• KSBB Journal
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• v.11 no.1
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• pp.115-123
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• 1996

Two kinds of mutants were isolated to clone a cluster of genes essential for zooglan biosynthesis. Zoogloea ramigera 115 strains produce capsular polysaccharide. To achieve conjugation in strain 115 and to facilitate recovery of product, a capsule non-forming strain was isolated via successive centrifugation and screening. The other kind of mutants devoid of or producing altered exopolysaccharides were obtained using classical transposon(Tn5) technique and screened for altered colony morphology and celluflour binding properties. Complementation of these mutants was achieved with Z. ramigera 115 slime gene library constructed in a broad host range cosmid vector and helper plasmid by triparental conjugation.

• Microbiology and Biotechnology Letters
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• v.26 no.1
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• pp.83-88
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• 1998

Heavy metal removal by Z. ramigera 115 and soluble slime polymer producing mutant Z. ramigera 115SLR was investigated. Both strains showed similar tolerance against $Cd^{2+}$, $Co^{2+}$, $Cu^{2+}$, $Ni^{2+}$ and $Fe^{2+}$. When cells were cultivated in the presence of 500 ppm $Cd^{2+}$, the mutant strain removed 1.5 fold more metal than the wild type did at same biomass. Metal adsorption capacities were in the order of Z. ramigera l15SLR polymer > Z. ramigera 115 polymer > Z. ramigera 115 cell >Z. ramigera l15SLR cell. The optimum pH for metal adsorption was 7.5. Langmuir and Freundlich isotherms indicated that Qmax and 1/n of Z. ramigera l15SLR polymer were 164.2 mg $Cd^{2+}$/g dw and 0.496, respectively. These results showed that the polymer of Z. ramigera l15SLR could be used as an effective metal adsorbate.

• Journal of Korean Society of Water and Wastewater
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• v.20 no.1
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• pp.63-70
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• 2006

Biosorption characteristics were investigated at various temperature and pH conditions in order to establish lead(II) removal using Zoogloea ramigera 115SLR. Biosorption equilibrium isotherms and kinetics were obtained from batch experiments. The Freundlich and Langmuir model could be described the biosorption equilibrium of lead(II) on Z. ramigera 115SLR, Ca-alginate bead and immobilized Z. ramigera 115SLR. The maximum biosorption capacity of Z. ramigera 115SLR increased from 325 to 617mg $pb^{2+}/g$ biomass as temperature increased from 288.15 K to 308.15K from the Langmuir model. Fixed-bed column breakthrough curves for lead(II) removal were also obtained. For regeneration of the biosorbent, complete lead(II) desorption was achieved using 5mM HCl in fixed-bed column. This study shows the possibilities that well-treated immobilized Z. ramigera 115SLR with the mechanical intensity like TEOS (Tetraethyl orthosilicate) treatment and the optimum acid solution for desorption can be used for the effective treatment for lead(II) containing wastewater.

• Biomedical Science Letters
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• v.6 no.1
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• pp.1-9
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• 2000

To identity the genes responsible for the biosynthesis of exopolysaccharide, recombinant plasmids pUEX10 and pLEX10 were constructed from plasmid pLEX3 which was isolated from the recombinant cosmid library of Zoogloea ramigera 115. The complete nucleotide sequence of the 1.7 kb genomic DNA insert in plasmid pUEX10 was determined. Its analysis identified two open reading frames (ORF3 & ORF4) which could encode two proteins. The amino acid sequence derived from ORF3 showed the homology with gumC protein in Xanthomonas campestris as well as exoP protein in Rhizobium melizoti. The partial amino acid sequence of ORF4 showed the homology with polysaccharide export protein in Thermotoga maritima. Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 showed the similar pattern for EPS production. Yield of exopolysaccharides produced by Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 was 0.26% (w/v) and 0.16% (w/v), respectively.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.161-168
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• 2000

Plasmid pLEX3 isolated from the recombinant cosmid library of Zoogloea ramigera 115 was found to be responsible for the restoration of the rugose colony phenotype. To confirm the essential region responsible for the complementation, subclones were constructed from plasmid pLEX3 and transformed into mutant strain Z. ramigera 115SLR. The recombinant plasmids pLEX10 and pLEX11 were shown to complement the slime-forming property of Z. ramigera 115SLR. In a compositional analysis of the exopolysaccharides from Z. ramigera 115, Z. ramigera 115SLR, and Z. ramigera 115SLR harboring plasmid pLEX11, the exopolysaccharides showed a similar composition with glucose, galactose, and side chain groups. The complete nucleotide sequence of the 3.25kb genocim DNA insert in plasmid pLEX11 was determined and its analysis identified two open reading frames which could encode two proteins. The gene products derived form the two open reading frames were confirmed by and in vivo transcription using a T7-RNA polymerase. The ORF1 produced a 30 kDa protein, whereas the ORF2 was found responsible for the complementation of the morphological mutation and produced a 14 kDa protein. An in vivo gene expression of plasmid pTEX10 showed another open reading frame encoding a 50 kDa protein. The gene products form ORF1 and ORF2 are regarded as novel proteins which do not show any homology with other proteins.

• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.1163-1168
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• 2006

The recombinant plasmid pLEX2FP complements the mutation in Zoogloea ramigera 115MM1, and the complemented mutant produces an exopolysaccharide that shows higher affinity for the calcofluor dye than the exopolysaccharide from Z. ramigera 115SLR, resulting in higher fluorescence intensity under UV light. A compositional and structural analysis of the exopolysaccharide from Z. ramigera 115MM1 showed that the different fluorescent properties were due to a lower content of acetyl groups when compared with Z. ramigera 115SLR exopolysaccharide. These results were in agreement with a sequence analysis of the gene carried in the plasmid pLEX2FP, which appeared to encode an O-acyltransferase highly homologous to the 3-O-acyltransferase of Streptomyces mycarofaciens. The gene encoding the acyltransferase from Z. ramigera 115SLR was expressed as a GST-fusion protein with 70,000 daltons in E. coli.

• Journal of Microbiology and Biotechnology
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• v.6 no.5
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• pp.321-325
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• 1996

Mutants having altered levels and/or types of EPS in exopolysaccharide biosynthesis were isolated after NTG mutagenesis of Zoogloea ramigera 115SLR. Mutant candidates were classfied with five groups based on the observed characteristics for EPS biosynthesis pattern. The recombinant plasmids pLEX3BS and pLEX3BM were constructed from pEX3B which was previously isolated from genomic DNA of Z. ramigera 115SLR. Plasmid pLEX3BM with a 7.8 kb insert DNA contains an additional 1.8kb DNA fragment which is not present in pLEX3BS containing 13 kb insert DNA. Plasmid pLEX3BM was able to complement the mutation responsible for the changes in morphology of Z. ramigera 115SLR. However, the complementation of EPS negative mutant strains was not successful with pLEX3BM. Plasmid pLEX3BS on the other hand complemented the mutation responsible for the loss of EPS biosynthesis, resulting in the restoration of Z. ramigera 115SLR phenotype. But this plasmid was not able to complement the morphological mutant strain, Z. ramigera 115SLR.

• KSBB Journal
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• v.7 no.3
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• pp.166-171
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• 1992

Zoogloea ramigera 115 was cultured for biopolymer production and its bioflocculant usages. Cultural conditions of the organism were examined with regard to high production of the microbial polysaccharide. The most suitable medium was found to contain glucose as a carbon source, $NaNO_3$ as a nitrogen source, and yeast extract as an organic nutrient. The initial pH of 6.0 proved to optimal. The biopolymer was extracted effectively using ultrasonication and high speed centrifugation, followed by propanol addition. Jar test results indicate that the polysaccharide produced by the organism is an effective flocculant.

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.196-202
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• 1992

Zoogloea ramigera 115 was selected for the production of viscous microbial polysaccharide for bioflocculants usage. Batch, fed-batch, and continuous culture processes were examined with regard to the high biopolymer production. Several carbon sources were tested, including glucose, lactose, molasses, and cheese whey. The C/N ratio of 90 was most effective for biopolymer production from glucose, while the C/N ratios of 30 for lactose and 60 for both molasses and cheese whey substrate gave a maximum production. Fed batch culture proved more effective to increase final biopolymer concentration than batch culture. Continuous fermentation with two stages modifying C/N ratio increased the productivity. The production rates were a maximum at dilution rate of 0.048 $hr^{-1}$ for molasses and at 0.096 $hr^{-1}$for cheese whey.

• Microbiology and Biotechnology Letters
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• v.30 no.3
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• pp.264-269
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• 2002

The production of exopolysaccharide from Zoogloea ramigera 115SLR in batch culture was affected by carbon sources, rifampicin concentration, inoculum size and cell density of starter culture. The increase of organic nitrogen concentration and cell density in defined medium by adding starter culture resulted in higher production of exopolysaccharide (EPS) within 2 days. Glucose and lactose as carbon sources showed EPS production of 10.7 gk and 10.5 ga, respectively. Higher EPS production was obtained with 2.5% (w/v) glucose. White sugar, brown sugar and galactose produced less than 10 gt of EPS. The optical density of starter culture affected on EPS yield, producing over 10 gt of EPS in the range of 1.0-1.7. In the presence of rifsmpicin, Z. ramigera 115SLR produced EPS of 12.9 gt. Molecular weight of EPS produced with/without rifampicin was determined with 1.367$\times$106 and 1.711$\times$106 g/mol using HPSEC-MALLS-RI, respectively.