Abstract: A survey was conducted on the isolation and characterization of bacteriocin-producing lactic acid bacteria in soil. Forty-two acid-producing bacterial strains were isolated from 55 soil samples collected in Yamanashi prefecture, Japan. Investigation of antibacterial activities of isolates revealed that three isolates, Lactobacillus animalis C060203, Enterococcus durans C102901 and Leuconostoc mesenteroides subsp. mesenteroides C060204, showed antibacterial activities against the indicator strain of Lactobacillus sakei JCM 1157T. Bacteriocin from Enterococcus durans C102901 showed different characteristics from the known durancin L28-1A, produced by Enterococcus durans L28-1. Furthermore, this is the first report of a bacteriocin being produced by Lactobacillus animalis. Viewing from the species, bacteriocins from strains C102901 and C060203 showed high possibilities for the novel substances. These significant findings suggest that soil may be a common source for the isolation of novel bacteriocin-producing lactic acid bacteria.

Abstract: Chlorophenols (CPs) are major environmental pollu-tants discharged from pulp and paper mills, tanneries,distilleries, dye and paint manufacturing and pharma-ceuticals industry (Crosby, 1981). The major source ofCPs is as by-products when chlorine is used forbleaching of pulp and for disinfections of drinkingwater. Therefore, large amounts of chlorinated aro-matic compounds, including CPs, have been dis-charged into the environment (Steinle et al., 1998).Among chlorophenols, pentachlorophenol (PCP) is awide spectrum biocide with applications in agriculture,industry and public health (Bevenue and Beckman,1967). PCP a highly toxic pesticide has been widelyused in disinfectants and preservatives (Rao, 1978).The U.S. Environmental Protection Agency regulatesPCP as a priority pollutant and also considers 1.0mg/Lof PCP hazardous for land disposal (Freeman, 1989;Sittig, 1981).PCP is expected to be recalcitrant to aerobicbiodegradation because it is highly chlorinated and, ingeneral, aromatic compounds with higher amounts ofchlorine are more resistant to biodegradation (Anan-darajah et al., 2000). In addition, this compound is veryharmful to microorganisms because it destroys mem-brane function due to its ability to uncouple oxidativephosphorylation (Copley, 2000; Ito and Ohnishi, 1982).In spite of these adverse properties, several microor-ganisms have been isolated from sites contaminatedwith PCP and their metabolite biodegradation path-ways have been elucidated. In addition to this, differ-ent researchers have reported the PCP degrading microorganism (bacteria) from the natural environ-ment. A number of microorganisms,

Abstract: Our study aims to investigate and describe the epidemiology of the intestinal carriage of ESBL-PS in intensive care units of five Lebanese hospitals and to analyze the potential risk factors for the acquisition of these strains At the same time, we intend to determine the patterns of susceptibility of these strains, exploring therefore the availability of alternative treatment One thousand, four hundred forty-two fecal samples were collected between January 1, 2003 and March 31, 2003 from 378 patients admitted to the ICUs of five Lebanese tertiary care general hospitals located in different areas of Lebanon ESBL production was detected by the double disk synergy test and antibiotic susceptibility of ESBL-producing strains as well as minimum inhibitory concentrations were determined A paired case-control study was undertaken to identify risk factors for carriage of ESBL-PS One hundred eighteen strains isolated from 72 subjects were identified as ESBL producers, including 95 (805%) E coli, 16 (136%) Klebsiella pneumoniae, and 7 (56%) Enterobacter cloacae A higher rate of multiple ESBL-PS carriage was described among these acquisition cases (21 double carriages and 3 triple carriages of ESBL-PS compared to only 1 double carriage of ESBL-PS at admission) In general, similar trends of susceptibility were observed in the different hospitals As expected, the lowest MIC was observed with imipenem for all E coli, Klebsiella, and Enterobacter isolates Ciprofloxacin, followed by trimethoprim-sulfamethoxazole seem to be associated with the lowest susceptibility In vitro susceptibility to cefoxitin for all isolates was 746%; more resistance was associated to ceftazidime (907%) than to cefotaxime (697%) Our data agree with other national and international reports showing the increase in ESBL-PS carriage in ICU patients They demonstrate the endemic character of this carriage in Lebanese hospitals and the important risk factors including immunosuppression and evidence of ESBL infection The highly resistant profile of ESBL-PS to antimicrobial agents available for treatment reflects the severity of this issue The significance of this study resides in the direct correlation between our results and the nationwide increase in multi-drug resistant bacteria and the continuous change in bacterial resistance epidemiology Our data may have an important impact on infection control policies in hospitals and on treatment of infectious diseases

