• Title/Summary/Keyword: ${\beta}$-proteobacterium

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Purification and Properties of Chitosanase from Chitinolytic $\beta$-Proteobacterium KNU3

  • Yi, Jae-Hyoung;Jang, Hong-Ki;Lee, Sang-Jae;Lee, Keun-Eok;Choi, Shin-Geon
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.337-343
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    • 2004
  • A bacterial strain concurrently producing extracellular chitosanase and chitinase was isolated from soil and identified as a member of the $\beta$-subgroup of Proteobacteria through its 16S rRNA analysis and some biochemical analyses. The newly discovered strain, named as KNU3, had 99% homology of its 16S rRNA sequence with chitinolytic $\beta$-Proteobacterium CTE108. Strain KNU3 produced 34 kDa of chitosanase in addition to two chitinases of 68 kDa and 30 kDa, respectively. The purified chitosanase protein (ChoK) showed activity toward soluble, colloidal, and glycol chitosan, but did not exhibit any activity toward colloidal chitin. The optimum pH and temperature of ChoK were 6.0 and $70^{\circ}C$, respectively. The chitosanase was stable in the pH 4.0 to 8.0 range at $70^{\circ}C$, while enzyme activity was relatively stable at below $45^{\circ}C$. MALDI-TOF MS and N-terminal amino acid sequence analyses indicated that ChoK protein is related to chitosanases from Matsuebacter sp. and Sphingobacterium multivorum. HPLC analysis of chitosan lysates revealed that glucosamine tetramers and hexamers were the major products of hydrolysis.

Cloning and Expression of a Novel Chitosanase Gene (choK) from $\beta$-Proteobacterium KNU3 by Double Inverse PCR

  • Yi, Jae-Hyoung;Lee, Keun-Eok;Choi, Shin-Geon
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.563-569
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    • 2004
  • The DNA sequence of the chitosanase gene (choK) from $\beta$-Proteobacterium KNU3 showed an 1,158-bp open reading frame that encodes a protein of 386 amino acids with a novel 74 signal peptide. The degenerated primers based on the partial deduced amino acid sequences from MALDI- TOF MS analyses yielded the 820 bp of the PCR product. Based on this information, double inverse PCR cloning experiments, which use the two specific sets of PCR primers rather than single set primers, identified the unknown 1.2 kb of the choK gene. Subsequently, a 1.8 kb of full choK gene was cloned from another PCR cloning experiment and it was then subcloned into pGEM T-easy and pUC18 vectors. The recombinant E. coli clone harboring recombinant pUC18 vector produced a clear halo around the colony in the glycol chitosan plates. The recombinant ChoK protein was secreted into medium in a mature form while the intracellular ChoK was produced without signal peptide cleavage. The activity staining of PAGE showed that the recombinant ChoK protein was identical to the chitosanase of wild-type. The comparison of deduced amino acid sequences of choK revealed that there is 92% identity with that of Sphingobacterium multivorum chitosanase. Judging from the conserved module in other bacterial chitosanases, chitosanase of KNU3 strain (ChoK) belongs to the family 80 of glycoside hydrolases.

Characterization of a Xanthorhodopsin-homologue from the North Pole

  • Kim, Se Hwan;Cho, Jang-Cheon;Jung, Kwang-Hwan
    • Rapid Communication in Photoscience
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    • v.2 no.2
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    • pp.60-63
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    • 2013
  • Rhodopsins belong to a family of membrane-embedded photoactive retinylidene proteins. One opsin gene was isolated from ${\beta}$-proteobacterium (IMCC9480) which had been collected at the North Pole. It is very similar to Xanthorhodopin (XR) of HTCC2181. In this study, we carried out basic characterization of the rhodopsin. It has ${\lambda}max$ of 536, 554, and 546 nm at pH 4.0, 7.0, and 10.0, respectively. Since the pKa of its proton acceptor is around 6.27, we measured its proton pumping activity and photocycling rate at pH 8.0. It has a typical proton acceptor (D99) and donor (E110) which mediate proton translocation from intracellular to extracellular region when deduced from the sequence alignments. On the basis of in vitro proton pumping activity, it was proposed to have fast photocycling rate with M and O intermediates, indicating that it is a typical ion-pumping rhodopsin. Since the XR has not yet been expressed in any other heterologous expression system, we tried to get much more information about the XR through the XR-homologue rhodopsin.

Evaluation of Microbes through Microfiltration within the Water Treatment Processes (정밀여과막 및 입상활성탄을 이용한 수처리 공정에 따른 박테리아 거동 평가)

  • Shim, Moon Jung;Lim, Jae Won;Kim, Tae Ue
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.230-236
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    • 2016
  • Economic growth has increased the living standards around the world. Water pollution, in particular, is a public relations issue because it poses a direct threat to everyone's lives. As of recently, the production of taste and odor (T&O) compounds has been a common problem in the water industry. The adsorption process using granular activated carbon (GAC) has been the most widely used process. The objectives of this study were to evaluate the microorganisms before and after the backwashing of GAC and to identify the species of the microorganisms found. Five dominants microorganisms were confirmed after the microfiltration process from backwashing of GAC, and the dominant bacterial species were found to be ${\beta}$-proteobacterium species, Porphyrobacter donghaensis, Polaromonas rhizophaerae, Hydrogenophaga species, and Pseudonocardia species. However, when UV treatment after microfiltration was performed, Hydrogenophaga species and Psedonocardia species were eliminated. Herein, I conclude that the UV treatment post microfiltration process is more efficient than microfiltration process alone. The findings of this study may provide useful information regarding the management of microfiltration process.

