記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE  

There are growing concerns about the impacts of neonicotinoid insecticides on ecosystems worldwide, and yet ecotoxicity of many of these chemicals at community or ecosystem levels have not been evaluated under realistic conditions. In this study, effects of two neonicotinoid insecticides, imidacloprid and dinotefuran, on aquatic insect assemblages were evaluated in experimental rice mesocosms. During the 5-month period of the rice-growing season, residual concentrations of imidacloprid were 5-10 times higher than those of dinotefuran in both soil and water. Imidadoprid treatment (10 kg/ha) reduced significantly the populations of Crocothemis servilia inariannae and Lyriothemis pachygastra nymphs, whereas those of Orthetrwn afbistylum speciosum increased slightly throughout the experimental period. However, Notonecta uiguttata, which numbers were high from the start, later declined, indicating possible delayed chronic toxicity, while Guignotus japonicus disappeared. In contrast, dinotefuran (10 kg/ha) did not decrease the populations of any species, but rather increased the abundance of some insects, particularly Chironominae spp. larvae and C. servllia mariannoe nymphs, with the latter being 1.7x higher than those of controls. This was an indirect effect resulting from increased prey (e.g., chironomid larvae) and lack of competition with other dragonfly species. The susceptibilities of dragonfly nymphs to neonicotinoids, particularly irnidadoprid, were consistent with those reported elsewhere. In general, imidacloprid had higher impacts on aquatic insects compared to dinotefuran.

DOI： 10.1016/j.ecoenv.2016.12.025

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC ANTIBACTERIAL & ANTIFUNGAL AGENTS, JAPAN  

Food is a basic necessity for human survival, but it is still the vehicle for the transmission of food borne disease. Various studies have examined the roles of spices, herbs, nuts, and semidried fruits, making the need for safe and convenient methods of decontamination a necessity. The current study determined the bacterial and fungal loads of 26 spices and herbs, 5 nuts, 10 semi-dried fruits and 5 other foods. Spices, herbs and semi-dried foods demonstrated the highest bacterial and fungal loads with the majority showing over 10(4) CFU/mL. Nuts and other foods showed growths ranging from 10(2) to 10(6) CFU/mL. The current study also attempted to determine the effects of heat and plasma treatment. The log reduction of bacterial growth after heat treatment (maximum: 120 min for 60 degrees C) was between 0.08 to 4.47, and the log reduction after plasma treatment (maximum: 40 min) ranged from 2.37 to 5.75. Spices showed the lowest rates of reduction, whereas the semi-dried and other foods showed moderate to high levels of decrease after heat treatment. The log reduction of fungal growth after heat treatment ranged from 0.27 to 4.40, and log reduction after plasma treatment ranged from 2.15 to 5.91. Furthermore, we validated the sterilization effect of plasma treatment against Bacillus spp. and Staphylococcus spp. by using scanning electron microscopy. Both treatment methods could prove to be advantageous in the agriculture related fields, enhancing the quality of the foods.

DOI： 10.4265/bio.21.243

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

Here, we report the draft genome sequence of Flammulina velutipes TR19, which was newly isolated from commercial strains in Japan. The genes related to fruiting body formation in the basidiomycete were identified by whole-genome analysis.

DOI： 10.1128/genomeA.00505-16

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD  

A new minor carotenoid (1) was isolated from the sea cucumber Plesiocolochirus minutus along with beta-carotene, echinenone, canthaxanthin, adonirubin, astaxanthin, and cucumariaxanthin A. The structure of 1 was determined to be 9Z,9'Z-(3R,SS,6S,3'R,5'S,6'S)-3,3'-dihydroxy-5,6,5',6'-tetrahydro-beta,beta-carotene4,4'-dione based on chemical and spectroscopic data. This compound was named 9Z,9'Z-tetrahydroastaxanthin. (C) 2015 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.tetlet.2015.09.060

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC ANTIBACTERIAL & ANTIFUNGAL AGENTS, JAPAN  