Abstract: A thermophilic, spore-forming bacterial strain L1T was isolated from hot compost “Pomigliano Environment” s.p.a., Pomigliano, Naples, Italy. The strain was identified by using a polyphasic taxonomic approach. L1T resulted in an aerobic, gram-positive, rod-shaped, thermophilic with an optimum growth temperature of 68°C chemorganotrophic bacterium which grew on hydrocarbons as unique carbon and energy sources and was resistant to heavy metals. The G+C DNA content was 43.5 mol%. Phylogenetic analysis of 16S rRNA gene sequence and Random Amplified Polymorphic DNA-PCR (RAPD-PCR) analysis of L1T and related strains showed that it forms within Geobacillus toebii, a separate cluster in the Geobacillus genus. The composition of cellular fatty acids analyses by Gas-Mass Spectroscopy differed from that typical for the genus Geobacillus in that it is lacking in iso-C15 fatty acid, while iso-C16 and iso-C17 were predominant. Isolates grew on a rich complex medium at temperatures between 55–75°C and presented a doubling time (td) of 2 h and 6 h using complex media and hydrocarbon media, respectively. Among hydrocarbons tested, n-decane (2%) was the more effective to support the growth (1 g/L of wet cells). The microorganism showed resistance to heavy metal tested during the growth. Furthermore, intracellular α-galactosidase and α-glucosidase enzymatic activities were detectable in the L1T strain. Based on phenotypic, phylogenetic, fatty acid analysis and results from DNA-DNA hybridization, we propose assigning a novel subspecies of Geobacillus toebii, to be named Geobacillus toebii subsp. decanicus subsp. nov., with the type strain L1T (=DSM 17041=ATCC BAA 1004).

Abstract: Forty-nine strains belonging to the genus Gluconobacter were re-examined with respect to their species identification based on the sequences of the 16S rDNA and 16S-23S rDNA internal transcribed spacer regions (ITS). A phylogenetic tree constructed from the 16S rDNA sequences indicated the presence of five clusters corresponding, respectively, to the major five species of the genus Gluconobacter, namely G. albidus, G. cerinus, G. frateurii, G. oxydans (type species), and G. thailandicus. The type strain of G. asaii, NBRC 3276T (T=type strain) was included in the G. cerinus cluster, which is consistent with the report that G. asaii is a junior subjective synonym of G. cerinus. Existence of the G. albidus, G. cerinus, G. frateurii, G. oxydans, and G. thailandicus clusters was also recognized by the ITS sequence analysis. Both sequence analyses revealed that the G. cerinus and G. frateurii clusters were heterogeneous. The G. cerinus cluster comprised three strains of G. cerinus and one strain of G. frateurii, while the G. frateurii cluster included ten strains of G. frateurii, three of G. cerinus, and eleven of G. oxydans. These results suggest that phenotypic differences among Gluconobacter species are ambiguous and the species definition must be re-evaluated. The 16S rDNA and ITS sequences determined in this study are valuable for the identification and phylogenetic analysis of Gluconobacter species.