Isolation and Characterization of Extracellular Polymeric Substances (EPS)-producing bacteria for restoration of burnt forest soils (산불토양복원을 위한 Extracellular Polymeric Substances (EPS) 생성세균의 분리, 동정 및 특성에 관한 연구)

  • Lee, Gun-Young;Song, In-Geun;Chung, Jae-Chun;Kim, Young-Jun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.12 no.4
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    • pp.139-147
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    • 2004
  • We have isolated two bacterial strains, FM-02 and AL-02, which produced EPS from forest soil for the restoration of forest fire by promoting soil aggregation. FM-02 was found to be Gram negative rod and belong to Beta Proteobacterium sp. through 16s-rDNA sequence analysis, and AL-02 was Gram positive rod and showed 81% of similarity to Zoogloea sp. through the analysis of 16s-rDNA sequence. FM-02 and AL-02 produced about 1.8g and 8.3g of EPS, respectively, per 1L of culture as dry weight. Flocculation activity (FA) was also measured in two strains. FM-02 showed 2.31 FA against active carbon, and AL-02 showed 6.21 FA against kaolin clay. From these results, we expect that AL-02 strain will be applied as a good biological material for the reduction of forest soil erosion by wild and rain after fire through promoting coagulation of soil particles.

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Monitoring of Bacteria using PCR Method and Inactivation with Ozone and UV (PCR법에 의한 박테리아의 동정 및 오존과 UV에 의한 제거 연구)

  • Oh, Byung-Soo;Ju, Seul;Kim, Kyoung-Suk;Kang, Tae-Hee;Lee, Ji-Young;Lee, Hye-Young;Kang, Joon-Wun
    • Journal of Korean Society on Water Environment
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    • v.20 no.2
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    • pp.170-175
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    • 2004
  • The purpose of this study was to monitor bacteria present in raw water and to investigate the effect of ozone, UV and combined ozone/UV processes for inactivating bacteria. Both polymerase chain reaction (PCR)-fragment length polymorphic analysis (PRA) and PCR-sequence analysis (PSA) were applied for the simultaneous analysis of numerous bacteria species present in each tested water, such as drinking water (DRW), drinking water source (DRWS) and sewage effluent water (SEW). According to the result, the number of detected bacteria species was zero in DRW, 58 in DRWS and 13 in SEW. After treatment of the each process, the ozone/UV process was the most successful for inactivating almost all bacteria. However, it was found that Flavobacterium sp., Pseudomonas sp. and Beta proteobacterium sp. had strong resistant to all tested processes, requiring further detailed study.

Ramlibacter ginsenosidimutans sp. nov., with Ginsenoside-Converting Activity

  • Wang, Liang;An, Dong-Shan;Kim, Song-Gun;Jin, Feng-Xie;Kim, Sun-Chang;Lee, Sung-Taik;Im, Wan-Taek
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.311-315
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    • 2012
  • A novel ${\beta}$-proteobacterium, designated BXN5-$27^T$, was isolated from soil of a ginseng field of Baekdu Mountain in China, and was characterized using a polyphasic approach. The strain was Gram-staining-negative, aerobic, motile, non-spore-forming, and rod shaped. Strain BXN5-$27^T$ exhibited ${\beta}$-glucosidase activity that was responsible for its ability to transform ginsenoside $Rb_1$ (one of the dominant active components of ginseng) to compound Rd. Phylogenetic analysis based on 16S rRNA gene sequences showed that this strain belonged to the family Comamonadaceae; it was most closely related to Ramlibacter henchirensis $TMB834^T$ and Ramlibacter tataouinensis$TTB310^T$ (96.4% and 96.3% similarity, respectively). The G+C content of the genomic DNA was 68.1%. The major menaquinone was Q-8. The major fatty acids were $C_{16:0}$, summed feature 4 (comprising $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2OH), and $C_{17:0}$ cyclo. Genomic and chemotaxonomic data supported the affiliation of strain BXN5-$27^T$ to the genus Ramlibacter. However, physiological and biochemical tests differentiated it phenotypically from the other established species of Ramlibacter. Therefore, the isolate represents a novel species, for which the name Ramlibacter ginsenosidimutans sp. nov. is proposed, with the type strain being BXN5-$27^T$ (=DSM $23480^T$ = LMG $24525^T$ = KCTC $22276^T$).

Effect of Feeding Bacillus subtilis natto on Hindgut Fermentation and Microbiota of Holstein Dairy Cows

  • Song, D.J.;Kang, H.Y.;Wang, J.Q.;Peng, H.;Bu, D.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.4
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    • pp.495-502
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    • 2014
  • The effect of Bacillus subtilis natto on hindgut fermentation and microbiota of early lactation Holstein dairy cows was investigated in this study. Thirty-six Holstein dairy cows in early lactation were randomly allocated to three groups: no B. subtilis natto as the control group, B. subtilis natto with $0.5{\times}10^{11}cfu$ as DMF1 group and B. subtilis natto with $1.0{\times}10^{11}cfu$ as DMF2 group. After 14 days of adaptation period, the formal experiment was started and lasted for 63 days. Fecal samples were collected directly from the rectum of each animal on the morning at the end of eighth week and placed into sterile plastic bags. The pH, $NH_3$-N and VFA concentration were determined and fecal bacteria DNA was extracted and analyzed by DGGE. The results showed that the addition of B. subtilus natto at either treatment level resulted in a decrease in fecal $NH_3$-N concentration but had no effect on fecal pH and VFA. The DGGE profile revealed that B. subtilis natto affected the population of fecal bacteria. The diversity index of Shannon-Wiener in DFM1 decreased significantly compared to the control. Fecal Alistipes sp., Clostridium sp., Roseospira sp., beta proteobacterium were decreased and Bifidobacterium was increased after supplementing with B. subtilis natto. This study demonstrated that B. subtilis natto had a tendency to change fecal microbiota balance.