Bisphenol A (BPA, 2,2'-Bis(4-hydroxyphenyl)propane) is an artificial pollutant that is easily detected in soil and water environments. BPA decomposition and removal from the environment is relatively difficult due to its stability. This study evaluated the BPA decomposition and removal activities of the microbial community existing in the soil with or without Sphingomonas bisphenolicum AO1, and revealed the toxic effects of BPA towards the microbial community. The microbial community in soil was able to degrade BPA at 1.0 mg.g(-1) soil or lower, although its degradation was slow. On the other hand, BPA at more than 10 mg.g(-1) soil was not only degraded by the microbial community but also decreased its diversity, suggesting that BPA is harmful to many microorganisms. PCR-TTGE analysis and the cloned 16S rRNA gene sequence analysis indicated that Sphingomonadales, Xanthomonadales, Burkholderiales and Pseudomonadales in the microbial community might independently or cooperatively degrade BPA. On the other hand, supplementation with strain AO1 was able to significantly improve the BPA decomposition activity of the microbial community in soil even at 10 mg BPA.g(-1) soil, although BPA at 100 mg.g(-1) soil overwhelmed the BPA decomposition activity of strain AO1. Furthermore, it was also concluded that strain AO1 could not inhabit BPA purified soil after decomposition of BPA by strain AO1 and the soil microbial community, suggesting that the application of strain AO1 could be a low-burden method for the decomposition and removal of BPA from the natural environment.

DOI： 10.4265/bio.20.35

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Recombinant Corynebacterium glutamicum harboring genes for pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB) can produce ethanol under oxygen deprivation. We investigated the effects of elevating the expression levels of glycolytic genes, as well as pdc and adhB, on ethanol production. Overexpression of four glycolytic genes (pgi, pfkA, gapA, and pyk) in C. glutamicum significantly increased the rate of ethanol production. Overexpression of tpi, encoding triosephosphate isomerase, further enhanced productivity. Elevated expression of pdc and adhB increased ethanol yield, but not the rate of production. Fed-batch fermentation using an optimized strain resulted in ethanol production of 119 g/L from 245 g/L glucose with a yield of 95 % of the theoretical maximum. Further metabolic engineering, including integration of the genes for xylose and arabinose metabolism, enabled consumption of glucose, xylose, and arabinose, and ethanol production (83 g/L) at a yield of 90 %. This study demonstrated that C. glutamicum has significant potential for the production of cellulosic ethanol.

DOI： 10.1007/s00253-014-6223-4

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC ANTIBACTERIAL & ANTIFUNGAL AGENTS, JAPAN  

We have previously conducted a microflora analysis and examined the biofilm-forming activity of bacteria isolated from toilet bowl biofilms. In the present investigation, to reveal the strain involved in the formation of black dirt in toilet bowls, we performed a microflora analysis of the bacteria and fungi isolated from the black dirt of toilet bowls at ten homes. Among samples from different isolation sites and sampling seasons, although a similar tendency was not seen in bacterial microflora, Exophiala sp was detected in the fungal microflora from all samples of black dirt except for one, and constituted the major presence. By scanning electron microscope (SEM) analysis of the formed black dirt, SEM image at X 1,000 and X 5,000 magnification showed objects like hyphae and many bacteria adhering to them, respectively. Micro fourier transform infrared spectroscopy (micro FT-IR) and SEM with X-ray microanalysis (SEM-XMA) were used to investigate the components of black dirt. IR spectra of micro-FT-IA showed typical absorptions associated with amide compounds and protein, and the elements such as C, N, 0, Na, Mg, Al, Si, P, S, K, and Ba were detected with SEM-XMA. These results showed that black dirt had living body ingredients. Furthermore, Exophiala sp. and Cladosporium sp. strains, which were observed at a high frequency, accumulated 2-hydroxyjuglone (2-HJ) and flaviolin as one of the intermediates in the melanin biosynthetic pathway by the addition of a melanin synthesis inhibitor (tricyclazole) at the time of cultivation. These results suggested strongly that the pigment of black dirt in toilet bowls was melanin produced by Exophiala sp. and Cladosporium sp. strains.