Abstract: no cross-reactivity) proteins known as botulinum neurotoxin (serotypes A–G) and tetanus neurotoxin (NT) have apparent similar structures and similar structurefunction relationships (Humeau et al., 2000; Schiavo et al., 2000). These proteins do not appear to serve any physiological function for the producer anaerobic bacteria Clostridium botulinum, C. baratii, C. butyricum, C. argentinense and C. tetani, nor to be essential for their survival (Allen et al., 1999; Minton, 1995). Are the NTs vestiges or relics of evolution? Or is the ancestral NT gene (Collins and East, 1998; Henderson et al., 1997; Popoff and Marvaud, 1999) still evolving to acquire a “useful existence”? The NT genes, seemingly superfluous for the survival and reproduction of C. botulinum, express the most toxic poison in nature that does not kill any cell. This conundrum is considered with some conjectures, viewed from the published structural and functional properties of the 150-kDa proteins. Assessment of the literature provided rationale to propose a consideration of the NT as a polyprotein harboring viral protease (the 50-kDa light chain) and to provide a perspective on the evolution of the NT into the present state where 8 NTs (botulinum plus tetanus) cleave 3 neuronal and 1 non-neuronal proteins at 7 sites. Clostridial NTs are polyproteins harboring viral metalloproteinases—a proposal: Each NT synthesized as a 150-kDa single chain protein shapes into three clearly visible (X-ray diffraction of crystals) structural domains each of 50 kDa (Lacy et al., 1998; Swaminathan and Eswaramoorthy, 2000). Proteolytic cleavage(s) at the two junctions between the three structural-functional domains is a facile process as is their ensuing separation(s) (Prabakaran et al., 2001, and refs. therein). The selective and relative susceptibilities of these junctional segments indicate that these segments are mere hinges, as is evident from the 3-dimensional structures of type A and B botulinum NTs, acting as purely provisional links between the three structurally independent domains. Each of the three 50-kDa domains of the 150-kDa NTs performs its individual function without the presence of the other two. The 50-kDa C-terminal domain (the binding domain) mediates NT’s binding to the receptors on the presynaptic membrane of neuromuscular junctions. The adjacent 50-kDa domain forms channels in the endosomal membrane (channel former or transmembrane domain) and promotes insertion/passage of the Botulinum neurotoxins: Perspective on their existence and as polyproteins harboring viral proteases

Abstract: A fungus producing magenta was isolated from cellulosic material by visual observation on Czapek's agar media and the product was conventionally analyzed. The fungal strain that produced magenta pigment was closely related to Phoma herbarum. The type of fibers added to Czapek's medium influenced which pigments were produced. Mycelia attached to the surface of nylon-6 and excreted magenta pigment into the fibers. The pigment structure was partially determined. This is the first report of the production of magenta pigment by a microorganism specifically in the presence of nylon-6 fibers, via an unknown mechanism. This phenomenon raises the question of why and how the fungus disperses the pigment inside the fiber and suggests that fabrics can be dyed using microorganisms.

Abstract: In the Escherichia coli pgsA null mutant, which lacks the major acidic phospholipids, the Rcs phosphorelay signal transduction system is activated, causing thermosensitive growth. The mutant grows poorly at 37°C and lyses at 42°C. We showed that the poor growth at 37°C was corrected by disruption of the rcsA gene, which codes for a coregulator protein that interacts with the RcsB response regulator of the phosphorelay system. However, mutant cells still lysed when incubated at 42°C even in the absence of RcsA. We conclude that the activated Rcs phosphorelay in the pgsA null mutant has both RcsA-dependent and -independent growth inhibitory effects. Since the Rcs system has been shown to positively regulate the essential cell division genes ftsA and ftsZ independently of RcsA, we measured cellular levels of the FtsZ protein, but found that the growth defect of the mutant at 42°C did not involve a change in the level of this protein.

