• Korean Journal of Microbiology
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• v.24 no.3
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• pp.228-235
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• 1986

The aims of the present studies were to develop conditions for the spheroplast formation of Zymomonas mobilis and regeneration of the spheroplasts to normal cells in synthetic media. Z. mobilis cells harvested from exponential growth phase were treated with lysozyme, mutanolysin, and glycine in various conditions. It was found that spheroplasts were formed only with the treatment of glycine but not with the enzymes treatments. It was therefore considered that the tetrapeptide strand of peptide strand of peptidoglycan might play more important roles than the glycan strand in maintaining the vital mechanical function of Z. mobilis. It was found also that removal of outher membrane was the major problem in protoplast formation of Z.mobilis. As results, It was observed that over 85% of cells were readly converted to spheroplasts with sole glycine treatment for 4 hr and 7-10% of the spheroplasts were regenerated to normal cells in synthetic media.

• Journal of Microbiology
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• v.33 no.2
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• pp.126-127
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• 1995

A Zymomonas mobilis DNA fragment consisting of 207 nucleotides, which corresponded to the 5'-flanking region of an adhB gene encoding alcohol dehydrogenase II, was fused to the structural gene coding for a Bacillus endo-.betha.--1, 4-glucanase. The Z. mobilis DNA framgment waw identified to promote 50-fold increase in the expression of endo-.betha.1. 4 glucanase gene in Escherichia coli.

• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.309-315
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• 1998

An extracellular levansucrase, which catalyzes the formation of levan from sucrose, from the culture broth of Zymomonas mobilis ZM1 was purified by conventional column purification methods. The final purification yield was 18.3 fold of the crude enzyme from Z. mobilis, with 16.5 % of the enzyme recovered in the preparation step. The molecular weight of the enzyme was estimated to be 91,000 by Superose 12 gel filtration, and 45,000 by SDS-PAGE, indicating that levansucrase is a dimer. The optimum pH for the enzyme activity was around pH 4.0 for sucrose hydrolysis, and was around pH 5.0 for levan formation. The enzyme was inhibited by some metal ions, such as Hg$\^$2+/ and Cu2$\^$2+/, and 50% of inhibition was observed with 5mM EDTA. The enzyme activity was enhanced by the presence of detergent Triton X-100, but inhibited by SDS completely The enzyme catalyzes the liberation of reducing sugars, oligosacccharides and the formation of fructose polymer(levan). The enzyme also catalyzes the transfructosylation reaction of fructose moiety from sucrose to various sugar acceptor molecules, including sugar alcohols.

• Journal of Microbiology and Biotechnology
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• v.19 no.7
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• pp.666-674
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• 2009

An electrochemical bioreactor (ECB) composed of a cathode compartment and an air anode was used in this study to characterize the ethanol fermentation of Zymomonas mobilis. The cathode and air anode were constructed of modified graphite felt with neutral red (NR) and a modified porous carbon plate with cellulose acetate and porous ceramic membrane, respectively. The air anode operates as a catalyst to generate protons and electrons from water. The growth and ethanol production of Z. mobilis were 50% higher in the ECB than were observed under anoxic nitrogen conditions. Ethanol production by growing cells and the crude enzyme of Z. mobilis were significantly lower under aerobic conditions than under other conditions. The growing cells and crude enzyme of Z. mobilis did not catalyze ethanol production from pyruvate and acetaldehyde. The membrane fraction of crude enzyme catalyzed ethanol production from glucose, but the soluble fraction did not. NADH was oxidized to $NAD^+$in association with $H_2O_2$reduction, via the catalysis of crude enzyme. Our results suggested that NADH/$NAD^+$balance may be a critical factor for ethanol production from glucose in the metabolism of Z. mobilis, and that the metabolic activity of both growing cells and crude enzyme for ethanol fermentation may be induced in the presence of glucose.

• Korean Journal of Microbiology
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• v.23 no.1
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• pp.56-63
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• 1985

We have characterized the plasmids of zymomonas, and constructed E. coli-Zymomonas shuttle vector. Plasmids have been detected in four strains of Zymomonas mobilis. All strains tested had at least one plasmid ranging in size from about 1.7 to 46kb. Antibiotics resistances of Z. mobilis were tested to select the host strain. All strains were very sensitive to tetracycline and chloramphenicol. Homology tests between the plasmids in four strains showed that the plasmids of ATCC10988 is highly homologous to those of ZM1, and that there is no homology between plasmids of ZM4 and Agll. The 1.7kb plasmid of ATCC10988, named as pZM886, also has no homology with plasmids of ZM4. A hybrid plasmid, designated to pBZ41, was constructed from pZM886 and pBR322. A restriction map of pBZ41 was established. Replicon of pZM886 didn't operate in E.coli and pBR322 seemed not to replicate in Zymomonas. pBZ41 was transfered from E. coli to Zymomonas by conjugal mobilization. The transconjugants were resistant to tetracycline and maintained pBZ41 stably.