DOI： 10.4265/bio.19.173

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC ANTIBACTERIAL & ANTIFUNGAL AGENTS, JAPAN  

To evaluate the sanitary conditions of toilets, the bacterial counts of the toilet bowl biofilms in 5 Kansai area and 11 Kansai and Kanto area homes in Japan were measured in winter and summer seasons, respectively. Isolates (128 strains) were identified by analyzing 16S ribosomal RNA sequences. The number of colonies and bacterial species from biofilms sampled in winter tended to be higher and lower, respectively, than those in summer. Moreover, the composition of bacterial communities in summer and winter samples differed considerably. In summer samples, biofilms in Kansai and Kanto areas were dominated by Blastomonas sp. and Mycobacterium sp., respectively. Methylobacterium sp. was detected in all toilet bowl biofilms except for one sample. Methylobacterium sp. constituted the major presence in biofilms along with Brevundimonas sp., Sphingomonas sp., and/or Pseudomonas sp. The composition ratio of the sum of their genera was 88.0 from 42.9% of the total bacterial flora. The biofilm formation abilities of 128 isolates were investigated, and results suggested that Methylobacterium sp. and Sphingomonas sp. were involved in biofilm formation in toilet bowls. The biofilm formation of a mixed bacteria system that included bacteria with the highest biofilm-forming ability in a winter sample was greater than mixture without such bacteria. This result suggests that isolates possessing a high biofilm-forming activity are involved in the biofilm formation in the actual toilet bowl. A bactericidal test against 25 strains indicated that the bactericidal activities of didecyldimethylammonium chloride (DDAC) tended to be higher than those of polyhexamethylene biguanide (PHMB) and N-benzyl-N,N-dimethyldodecylammonium chloride (ADBAC). In particular, DDAC showed high bactericidal activity against approximately 90% of tested strains under the 5 h treatment.

DOI： 10.4265/bio.18.129

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC ANTIBACTERIAL & ANTIFUNGAL AGENTS, JAPAN  

DDAC is an effective disinfectant used in the medical and food industries and the environmental field. However, skin irritation in humans occurs at high DDAC concentrations. In this study, we analyzed the combined effect of a low concentration (0.3 ppm) of DDAC and low concentrations (6, 8, and 10 ppm) of 37 products of nonionic surfactants on the bactericidal activity against S. aureus. No bactericidal activity was found at 0.3 ppm DDAC alone. Results showed that a combination of a low concentration of DDAC (0.3 ppm) and some nonionic surfactants tested (synergistic effect of five products >= 2.0) improved the bactericidal activity of DDAC. Synergistic effects of DDAC and some nonionic surfactants are desirable and were suggested to occur as follows. Test surfactants acted against the cell walls of S. aureus, which allowed DDAC to act easily on the lipid double membrane in the cell wall, thereby increasing the bactericidal activity of DDAC. In the present study, synergistic effects of a low concentration of DDAC and some nonionic surfactants were observed, a phenomenon that may be considerable value in future developments.

DOI： 10.4265/bio.17.175

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JAPAN OIL CHEMISTS SOC  

The biochemical properties of carotenoids from 2 species of freshwater bivalve, namely, (Unio douglasiae nipponensis and Anodonta lauta, and 2 species of freshwater snail, namely, Cipangopaludina chinensis laeta and Semisulcospira libertina, were investigated. Diatoxanthin and fucoxanthin were identified as major carotenoids in both bivalves. In contrast, lutein and zeaxanthin were found to be the major carotenoids in C. chinensis beta. In addition, a series of keto carotenoids was also identified in S. libertina.

DOI： 10.5650/jos.61.69

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MDPI AG  

Carotenoids of the corals Acropora japonica, A. secale, and A. hyacinthus, the tridacnid clam Tridacna squamosa, the crown-of-thorns starfish Acanthaster planci, and the small sea snail Drupella fragum were investigated. The corals and the tridacnid clam are filter feeders and are associated with symbiotic zooxanthellae. Peridinin and pyrrhoxanthin, which originated from symbiotic zooxanthellae, were found to be major carotenoids in corals and the tridacnid clam. The crown-of-thorns starfish and the sea snail D. fragum are carnivorous and mainly feed on corals. Peridinin-3-acyl esters were major carotenoids in the sea snail D. fragum. On the other hand, ketocarotenoids such as 7,8-didehydroastaxanthin and astaxanthin were major carotenoids in the crown-of-thorns starfish. Carotenoids found in these marine animals closely reflected not only their metabolism but also their food chains.