Abstract: This study describes the amplification, localization, and sequence analysis of a hemolysin gene from type strain V. campbellii NBRC 15631--the first report of a full-length hemolysin gene for the species. An amplicon ( approximately 600 bp) of polymerase chain reaction performed using V. campbellii DNA template and primers previously designed to target a fragment of V. harveyi hemolysin gene (vhh) was shotgun-cloned and sequenced, generating 576 bp nucleotide sequences of the V. campbellii hemolysin gene. PCR primers designed based on these initial sequences were used to amplify a 551-bp V. campbellii hemolysin gene fragment that was used as probe in Southern hybridization, which localized the complete hemolysin gene within a 3.5-kb HindIII restriction fragment of the V. campbellii genomic DNA. To obtain the remaining DNA sequences upstream and downstream of the 576-bp hemolysin gene sequences, inverse PCR was performed using a self-ligated (circularized) V. campbellii HindIII restriction fragment as the template and PCR primers designed to amplify flanking regions of the 576-bp gene fragment. Nucleotide sequences from the terminal regions of the 3.1-kb product of inverse PCR provided the flanking sequences, resulting in the complete sequence for the V. campbellii hemolysin gene. A VCH PCR primer set was designed to amplify a 1.3-kb region containing the entire hemolysin gene even from other V. campbellii strains, which was sequenced to confirm the V. campbellii hemolysin gene sequence. An open reading frame (ORF) of 1,254 bp (designated as vch) was identified, sharing 79% sequence identity with V. harveyi hemolysin gene vhh, representing 262 base substitutions between V. campbellii and V. harveyi. The deduced amino acid sequence of V. campbellii hemolysin (VCH) shows homologies to the V. harveyi hemolysin (VHH), thermolabile hemolysin of V. parahaemolyticus, proteins such as phospholipase of V. vulnificus and lecithinases of V. mimicus and V. cholerae. The VCH primer set did not produce any amplicon in PCR using V. harveyi DNA, and may therefore be used to distinguish environmental strains of V. campbellii from V. harveyi.

Abstract: Three hundred and eight presumed enterococcal isolates were recovered from Bryndza, a soft sheep milk cheese. The cheese samples were obtained from five different commercial distributors in Slovakia and were taken at three different seasonal intervals. All isolates were identified to the species level using genotypic tools. Species-specific PCR using ddl genes highlighted the predominance of Enterococcus faecium (176 isolates) and assigned 50 isolates to the species Enterococcus faecalis. The remaining 82 isolates were classified using repetitive element sequence-based polymerase chain reaction (PCR) with primer (GTG)5‐(GTG)5-PCR, in combination with phenylalanyl-tRNA synthase gene (pheS) sequence analysis and by whole-cell protein analysis (SDS-PAGE). These strains were identified as Enterococcus durans (59 strains), Enterococcus italicus (8 strains), Enterococcus casseliflavus (3 strains), Enterococcus gallinarum (3 strains), Enterococcus hirae (1 strain), and 8 strains were members of the species Lactococcus lactis. Of the seven enterococcal species isolated, three of them, E. durans, E. faecalis and E. faecium were present in all samples studied, with E. faecium as the predominant one. The precise identification of enterococci in Bryndza cheese is an essential step in the process of evaluation of their functional properties which will be further studied and assessed.

Abstract: We have investigated the molecular phylogeny of cold-seep sediments obtained from the Nankai Trough, at depths of about 600, 2,000, and 3,300 m, and compared the microbial diversity profiles of those sediments samples. The γ-Proteobacteria that might function as sulfide oxidizers and the symbiotically related δ-Proteobacteria which might function as sulfate reducers were identified amongst the bacteria from all depths of the sediments. However, anoxic methane oxidizing archaea (ANME) and methanogens were only found in the 600 m deep sediments. These results indicated that the cold-seep microbial sulfur circulation system could be functioning in the shallow seep sediment at a depth of 600 m and the microbial activities at these sites might be more dynamic than at other deeper cold-seep sites.

Abstract: Seven strains of ballistoconidiogenous yeasts that contain xylose, form Q-10 ubiquinone, propagate by budding and don't produce stalk conidia were isolated from plant leaves collected in Thailand and were found to represent two new species. The taxonomic properties of the two species coincided with the genus Bullera so they are described as Bullera koratensis sp. nov. and Bullera lagerstroemiae sp. nov. In phylogenetic trees based on the nucleotide sequences of 18S ribosomal DNA and the D1/D2 domain of 26S rDNA, these two species are located in the Trichosporonales clade (Cryptococcus humicola-Trichosporon lineage).

Abstract: The antimycobacterial activity (both in vitro and in vivo) and DNA gyrase inhibition of newly synthesized fluoroquinolone derivatives were tested against Mycobacterium tuberculosis $H_{37}$Rv and Mycobacterium smegmatis, respectively. Among the synthesized compounds, compound F11 was found to exhibit the most potent in vitro antimycobacterial activity with a MIC value of 0.78 μg/ml, and a selectivity index of more than 80 while not being cytotoxic to the Vero cell line up to 62.5 μg/ml. When evaluated for in vivo antimycobacterial activity, compound F11 demonstrated a paramount decrease of bacterial load in lung and spleen tissues compared to the control and better than the standard drug ciprofloxacin.