• Korean Chemical Engineering Research
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• v.46 no.5
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• pp.977-982
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• 2008

The production of ethanol by microbial fermentation as an alternative energy source has been of interest because of increasing oil price. Saccharomyces cerevisiae and Zymomonas mobilis are two of the most widely used ethanol producers. In this study, characteristics of ethanol fermentation by Saccharomyces cerevisiae CHY1077 and Zymomonas mobilis CHZ2501 was compared. Brown rice, naked barley, and cassava were selected as representatives of the starch-based raw materials commercially available for ethanol production. The volumetric ethanol productivities by Saccharomyces cerevisiae from brown rice, naked barley and cassava were $0.68g/l{\cdot}h$, $1.03g/l{\cdot}h$ and $1.28g/l{\cdot}h$ respectively. But for the Zymomonas mobilis, $2.19g/l{\cdot}h$(brown rice), $2.60g/l{\cdot}h$(naked barley) and $3.12g/l{\cdot}h$(cassava) were obtained. Zymomonas mobilis was more efficient strain for ethanol production than S. cerevisiae.

• Genomics & Informatics
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• v.1 no.1
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• pp.55-60
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• 2003

The nucleotide sequence of pZMO1, a small cryptic plasmid of Zymomonas mobilis ATCC10988 was determined. Analysis of 1,680 bp of sequence revealed $69\%$ identity with Shigella sonnei plasmid, pKYM and $61\%$ identity with Nostoc sp. ss DNA replicating plasmid. Analysis of a deduced amino acid sequence of an orf of pZMO1 revealed $75\%$ identity and $90\%$ similarity with the repA gene of Synechocystis sp. plasmid pCA2.4. The upstream region of the repA gene of pZMO1 possesses six directed repeat sequences and two inverted repeat sequences at downstream of the IR consensus sequence of nick region of rolling circle replication (RCR) plasmid. A typical terminator hairpin structure was found at the downstream region of repA gene. Degradation of single-stranded plasmid DNA by S1 nuclease was detected by Southern hybridization. It suggests that pZMO1 replicates by a rolling circle mechanism in Z. mobilis ATCC10988 cells.

• Microbiology and Biotechnology Letters
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• v.15 no.5
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• pp.319-327
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• 1987

In order to develop useful plasmid vectors for Zymomonas cells, attempts were made to isolate natural plasmids from Z. mobilis ATCC10988. Among a few plasmids isolated, a small plasmid of 3.9 Kb size was chosen and designated as pZM3. By introducing the replication origin of pZM3 into pBR325, a hybrid plasmid vector of 8.4 Kb size, pHZ22, was constructed. This vector contained chloramphenicol resistant gene as a selectable marker and proved to be conjugally transmissible and stably maintained in Z. mobilis. Tetracycline resistant gene was isolated from RP4 and introduced into pHZ22 to make a new vector called pHZT224 of 10.7 Kb size. Through n series of experiments, it was evident that these plasmid vectors containing selectable markers of chloramphenicol and tetracycline resistance were shuttle vectors functional in Z. mobilis as well as E. coli.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.94-100
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• 2010

Zymomonas mobilis was immobilized in a modified graphite felt cathode with neutral red (NR-cathode) and Saccharomyces cerevisiae was cultivated on a platinum plate anode. An electrochemical redox reaction was induced by 3 volts of electric potential charged to the cathode and anode. The Z. mobilis produced 1.3-1.5 M of ethanol in the cathode compartment, whereas the S. cerevisiae produced 1.7-1.9 M in the anode compartment after 96 h. The ethanol produced by the Z. mobilis immobilized in the NR-cathode and S. cerevisiae cultivated on the platinum plate was 1.5-1.6 times higher than that produced under conventional conditions. The electrochemical oxidation potential inhibited Z. mobilis, but activated S. cerevisiae. The SDS-PAGE pattern of the total soluble proteins extracted from the Z. mobilis cultivated under the electrochemical oxidation conditions was gradually simplified in proportion to the potential intensity. Z. mobilis and S. cerevisiae were cultivated in the cathode and anode compartments, respectively, of an electrochemical redox combination system. The Z. mobilis culture cultivated in the cathode compartment for 24 h was continuously transferred to the S. cerevisiae culture in the anode compartment at a rate of 300 ml/day. Approx. 1.0-1.2 M of ethanol was produced by the Z. mobilis in the cathode compartment within 24 h, and an additional 0.8-0.9 M produced by the S. cerevisiae in the anode compartment within another 24 h. Thus, a total of 2.0-2.1 M of ethanol was produced by the electrochemical redox combination of Z. mobilis and S. cerevisiae within 48 h.

• KSBB Journal
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• v.26 no.3
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• pp.243-247
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• 2011

Using Bacillus subtilis spore display system, with cotG as an anchoring motif, levansucrase from Zymomonas mobilis, was displayed on the outer surface of Bacillus subtilis spore. Flow cytometry of DB104 (pSDJH-cotG-levU) spore, proved the surface localization of CotG-LevU fusion protein on the spore compared to that of DB104. Enzymatic activity of DB104 (pSDJH-cotG-levU) spore showed more than 1.5 times higher levansucrase specific activity compared to that of the host spore, which is a remarkable increase of enzymatic activity considering the existence of sacA (sucrase) and sacB (levansucrase) in the Bacillus subtilis chromosome. The spore integrity, revealed by sporulation frequency test after heat and lysozyme treatment of spore, did not changed at all in spite of the CotG-LevU fusion protein incorporation into the spore coat layer during spore formation process. These data prove again that Bacillus subtilis spore could be considered as good live immobilization vehicle for efficient bioconversion process.