DOI： 10.3390/md9081419

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Here, focus is on Corynebacterium glutamicum mannose metabolic genes with the aim to improve this industrially important microorganism's ability to ferment mannose present in mixed sugar substrates. cgR_0857 encodes C. glutamicum's protein with 36% amino acid sequence identity to mannose 6-phosphate isomerase encoded by manA of Escherichia coli. Its deletion mutant did not grow on mannose and exhibited noticeably reduced growth on glucose as sole carbon sources. In effect, C. glutamicum manA is not only essential for growth on mannose but also important in glucose metabolism. A double deletion mutant of genes encoding glucose and fructose permeases (ptsG and ptsF, respectively) of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was not able to grow on mannose unlike the respective single deletion mutants with mannose utilization ability. A mutant deficient in ptsH, a general PTS gene, did not utilize mannose. These indicate that the glucose-PTS and fructose-PTS are responsible for mannose uptake in C. glutamicum. When cultured with a glucose and mannose mixture, mannose utilization of manA-overexpressing strain CRM1 was significantly higher than that of its wild-type counterpart, but with a strong preference for glucose. ptsF-overexpressing strain CRM2 co-utilized mannose and glucose, but at a total sugar consumption rate much lower than that of the wild-type strain and CRM1. Strain CRM3 overexpressing both manA and ptsF efficiently co-utilized mannose and glucose. Under oxygen-deprived conditions, high volumetric productivity of organic acids concomitant with the simultaneous consumption of the mixed sugars was achieved by the densely packed growth-arrested CRM3 cells.

DOI： 10.1007/s00253-010-3002-8

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：日本防菌防黴学会  

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：日本防菌防黴学会  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Wild-type Corynebacterium glutamicum produced 0.6 g l(-1) xylitol from xylose at a productivity of 0.01 g l(-1) h(-1) under oxygen deprivation. To increase this productivity, the pentose transporter gene (araE) from C. glutamicum ATCC31831 was integrated into the C. glutamicum R chromosome. Consequent disruption of its lactate dehydrogenase gene (ldhA), and expression of single-site mutant xylose reductase from Candida tenuis (CtXR (K274R)) resulted in recombinant C. glutamicum strain CtXR4 that produced 26.5 g l(-1) xylitol at 3.1 g l(-1) h(-1). To eliminate possible formation of toxic intracellular xylitol phosphate, genes encoding xylulokinase (XylB) and phosphoenolpyruvate-dependent fructose phosphotransferase (PTS(fru)) were disrupted to yield strain CtXR7. The productivity of strain CtXR7 increased 1.6-fold over that of strain CtXR4. A fed-batch 21-h CtXR7 culture in mineral salts medium under oxygen deprivation yielded 166 g l(-1) xylitol at 7.9 g l(-1) h(-1), representing the highest bacterial xylitol productivity reported to date.

DOI： 10.1007/s00253-009-2372-2

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC ANTIBACTERIAL & ANTIFUNGAL AGENTS, JAPAN  

One hundred and seven soil samples were collected from various places In Japan, and their bisphenol-A (BPA, 2,2-bis(4-Hydroxyphenyl)propane) degradative capacities were evaluated. Eighty-five soil samples possessed BPA degradative capacities, and 26 bacterial strains could be isolated as BPA-degrading bacterium. Sequence analysis of their 16S rRNA genes indicated that 22 isolates belonged to proteobacteria groups, and three of four Gram-positive bacterial strains, YA27, NO13, and NO15, were classified as Bacilli. All isolates except strain YA27 completely degraded 115 mu g/mL BPA in L medium but strain YA27 degraded only 50 mu g/mL BPA. Strain YA27 and three Sphingomonas sp. strains could also grow In basal salt media containing BPA as a sole carbon source (BSMB medium). In HPLC analyses, some isolates, including the three Sphingomonas strains, produced some BPA metabolites in their cultures although the others, Including strain YA27, produced no detectable metabolite. Furthermore, the Pseudomonas strains SU1 and SU4 produced some BPA metabolites that were different from the metabolites detected in the degradation of BPA by the S. bisphenolicum strain AO1. These results suggested that all Isolates could be applicable to the bioremediation of BPA-polluted soil and water. Furthermore, we suggest that Bacillus sp. YA27 and Pseudomonas SU1 and SU4 may exhibit novel BPA metabolism pathways that are distinct from that of S. bisphenolicum AO1.