Abstract: This study was focused on obtaining the complete gene sequence of the toxR gene in V. harveyi by using toxR-targeted PCR to amplify 5′ and 3′ regions flanking the 576-bp Vibrio harveyi (NBRC 15634) toxR gene fragment previously amplified using degenerate PCR. To obtain the 5′ flanking sequences, a forward PCR primer (VhtoxRpv) was designed based on known sequences upstream of toxR in V. parahaemolyticus and V. vulnificus. The reverse primer (VctoxR2R) was based on the sequence of the 576-bp Vibrio harveyi toxR fragment. The resulting 750-bp amplicon was sequenced, providing the 5′ sequences of the V. harveyi (NBRC 15634) toxR gene. The 3′ flanking region was amplified using a primer pair toxRS1 and toxRS2 based on V. parahaemolyticus and V. vulnificus toxR and toxS, resulting in a 900-bp amplicon that contained the remaining 3′ sequences of the V. harveyi NBRC 15634 toxR. This paper reports, for the first time, a complete 882-bp nucleotide sequence for toxR in Vibrio harveyi. Sequence analysis and alignment revealed that the complete toxR gene in V. harveyi shares 87% sequence similarity with toxR of V. parahaemolyticus, 84% similarity with V. fluvialis, 83% with V. vulnificus and partial sequence of V. campbellii. The phylogenetic trees revealed wider divergence in toxR compared to 16S rRNA genes, so that V. harveyi could easily be distinguished from V. campbellii and V. parahaemolyticus.

Abstract: Bacteriocins ST194BZ and ST23LD, produced by Lactobacillus plantarum, inhibit Gram-positive and Gram-negative bacteria. Images obtained by atomic force microscopy showed clear signs of membrane damage of Lactobacillus sakei, accompanied by the leakage of DNA and β-galactosidase. Adsorption of the bacteriocins to cells was increased when cells were treated with buffers at pH values above neutral. An increase in bacteriocin ST194BZ adsorption to cells of Enterococcus sp. and L. sakei was observed with an increase in incubation temperatures, but at different rates for the two species. Treatment of the two species with various inorganic salts and solvents gave different results regarding the adsorption of the two bacteriocins. In general, pre-treatment of the two sensitive cells with Triton X-100, Triton X-114 and chloroform increased the adsorption of the two bacteriocins. Increased adsorption of bacteriocin ST23LD to L. sakei was recorded when the cells were pre-treated with Tris and NH4-citrate. Treatment of Enterococcus sp. and L. sakei with Na-EDTA and SDS decreased the adsorption of the two bacteriocins. Variable results were recorded with inorganic salts.

Abstract: The research was focused on the multiplex polymerase chain reaction (PCR) differential detection of shrimp pathogens Vibrio harveyi, Vibrio campbellii and isolates from a variant strain of Vibrio (referred to as Philippine Vibrio isolates in this study) exhibiting characteristics distinct from these two species. Sequence alignment of the hemolysin gene from type strains Vibrio harveyi (NBRC 15634) and Vibrio campbellii (NBRC 15631), as well as 10 variant Philippine Vibrio isolates, was performed in order to design a set of hemolysin-targeted primers for the specific detection of the Philippine Vibrio isolates. Primer PNhemo amplified a 320-bp hemolysin gene fragment of the Philippine Vibrio isolates in PCR using 65 degrees C annealing temperature, but did not amplify the target gene fragment in type strains V. harveyi and V. campbellii. Another new primer (VcatoxR) targeting the toxR gene was designed for the specific detection of type strain V. campbellii under stringent 65 degrees C annealing temperature. PCR using VcatoxR primer resulted in the specific amplification of a 245-bp V. campbellii toxR fragment. The simultaneous use of three primer sets in PCR, including PNhemo and VcatoxR (the two new primers designed in this study), and a primer VhtoxR (previously reported for the specific detection of V. harveyi), resulted in differential profiles with 390-bp, 245-bp, and 320-bp amplicons for V. harveyi, V. campbellii, and variant Philippine Vibrio isolates, respectively. Presence of all three types of Vibrio shrimp pathogens in the sample could be detected with a multiplex PCR profile containing all the expected size amplicons.

