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Non-ureolytic calcium carbonate precipitation by Lysinibacillus sp. YS11 isolated from the rhizosphere of Miscanthus sacchariflorus
Yun Suk Lee , Hyun Jung Kim , Woojun Park
J. Microbiol. 2017;55(6):440-447.   Published online May 28, 2017
DOI: https://doi.org/10.1007/s12275-017-7086-z
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AbstractAbstract
Although microbially induced calcium carbonate precipita-tion (MICP) through ureolysis has been widely studied in en-vironmental engineering fields, urea utilization might cause environmental problems as a result of ammonia and nitrate production. In this study, many non-ureolytic calcium car-bonate-precipitating bacteria that induced an alkaline envi-ronment were isolated from the rhizosphere of Miscanthus sacchariflorus near an artificial stream and their ability to pre-cipitate calcium carbonate minerals with the absence of urea was investigated. MICP was observed using a phase-contrast microscope and ion-selective electrode. Only Lysinibacillus sp. YS11 showed MICP in aerobic conditions. Energy disper-sive X-ray spectrometry and X-ray diffraction confirmed the presence of calcium carbonate. Field emission scanning elec-tron microscopy analysis indicated the formation of morpho-logically distinct minerals around cells under these condi-tions. Monitoring of bacterial growth, pH changes, and Ca2+ concentrations under aerobic, hypoxia, and anaerobic con-ditions suggested that strain YS11 could induce alkaline con-ditions up to a pH of 8.9 and utilize 95% of free Ca2+ only under aerobic conditions. Unusual Ca2+ binding and its re-lease from cells were observed under hypoxia conditions. Bio-film and extracellular polymeric substances (EPS) formation were enhanced during MICP. Strain YS11 has resistance at high pH and in high salt concentrations, as well as its spore- forming ability, which supports its potential application for self-healing concrete.

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Research Support, Non-U.S. Gov'ts
Analysis of the Abilities of Endophytic Bacteria Associated with Banana Tree Roots to Promote Plant Growth
Leandro Fernandes Andrade , Gleika Larisse Oliveira Dorasio de Souza , Silvia Nietsche , Adelica Aparecida Xavier , Marcia Regina Costa , Acleide Maria Santos Cardoso , Marlon Cristian Toledo Pereira , Débora Francine Gomes Silva Pereira
J. Microbiol. 2014;52(1):27-34.   Published online January 4, 2014
DOI: https://doi.org/10.1007/s12275-014-3019-2
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  • 55 Crossref
AbstractAbstract
A total of 40 endophytic bacterial isolates obtained from banana tree roots were characterized for their biotechnological potential for promoting banana tree growth. All isolates had at least one positive feature. Twenty isolates were likely diazotrophs and formed pellicles in nitrogen-free culture medium, and 67% of these isolates belonged to the genus Bacillus sp. The isolates EB-04, EB-169, EB-64, and EB-144 had N fixation abilities as measured by the Kjeldahl method and by an acetylene reduction activity assay. Among the 40 isolates, 37.5% were capable of solubilizing inorganic phosphate and the isolates EB-47 and EB-64 showed the highest solubilization capacity. The isolate EB-53 (Lysinibacillus sp.) had a high solubilization index, whereas 73% of the isolates had low solubilization indices. The synthesis of indole-3- acetic acid (IAA) in the presence of L-tryptophan was detected in 40% of the isolates. The isolate EB-40 (Bacillus sp.) produced the highest amount of IAA (47.88 μg/ml) in medium supplemented with L-tryptophan and was able to synthesize IAA in the absence of L-tryptophan. The isolates EB-126 (Bacillus subtilis) and EB-47 (Bacillus sp.) were able to simultaneously fix nitrogen, solubilize phosphate and produce IAA in vitro. The results of this study demonstrated that the isolates analyzed here had diverse abilities and all have the potential to be used as growth-promoting microbial inoculants for banana trees.

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Isolation and Characterization of Plant Growth-Promoting Rhizobacteria from Wheat Roots by Wheat Germ Agglutinin Labeled with Fluorescein Isothiocyanate
Jian Zhang , Jingyang Liu , Liyuan Meng , Zhongyou Ma , Xinyun Tang , Yuanyuan Cao , Leni Sun
J. Microbiol. 2012;50(2):191-198.   Published online April 27, 2012
DOI: https://doi.org/10.1007/s12275-012-1472-3
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AbstractAbstract
Thirty-two isolates were obtained from wheat rhizosphere by wheat germ agglutinin (WGA) labeled with fluorescein isothiocyanate (FITC). Most isolates were able to produce indole acetic acid (65.6%) and siderophores (59.3%), as well as exhibited phosphate solubilization (96.8%). Fourteen isolates displayed three plant growth-promoting traits. Among these strains, two phosphate-dissolving ones, WS29 and WS31, were evaluated for their beneficial effects on the early growth of wheat (Triticum aestivum Wan33). Strain WS29 and WS31 significantly promoted the development of lateral roots by 34.9% and 27.6%, as well as increased the root dry weight by 25.0% and 25.6%, respectively, compared to those of the control. Based on 16S rRNA gene sequence comparisons and phylogenetic positions, both isolates were determined to belong to the genus Bacillus. The proportion of isolates showing the properties of plant growth-promoting rhizobacteria (PGPR) was higher than in previous reports. The efficiency of the isolation of PGPR strains was also greatly increased by WGA labeled with FITC. The present study indicated that WGA could be used as an effective tool for isolating PGPR strains with high affinity to host plants from wheat roots. The proposed approach could facilitate research on biofertilizers or biocontrol agents.