DOI： 10.4265/bio.14.161

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Corynebacterium glutamicum strains CRA1 and CRX2 are able to grow on l-arabinose and d-xylose, respectively, as sole carbon sources. Nevertheless, they exhibit the major shortcoming that their sugar consumption appreciably declines at lower concentrations of these substrates. To address this, the C. glutamicum ATCC31831 l-arabinose transporter gene, araE, was independently integrated into both strains. Unlike its parental strain, resultant CRA1-araE was able to aerobically grow at low (3.6 g center dot l(-1)) l-arabinose concentrations. Interestingly, strain CRX2-araE grew 2.9-fold faster than parental CRX2 at low (3.6 g center dot l(-1)) d-xylose concentrations. The corresponding substrate consumption rates of CRA1-araE and CRX2-araE under oxygen-deprived conditions were 2.8- and 2.7-fold, respectively, higher than those of their respective parental strains. Moreover, CRA1-araE and CRX2-araE utilized their respective substrates simultaneously with d-glucose under both aerobic and oxygen-deprived conditions. Based on these observations, a platform strain, ACX-araE, for C. glutamicum-based mixed sugar utilization was designed. It harbored araBAD for l-arabinose metabolism, xylAB for d-xylose metabolism, d-cellobiose permease-encoding bglF (317A) , beta-glucosidase-encoding bglA and araE in its chromosomal DNA. In mineral medium containing a sugar mixture of d-glucose, d-xylose, l-arabinose, and d-cellobiose under oxygen-deprived conditions, strain ACX-araE simultaneously and completely consumed all sugars.

DOI： 10.1007/s00253-009-2065-x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER SOC MICROBIOLOGY  

Corynebacterium glutamicum ATCC 31831 grew on L-arabinose as the sole carbon source at a specific growth rate that was twice that on D-glucose. The gene cluster responsible for L-arabinose utilization comprised a six-cistron transcriptional unit with a total length of 7.8 kb. Three L-arabinose-catabolizing genes, araA (encoding L-arabinose isomerase), araB (L-ribulokinase), and araD (L-ribulose-5-phosphate4-epimerase), comprised the araBDA operon, upstream of which three other genes, araR (LacI-type transcriptional regulator), araE (L-arabinose transporter), and galM ( putative aldose 1-epimerase), were present in the opposite direction. Inactivation of the araA, araB, or araD gene eliminated growth on L-arabinose, and each of the gene products was functionally homologous to its Escherichia coli counterpart. Moreover, compared to the wild-type strain, an araE disruptant exhibited a > 80% decrease in the growth rate at a lower concentration of L-arabinose (3.6 g liter(-1)) but not at a higher concentration of L-arabinose ( 40 g liter(-1)). The expression of the araBDA operon and the araE gene was L-arabinose inducible and negatively regulated by the transcriptional regulator AraR. Disruption of araR eliminated the repression in the absence of L-arabinose. Expression of the regulon was not repressed by D-glucose, and simultaneous utilization of L-arabinose and D-glucose was observed in aerobically growing wild-type and araR deletion mutant cells. The regulatory mechanism of the L-arabinose regulon is, therefore, distinct from the carbon catabolite repression mechanism in other bacteria.