Abstract: The SigB concentrations in clinical isolates of Staphylococcus aureus were measured to examine their correlation with the antibiotic resistance. The SigB concentrations in methicillin-resistant S. aureus (MRSA) were higher than in the control strain, N315, and many of methicillin-susceptible S. aureus (MSSA). Sequencing analyses of the sigB genes revealed that the strains exhibiting the high SigB concentrations have three amino acid substitutions in SigB: I11V, D141N, and Q256K. Further analysis using isogenic mutants demonstrated that D141N (or both D141N and Q256K) is essential to maintain the high SigB concentration. These substitutions affected the UV tolerance, but had no effect on the antibiotic resistance. The SigB activity was affected by these substitutions toward the stationary phase, but not during the transient heat shock response.

Abstract: In this study, 129 Turkish high-sugar products were examined in terms of their yeast flora and 73 representative strains were isolated. Yeast isolates were identified at species level by using Apilab Plus (bioMerieux, France), a specific computer program developed for ID 32C strips (bioMerieux, France). While one of the isolates could be identified at genus level as Aureobasidium, 66 of them were identified as 21 species belonging to 8 different genera. The distribution of these isolates were as follows: Candida (38), Rhodotorula (8), Zygosaccharomyces (7), Cryptococcus (6), Saccharomyces (3), Debaryomyces (2), Pichia (1) and Torulaspora (1). Approximately 70% of the isolates were found to have the ability to grow on media with 50% (w/w) glucose. Hence, they were characterized as xerotolerant strains. Although Zygosaccharomyces rouxii is known as the most xerotolerant yeast species, only two strains of Z. rouxii could be isolated from Turkish high-sugar foods. During identification studies, it was observed that ID 32C test strips should certainly be supported by morphological and physiological tests for obtaining more reliable identification results. If not, closely related yeast species such as anamorph and telemorph forms can not be distinguished.

Abstract: Recent advanced studies of genomics and proteomics have revealed the variation and diversity of ribosomal proteins (r-proteins) in different organisms and organelles. Radical free and highly reducing (RFHR) two-dimensional (2-D) electrophoresis is known to be powerful for separating ribosomal proteins that are usually small and basic, and not separated well by standard 2-D electrophoresis. Using the RFHR method, we investigated the protein profile of the Bacillus subtilis ribosomes by a proteomic approach. We found that two L31 paralogue proteins (RpmE and YtiA) showed different temporal expression patterns in the ribosomes. The RpmE protein, which is an L31 variant with a Zn-binding motif, binds one zinc ion at the motif, which is required for stabilization of the protein in the cell. On the other hand, the expression of the ytiA gene, which encodes another L31 variant (YtiA) without the Zn-binding motif, is negatively controlled by the zinc-specific transcriptional repressor Zur and is likely induced by zinc starvation. This article reviews the recent findings that replacement of two types of L31 proteins in the ribosome is controlled by the intracellular zinc concentration.

Abstract: Phylogenetic relationships within the Erythrobasidium clade as a lineage of the urediniomycetous yeasts were examined using partial regions of 18S rDNA, 5.8S rDNA, 26S rDNA, internal transcribed spacers (ITSs), and elongation factor (EF)-1α. Combined data analysis of all segments successfully yielded a reliable phylogeny and confirmed the cohesion of species characterized by Q-10(H2) as a major ubiquinone. Differences in secondary structure predicted for a variable region in 26S rDNA corresponded to major divergences in the phylogenetic tree based on the primary sequence. The common presence of a shortened helix in this region was considered to be evidence of monophyly for species with Q-10(H2), Sakaguchia dacryoides, Rhodotorula lactosa, and Rhodotorula lamellibrachiae, although it was not as strongly supported by the combined data tree. The information on intron positions in the EF-1α gene had potential usefulness in the phylogenetic inference between closely related species.

