU-complex proteins

In Saccharomyces cerevisiae, four proteins, Shu1, Shu2, Psy3 and Csm2, form a stable SHU-complex both in vivo and in vitro. These proteins are involved in the early stages of the homologous recombination DNA damage repair process. In this paper, the crystal structure of the Psy3–Csm2 sub-complex is presented at 1.8 Å resolution and successfully fitted into our small angle X-ray scattering (SAXS) data of the SHU-complex. Taken together with our electrophoretic mobility shift assay (EMSA) results, a model is proposed for the SHU–protein complex coupled with DNA.Structured summary of protein interactions:PSY3 and CSM2 bind by X-ray crystallography (View interaction) PSY3, CSM2, Shu 1 and Shu 2 physically interact by x ray scattering (View interaction)     

Regeneration of plants from callus tissue of the pasture legume Centrosema brasilianum

Plants were regenerated from leaf explants of Centrosema brasilianum cultured in vitro. Callus and buds were produced on Murashige and Skoog medium (MS), 0.8% agar, 0.1 mg/l NAA and 1 mg/l BAP. Regeneration of multiple shoots was achieved by transferring callus onto fresh medium containing 0.01 and 1 mg/l of NAA and BAP, respectively. Shoots formed roots upon transfer to MS with 0.01 mg/l NAA. Plantlets were succesfully transferred to soil. Leaf-derived calli of Centrosema arenarium, C. macrocarpum, C. pascuorum, C. pubescens, and C. virginianum did not produce shoots when cultured in vitro.     

Expression of the threonine operon fromEscherichia coli inBrevibacterium flavum andCorynebacterium glutamicum

Summary The threonine operon fromEscherichia coli was cloned in plasmid pBR322, subcloned into the shuttle vector pCEM300 and the resulting recombinant plasmid was transferred intoBrevibacterium flavum andCorynebacterium glutamicum. The expression ofE. coli threonine genes in these coryneform bacteria was demonstrated by complementing thethrA andthrB mutations and by assaying homoserine dehydrogenase activity.     

Conformations of cyclic 3',5'-nucleotides. Effect of the base on the synanti conformer distribution

The furanose and the phosphate rings of cyclic 3′,5′-nucleotides are locked in the ₄T³ and chair conformations respectively. The only variable which shows major conformational flexibility in these molecules is the rotation about the glycosyl bond which describes the orientation of the base relative to the sugar-phosphate bicyclic system. The glycosyl torsion angle has been analyzed for cyclic nucleotides with different purine and pyrimidine bases by use of conformational energy calculations. The results indicate that all the pyrimidine bases, U, T and C show a very strong energetic preference for the anti range of conformations. The calculations predict that among cyclic 3′,5′-purine nucleotides cyclic GMP and cyclic IMP favor the syn conformation to the anti by 95:5 and 70:30 respectively, while cyclic AMP shows a preference for the anti conformation to syn by 70:30. Thus the purines show a greater probability for the syn conformation than the pyrimidines in cyclic 3′,5′-nucleotides.     

An investigation into the 19-hydroxylation of androstenedione,cortexolone and progesterone by selected fungi

Summary An investigation was performed to recognise a fungus capable of 19-hydroxylating the steroids androstenedione, cortexolone and progesterone, in high yield with little or no side reactions. The fungi selected for study belong to a group which has been previously reported to possess 19-hydroxylation ability. Pellicularia filamentasa f. sp. microsclerotia IFO 6298 and P. filamentosa f.sp. sasakii IFO 5254 both 19-hydroxylated cortexolone, but 19-hydroxylated products were not detected using either androstenedione or progesterone as substrate. When 19-hydroxylating cortexolone, both strains of P. filamentosa produced 11-hydroxycortexolone as a by-product, but P. filamentosa f.sp. microsclerotia IFO 6298 was superior in terms of 19-hydroxylation and the relative amounts of the two products.     

Cloning a mutatedtrp operon for the biosynthesis of an antibiotic agent

Summary Plasmid pCG8 containing a mutatedtrp operon was constructed and introduced into anEscherichia coli host for increasing tryptophan biosynthesis. Since tryptophan is a biosynthetic precursor of pyrrolnitrin, the mutated trp operon was inserted into aPseusdomonas vector pME290 to obtain the plasmid pCG12, and then was introduced intoPseudomonas pyrrocinia for enhancing the pyrrolnitrin synthesis. Data showed that the production of pyrrolnitrin of the transformed strain was five-times greater than that of the parental strain.     