DOI： 10.1128/AEM.02912-08

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Corynebacterium glutamicum R was metabolically engineered to broaden its sugar utilization range to D-xylose and D-cellobiose contained in lignocellulose hydrolysates. The resultant recombinants expressed Escherichia coli xylA and xylB genes, encoding D-xylose isomerase and xylulokinase, respectively, for D-xylose utilization and expressed C. glutamicum R bglF(317A) and bglA genes, encoding phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) beta-glucoside-specific enzyme IIBCA component and phospho-beta-glucosidase, respectively, for D-cellobiose utilization. The genes were fused to the non-essential genomic regions distributed around the C. glutamicum R chromosome and were under the control of their respective constitutive promoter trc and tac that permitted their expression even in the presence of D-glucose. The enzyme activities of resulting recombinants increased with the increase in the number of respective integrated genes. Maximal sugar utilization was realized with strain X5C1 harboring five xylA-xylB clusters and one bglF(317A)-bglA cluster. In both D-cellobiose and D-xylose utilization, the sugar consumption rates by genomic DNA-integrated strain were faster than those by plasmid-bearing strain, respectively. In mineral medium containing 40 g l(-1) D-glucose, 20 g l(-1) D-xylose, and 10 g l(-1) D-cellobiose, strain X5C1 simultaneously and completely consumed these sugars within 12 h and produced predominantly lactic and succinic acids under growth-arrested conditions.

DOI： 10.1007/s00253-008-1703-z

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL  

Aims: To clone and characterize the genes bisdA and bisdB, encoding Ferredoxin(bisd) (Fd(bisd)) and cytochrome P450(bisd) (P450(bisd)), respectively, from the bisphenol A (BPA) degrading Sphingomonas bisphenolicum strain AO1.
Methods and Results: The 3.7 kb region containing bisdA and bisdB was cloned by genome walking and colony hybridization. The deduced N-terminal amino acid sequences of bisdA and bisdB were consistent with those of Fd(bisd) and P450(bisd) proteins characterized in our previous report. Two transposase genes, tnpA1 and tnpA2, were also located upstream and downstream of bisdAB. From amino acid sequence analysis, P450(bisd) has two conserved regions corresponding to the oxygen and heme binding regions of the bacterial cytochrome P450 family. Fd(bisd) was similar to putidaredoxin-type [2Fe-2S] ferredoxins. Escherichia coli BL21 (DE3) cells bearing bisdB- and bisdAB-recombinant pET19b were able to degrade BPA. A spontaneous mutant, strain AO1L, which was unable to degrade BPA, was isolated from the stock culture, and it was confirmed that strain AO1L had no bisdAB region.
Conclusions: P450(bisd) monooxygenase sytem, encoded by bisdAB, is one system required for BPA hydroxylation in S. bisphenolicum strain AO1.
Significance and Impact of the Study: Our results indicate that bisdAB are key genes for BPA degradation in S. bisphenolicum strain AO1.

DOI： 10.1111/j.1365-2672.2008.03843.x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Corynebacterium glutamicum was metabolically engineered to broaden its substrate utilization range to include the pentose sugar L-arabinose, a product of the degradation of lignocellulosic biomass. The resultant CRA1 recombinant strain expressed the Escherichia coli genes araA, araB, and araD encoding L-arabinose isomerase, L-ribulokinase, and L-ribulose-5-phosphate 4-epimerase, respectively, under the control of a constitutive promoter. Unlike the wild-type strain, CRA1 was able to grow on mineral salts medium containing L-arabinose as the sole carbon and energy source. The three cloned genes were expressed to the same levels whether cells were cultured in the presence of D-glucose or L-arabinose. Under oxygen deprivation and with L-arabinose as the sole carbon and energy source, strain CRA1 carbon flow was redirected to produce up to 40, 37, and 11%, respectively, of the theoretical yields of succinic, lactic, and acetic acids. Using a sugar mixture containing 5% D-glucose and 1% L-arabinose under oxygen deprivation, CRA1 cells metabolized L-arabinose at a constant rate, resulting in combined organic acids yield based on the amount of sugar mixture consumed after D-glucose depletion (83%) that was comparable to that before D-glucose depletion (89%). Strain CRA1 is, therefore, able to utilize L-arabinose as a substrate for organic acid production even in the presence of D-glucose.