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Effect of copper on the growth and methanol dehydrogenase activity of methylobacillusd sp. strain SK1 DSM 8269
Kim, Si W. , Kim, Young M.
J. Microbiol. 1996;34(2):172-178.
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AbstractAbstract
Methylobacillus sp. strain SK1, which grows only on methanol, was found to grow in the absence of added copper. The doubling time (t_d = 1.3 h) of the bacterium growing at the exponential growth phase at 30℃ in the absence of copper was the same as that of the cell growing in the presence of copper. The bacterium growing after the exponential phase in the absence of copper, however, grew faster than the cell growing in the presence of copper. Cells harvested after thee early stationary phase in the presence of copper were found to exhibit no methanol dehydrogenase (MDH) activity, but the amount and subunit structure of the enzyme in the cells were almost the same as that in cells harboring active MDH. Pellets of the cells harvested after the early stationary phase in the presence of copper were pale green. Cell-free extracts prepared from cells harvested at the early stationary phase in the presence of copper were pink and exhibited MDH activity, but it turned dark-green rapidly from the surface under air. The green-colored portions of the extracts showed no MDH activity and contained c-type cytochromes that were oxidized completely. The inactive MDH activity and contained c-type cytochromes that were oxidized completely. The inactive MDH proteins in the green portions were found to have antigenic sites identical to those of the active one as the inactive MDHs in cells grown in the presence of copper. The bacterium was found to accumulate copper actively during the exponential growth phase. MDH prepared from cells grown in the presence or absence of copper was found to be more stable under nitrogen gas than under air. Methanol at 10 mM was found to enhance the stability of the MDH under air.
Isolation and characterization of Bacillus sp. KD1014 producing carboxymethyl-cellulase
Lee, Kyung Dong , Kim, Jung Ho , Kim Hoon
J. Microbiol. 1996;34(4):305-310.
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AbstractAbstract
A microorganism producing carboxymethyl-cellulase (CMCase) was isoalted from 300 soil and compost samples. The isolated was identified as Bacillus sp. by Biolog^TM test and fatty acid analysis, and named as Bacillus sp. KD1014. The isolate could degrade, in addition to CMC, various kinds of polysaccharides such as levan, xylan, starch, and filter paper but hardly degrade microcrystaline Avicel. The optimum growth and CMCase production of the isolate was observed between 16-and 25 hr-culture at 45℃ and pH 5.0. The maximum CMCase activity was observed at pH 4.5 and 60℃. The CMCase was found to bind to Avicel. The CMCase was internally cleaved as growth continued. When crude supernatant was used for activity staining, three major bands were detected on a native gel, however, only on major band was detected on a denaturating gel after removal of the detergent.
A possible mechanism responsible for translocation and secretion an alkaliphilic bacillus sp. S-1 pullulanase
Shim, Jae Kyoung , Kim, Kyoung Sook , Kim, Cheorl Ho
J. Microbiol. 1997;35(3):213-221.
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AbstractAbstract
The secretion of the alkaliphilic Bacillus sp. S-1 extracellular pullulanase involves translocation across the cytoplasmic membrane of the Gram-positive bacterial cell envelope. Translocation of the intracellular pullulanase PUL-I, was traced to elucidate the mechanism and pathway of protein secretion from an alkaliphilic Bacillus sp. S-1. Pullulanase could be slowly but quantitatively released into the medium during growth of the cells in medium containing proteinase K. The released pullulanase lacked the N-terminal domain. The N-terminus is the sole membrane anchor in the pullulanase protein and was not affected by proteases, confirming that it is not exposed on the cell surface. Processing of a 180,000M_r pullulanase to a 140,000M_r polypeptide has been demonstrated in cell extracts using antibodies raised against 140,000M_r extracellular form. Processing of the 180,000 M_r protein occured during the preparation of extracts in an alkaline pH condition. A modified rapid extraction procedure suggested that the processing event also occured in vivo. Processing apparently increased the activity of pullulanase. The western blotting analysis with mouse anti-serum against 140-kDa extracellular pullulanase PUL-E showed that PUL-I is processed into PUL-X via intermediate form of PUL-E. Possible explanationa for the translocation are discussed.

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