Brine organisms and the question of habitat-specific adaptation

Among the well-known ultrasaline terrestrial habitats, the Dead Sea in the Jordan Rift Valley and Don Juan Pond in the Upper Wright Valley represent two of the most extreme. The former is a saturated sodium chloride-magnesium sulfate brine in a hot desert, the latter a saturated calcium chloride brine in an Antarctic desert. Both Dead Sea and Don Juan water bodies themselves are limited in microflora, but the saline Don Juan algal mat and muds contain abundant nutrients and a rich and varied microbiota, includingOscillatoria,Gleocapsa,Chlorella, diatoms,Penicillium and bacteria.In such environments, the existence of an array of specific adaptations is a common, and highly reasonable, presumption, at least with respect to habitat-obligate forms. Nevertheless, many years of ongoing study in our laboratory have demonstrated that lichens (e.g.Cladonia), algae (e.g.Nostoc) and fungi (e.g.Penicillium,Aspergillus) from the humid tropics can sustain metabolism down to –40°C and growth down to –10°C in simulated Dead Sea or Don Juan (or similar) media without benefit of selection or gradual acclimation. Non-selection is suggested in fungi by higher growth rates from vegetative inocula than spores. The importance of nutrient parameters was also evident in responses to potassium and reduced nitrogen compounds.In view of the saline performance of tropicalNostoc, and its presence in the Antarctic dry valley soils, its complete absence in our Don Juan mat samples was and remains a puzzle.We suggest that adaptive capability is already resident in many terrestrial life forms not currently in extreme habitats, a possible reflection of evolutionary selection for wide spectrum environmental adaptability.     

Lactic acid bacteria active during the fermentation of wheat silage in small scale silos

Summary Wheat was ensiled and periodically analyzed for lactic acid bacteria present. Initially Lactobacillus plantarum, Leuconostoc mesenteroides, Lactobacillus cellobiosus and Streptococcus lactis predominated. After two to four days enterococci including S. faecium and S. bovis were present in high populations as well as Lactobacillus plantarum. It was concluded that mixed populations of enterococci and L. plantarum are active in the successful fermentation of wheat silage.     

Conversion of sugar-beet particles to ethanol by the bacteriumZymomonas mobilis in solid-state fermentation

Summary The possibility of usingZymomonas mobilis as the microorganism, in solid-state fermentation of sugar-beet particles was investigated. The major factors affecting the process were investigated and related to ethanol yield and productivity. Ethanol yield of 0.48 g/g sugar, volumetric productivity of 12 g/L h, and final ethanol concentration of 130 g/L show the good performance ofZ.mobilis in a solid-state fermentation.     

Applicability of the PCR technique in the food testing laboratory: Identification of Listeria monocytogenes

A polymerase chain reaction (PCR) targeting Listeria monocytogenes-specific hlyA gene sequences was evaluated as a tool for the identification of presumptive positive colonies isolated on Palcam or Oxford agar during routine analysis of food samples by culture technique. The PCR correctly identified L. monocytogenes isolates from a large number of samples, and provided a significant saving in the time, labour and cost compared to the traditional biochemical tests.     

Effect of the genes B and Pl on anthocyanin synthesis in maize endosperm culture

B and Pl are two genes involved in anthocyanin biosynthesis in maize (Zea mays) plant tissues. In this work the effect of B and Pl on pigment accumulation is analyzed in endosperm tissues, either cultured in vitro or scraped off from the seed. The results obtained indicate that the two genes play a different role in callus pigmentation: B exerts a qualitative change in pigment composition, while Pl controls the rate of pigment accumulation in the callus. Anthocyanin synthesis in all strains analyzed appears to be light independent. Two cases of instability in pigment production arisen in the endosperm cultures are described and discussed in relation to epigenetic variation in secondary metabolite content in plant tissue culture.Abbreviations BEAF Benzene/ethyl acetate/formic acid (40:10:5) - 2-4D 2,4-dichlorophenoxyacetic acid - Wi initial weight - Wt total weight     

The effect of the H2 partial pressure on the metabolite pattern ofLactobacillus casei,Escherichia coli andClostridium butyricum

Summary The metabolite pattern of batch cultures ofLactobacillus casei LMG 6400,Clostridium butyricum LMG 1213t1 andEscherichia coli LMG 2093 was effected only for the latter organism when the H₂ partial pressure was below 1 atmosphere: high hydrogen partial pressures increased the formate formation, low pressures gave rise to increased acetate production and higher cell yields.     

Factors affecting the use of chloramphenicol acetyltransferase as a marker for Brassica genetic transformation

The CAT gene which codes for the enzyme chloramphenicol acetyltransferase was found to be ineffective as a reporter gene in cells and tissues of Brassica species. High levels of endogenous CAT activity were found to be widespread among this genus and did not appear to be distributed in a tissue- or cell-specific manner. Moreover, the presence of an inhibitor of CAT activity was discovered in Brassica napus and Brassica juncea. This inhibitor appeared to act selectively on bacterial CAT in transgenic plants. These findings provided an explanation for difficulties experienced in the detection of transgenic CAT activity in B. napus.     

In situ detection of transposition of the maize controlling element (Ac) in transgenic soybean tissues

The development of a transposon mutagenesis system in soybean would aid in the isolation of unknown genes. The maize controlling element (Ac) has, therefore, been introduced into the soybean (Glycine max (L.) Merr.) genome byAgrobacterium-mediated transformation.Ac was inserted into the untranslated leader region of the bacterial ß-glucuronidase gene (GUS) such that the excision ofAc resulted in restoration of the GUS gene activity. Excision events of theAc element were monitored by detecting blue cells or sectors in transgenic soybean tissues. Using the GUS gene assay and with hybridization data, we have demonstrated that theAc element transposes in transgenic soybean calli, leaves, stems, and roots.