DOI： 10.1007/s00253-007-1244-x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER SOC MICROBIOLOGY  

In a previous study (M. Sasaki, J. Maki, K. Oshiman, Y. Matsumura, and T. Tsuchido, Biodegradation 16:449-459, 2005), the cytochrome P450 monooxygenase system was shown to be involved in bisphenol A (BPA) degradation by Sphingomonas sp. strain AO1. In the present investigation, we purified the components of this monooxygenase, cytochrome P450 (P450(bisd)), ferredoxin (Fd(bisd)), and ferredoxin reductase (Red(bisd)). We demonstrated that P450(bisd) and Fd(bisd) are homodimeric proteins with molecular masses of 102.3 and 19.1 kDa, respectively, by gel filtration chromatography analysis. Spectroscopic analysis of Fd(bisd) revealed the presence of a putidaredoxin-type [2Fe-2S] cluster. P450(bisd), in the presence of Fd(bisd), Red(bisd), and NADH, was able to convert BPA. The K-m and k(cat) values for BPA degradation were 85 +/- 4.7 mu M and 3.9 +/- 0.04 min(-1), respectively. NADPH, spinach ferredoxin, and spinach ferredoxin reductase resulted in weak monooxygenase activity. These results indicated that the electron transport system of P450(bisd) might exhibit strict specificity. Two BPA degradation products of the P450(bisd) system were detected by high-performance liquid chromatography analysis and were thought to be 1,2-bis(4-hydroxyphenyl)-2-propanol and 2,2-bis(4-hydroxyphenyl)-1-propanol based on mass spectrometry-mass spectrometry analysis. This is the first report demonstrating that the cytochrome P450 monooxygenase system in bacteria is involved in BPA degradation.

DOI： 10.1128/AEM.71.12.8024-8030.2005

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PubMed

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

The capacity and pathway of bisphenol A [BPA; 2,2- bis(4-hydroxyphenyl) propane] degradation in Sphingomonas sp. strain AO1, which was isolated from the soil of a vegetable-growing field in Japan, were investigated. The bacterial strain was able to grow in a basal mineral salt medium containing BPA as the sole carbon source ( BSMB medium), and was able to degrade 115 mug ml(-1) BPA in 6 h in L medium. Several BPA metabolites were detected in the culture supernatant by HPLC and then identified by GC-MS and LC-MS-MS. These compounds were confirmed to be the same as those reported for other BPA-degrading bacteria. BPA degradation by cells in the basal mineral salt medium was induced by BPA, and activity was detected only in the intracellular soluble fraction in the presence of coenzymes, such as NADH, NAD+, NADPH or NADP+. The addition of metyrapone, a cytochrome P450 inhibitor, to BSMB medium resulted in a decrease in BPA degradation and cell growth. The BPA-degradation activity of the intracellular soluble fraction was also inhibited by the cytochrome P450 inhibitor. Carbon monoxide difference spectra indicated that cytochrome P450 was present in the cells and that the amount of cytochrome P450 corresponded to the cellular BPA-degradation activity. Our results provide evidence that the cytochrome P450 system is involved in BPA metabolism in Sphingomonas sp. strain AO1.

DOI： 10.1007/s10532-004-5023-4

Web of Science

PubMed

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

We purified and characterized a 39-kDa Bacillus subtilis 168 nuclease that has been suggested in this laboratory to be involved in chromosomal DNA degradation induced by lethal heat and cold shock treatments in vivo. The nuclease activity was inhibited in vitro by aurintricalboxylic acid but not by Zn2+. By the mutant analysis, we identified the 39-kDa nuclease as a product of yokF gene. The yokF gene contained a putative lipoprotein signal peptide motif. After in vivo exposure to lethal heat and cold stresses, the chromosomal DNA fragmentation was reduced in the yokF mutant, which demonstrated about a 2-10-fold higher survival rate than the wild type. The yokF mutant was found to be more sensitive to mitomycin C than the wild type. The transformation efficiency of the yokF mutant was about 10 times higher than that of the wild type. It is suggested that when B. subtilis cells are exposed to a stressful thermal shock resulting in membrane perturbation, YokF nuclease consequently dislocates into the cytoplasm and then attacks DNA.

DOI： 10.1074/jbc.M106205200

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記述言語：日本語   出版者・発行元：日本防菌防黴学会  

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