Abstract: There are two structural profiles in the yeast Golgi. The Golgi of Saccharomyces cerevisiae is composed of a number of vesicular compartments dispersed in the cytoplasm as recognized by a large number of Golgi marker proteins. In contrast, the Golgi of Pichia pastoris was reported to be organized in a small number of stacked cisternae located near the transitional endoplasmic reticulum (tER) sites by electron microscopy and immunofluorescent staining of a few marker proteins. The guanosine diphosphate (GDP)-mannose transporter (GMT) is an essential component in the yeast Golgi apparatus. We isolated an ortholog of the GMT gene of P. pastoris and visualized the gene product by epitope tagging to verify the structural characteristics of the Golgi. The tagged product in P. pastoris cell was observed in rod-like compartments in which Och1 mannosyltransferase was also found and the tER marker Sec12 and Sec13 proteins localized very close to them. The present results add further evidence of the restricted localization of the Golgi in P. pastoris cell.

Abstract: To examine the occurrence in other deep-sea bacteria of two amino acid substitutions (Ala-180 and His-229) in malate dehydrogenase (MDH) found previously in the deep-sea piezophilic Moritella sp. strain 2D2, we cloned and sequenced MDH genes of deep-sea piezophilic Moritella and Shewanella strains isolated from intestinal contents of deep-sea fishes, as well as other Moritella species from deep-sea water and sediments: M. marina, M. japonica, and M. yayanosii. The piezophilic Moritella strains had a Val residue or an Ala residue at position 180 and all the Moritella strains except for one had a His residue at position 229. However, four piezophilic-strain-specific substitutions at positions 103, 111, 229, and 283 were found to be completely conserved in the MDH of the intestinal Moritella strains of deep-sea fishes, indicating the substitutions may be habitat-specific. The piezophilic Shewanella strains had a Val residue and a Gln residue at positions 180 and 229, respectively. However, the MDHs of the Shewanella strains had five piezophilic-strain-specific substitutions at positions 61, 65, 107, 161, and 202. Therefore, the enzymatic strategies for responding to deep-sea high pressure environments of the MDHs between the genera Moritella and Shewanella are potentially different. Moreover, homology modeling shows these substitutions found in the MDHs of both genera except for position 229 in the subunit interface are located on the exposed region of the MDH molecules, indicating the substitutions may be related to the hydration state of the molecules.

Abstract: nine lower eukaryotic phyla, the phylum Apicomplexa, Chlorarachniophyta (Cercozoa), Chlorophyta, Ciliophora, Dinophyta, Euglenophyta (Euglenozoa), Glaucophyta, Heterokontophyta (Heterokonta) and Rhodophyta, have been studied in order to consider the phylogenetic significance of cellular polyamine distributions in early evolution of eukaryotes (Hamana and Matsuzaki, 1982, 1985; Hamana et al., 1990, 2004a, b). In the nine phyla, the three phototrophic phyla Glaucophyta, Rhodophyta and Chlorophyta have plastids by the primary endosymbiosis of a phototrophic prokaryote, cyanobacterium, and multicellular species evolved within Rhodophyta and Chlorophyta (Bhattacharya et al., 2004; Cavalier-Smith, 1998; Falkowski et al., 2004; NCBI website, 2006; Rodriguez-Ezpeleta et al., 2005). The other six unicellular phyla include heterotrophs evolved without the endosymbiosis of cyanobacteria, phototrophs by secondary or tertiary endosymbiotic plastids and non-photosynthetic heterotrophs evolved after the loss of primary endosymbiotic plastids (Bhattacharya et al., 2004; Cavalier-Smith, 1998; Falkowski et al., 2004; NCBI website, 2006; Rodriguez-Ezpeleta et al., 2005). The distribution pattern of three triamines, norspermidine, homospermidine and spermidine, and two tetraamines, norspermine and spermine, was almost phylum-specific among the nine phyla; furthermore, a part of their polyamine components seems to be correlated to their evolutional endosymbiotic process. The remaining three taxa (phyla) among total twelve phyla of lower eukaryotes, include the non-photosynthetic phylum Percolozoa without the endosymbiosys of cyanobacteria and the two photosynthetic phyla Cryptophyta and Haptophyta taken secondary endosymbiotic plastids (Bhattacharya et al., 2004; Cavalier-Smith, 2002, 2003; Falkowski et al., 2004). Cellular polyamines in the three phyla were first analyzed in the present study. Additional polyamine catalogues of some primitive phototrophic members belonging to Glaucophyta and Rhodophyta were determined. Two strains of Glaucophyta, seventeen strains of Rhodophyta, seven strains of Cryptophyta and eight strains of Haptophyta were supplied from IAM, NIES and MBIC, and were cultivated phototrophically in the light using the media designed by the culture collections (IAM Catalogue Strains, 2004; Kasai et al., 2004; MBIC Strain Catalog Algae, 2006). Cyanidioschyzon and Cyamidium species were grown at 37°C. Other red algae and two glaucophytes were grown at 15– 25°C. Seven cryptophytes and eight haptophytes were grown at 10–15°C and 15–20°C, respectively. Five non-photosynthetic species of Percolozoa were Cellular polyamines of lower eukaryotes belonging to the phyla Glaucophyta, Rhodophyta, Cryptophyta, Haptophyta and Percolozoa

Abstract: This study compares the PHB synthase activity of Nostoc muscorum, a N2-fixing cyanobacterium under control (grown in usual BG-11 medium), nitrogen (N) and phosphorus (P) deprivation and chemoheterotrophic conditions. Specific activity of PHB synthase did not depict significant variations in the latter three types of cultures, except for the control one, where a significantly lower activity was recorded. PHB synthase activity was detected only in the soluble fractions of both the control as well as cells incubated under chemoheterotrophic conditions. A Km of 80.2μM DL-β-hydroxybutyryl-CoA and Vmax of 197.5 nmol thiobenzoate (TNB) mg protein−1min−1 were observed for the enzyme. PHB synthase remained insensitive to acetyl-CoA, ATP, NADP, NADPH supplementation under in vitro condition. Addition of acetyl phosphate was found to activate the enzyme and the level of activation was dependent on the concentration of acetyl phosphate supplementation. Inhibition of PHB synthase in 2,3-butanedione supplemented cultures and reactivation following acetyl phosphate addition proved the post-translational control of acetyl phosphate over PHB synthase.

Abstract: Culture supernatant of Bacillus thuringiensis 9816C had high toxicity against Helicoverpa armigera and Spodoptera exigua. However, it lost insecticidal activities after being bathed in boiling water for 5 min. Acrystalliferous mutants of Bt9816C (Bt9816C-NP1 and Bt9816C-NP2) cured of its endogenous plasmids no longer possessed vip3A gene and toxicity. The 89 kD protein which existed in Bt9816C supernatant disappeared in the two mutants' supernatant; nevertheless, the two mutants still exhibited hemolytic and phospholipase C activity as Bt9816C did. The vip3A gene of Bt9816C, vip3Aa18, was cloned and expressed in Escherichia coli BL21. Bioassay demonstrated that the recombinant E. coli had high toxicity against S. exigua. Taken together, it suggested that Vip3A protein was responsible for the toxicity of Bt9816C culture supernatants.

Abstract: duced with a single species, Asaia bogorensis Yamada et al. 2000 in the family Acetobacteraceae Gillis and De Ley 1980 (Yamada et al., 2000). Since the second and the third species described were Asaia siamensis Katsura et al. 2001 and Asaia krungthepensis Yukphan et al. 2004, three species are in total reported (Katsura et al., 2001; Yamada et al., 2000; Yukphan et al., 2004c). The species assigned to the genus Asaia were characterized phenotypically by no oxidation or very weak oxidation of ethanol to acetic acid and by no growth in the presence of 0.35% (w/v) acetic acid. For the species-level identification and classification of acetic acid bacteria, including strains assigned to the genus Asaia, phenotypic characteristics such as acid production from sugars and sugar alcohols and assimilation of carbon compounds are generally utilized (Asai et al., 1964; Katsura et al., 2001; Lisdiyanti et al., 2002; Yamada et al., 1976, 1999, 2000; Yukphan et al., 2004c, d). However, data obtained by phenotypic characterization are not only difficult but also very often inaccurate. The phenotypic characteristics obtained are sometimes unreliable. On the other Identification of strains assigned to the genus Asaia Yamada et al. 2000 based on restriction analysis of 16S-23S rDNA internal transcribed spacer regions