• Journal of Environmental Science International
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• v.7 no.6
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• pp.853-857
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• 1998

Purification of the isocitrate lyase extracted from Microbacterium laevaniformans was investigated. The isocitrate lyase was purified 43.6 folds by the following continuous treatment with ammonium sulfate fraction, DEAE-cellulose, DEAE-sephacel and Sephadex G-200 chromatography. The purified isocitrate lyase was showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified isocitrate lyase was estimated 54,000 Da by the SDS-polyacrylamide gel electrophoresis. The Km and Vmax values for isocitrate were estimated to be 0.83mM and 0.33units/ml, respectively. Activity of isocitrate lyase was inhibited by cystein-HCl and glutathione.

• Journal of Life Science
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• v.19 no.11
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• pp.1522-1528
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• 2009

A marine bacterium was isolated from brown seaweeds for its ability to degrade alginate. Analysis of 16S ribosomal DNA sequence revealed that the strain belongs to Streptomyces like strain ALG-5 which was reported previously. New alginate lyase gene of Streptomyces sp. M3 was cloned by using PCR with the specific primers designed from homologous nucleotide sequences. The consensus sequences of N-terminal YXRSELREM and C-terminal YFKAGXYXQ were conserved in the M3 alginate lyase amino acid sequences. The homology model for the M3 alginate lyase showed a characteristic structure of $\beta$-jelly roll fold main domain like alyPG from Corynebacterium sp. ALY-1. The homogenate of the recombinant E. coli with the alginate lyase gene showed more degrading activity for polyguluronate block than polymannuronate block. The results from the multiple alignments and the homology modeling elucidated in the M3 alginate lyase can be classified into family PL-7.

• Korean Journal Plant Pathology
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• v.10 no.3
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• pp.157-162
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• 1994

Erwinia rhapontici causes soft-rot disease in a number of plants such as onion, garlic and hyacinth. There has been no report that E. rhapontici produces pectate lyase. Pel gene was cloned from genomic DNA of the parasitic soft-rot E. rhapontici polymerase chain reaction by using synthetic oligonulceotide primers designed from the pel 1 to E. carotovora. The recombinant plasmid pJE101 containing pectate lyase gene, when introduced into E. coli DH5$\alpha$, produced pectate lyase an macerated hyacinth tissue.

• Journal of Life Science
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• v.20 no.11
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• pp.1582-1588
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• 2010

A new alginate lyase gene of marine bacterium Streptomyces sp. M3 had been previously cloned in pColdI vector and transformed into E. coli BL21 (DE3). In this study, M3 lyase protein without signal peptide was overexpressed by induction with IPTG and purified with Ni-Sepharose affinity chromatography. The absorbance at 235 nm of the reaction mixture and TLC analysis showed that M3 alginate lyase was a polyG-specific lyase. When M3 lyase was assayed with substrate for 10 min, optimum pH and optimum temperature were pH 9 and $60^{\circ}C$. For the effect of 1mM metal ion on M3 lyase activity, $Ca^{++}$ and $Mn^{++}$ ions increased the alginate degrading activity by two-fold, whereas $Hg^{++}$ and $Zn^{++}$ ions inhibited the lyase activity completely. $Mg^{++}$, $Co^{++}$, $Na^+$, $K^+$, and $Ba^{++}$ did not show any strong effects on alginate lyase activity.

• Korean Journal of Food Science and Technology
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• v.29 no.2
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• pp.314-319
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• 1997

The objectives of this study were to examine the effect of storage time, temperature, pH, and NaCl concentration on hydroperoxide lyase and lipoxygenase activities, and to establish important informations to the production of typical cucumber flavor. Conditions affecting lipoxygenase and hydroperoxide lyase would be important for cucumber flavor production. Maximum activity was observed at pH 5.0 for hydroperoxide lyase and pH 5.5 for lipoxygenase. Both enzymes were relatively stable at $40\;to\;50^{\circ}C$ for 6days storage time. Maximum activity of both enzymes was observed with 0.2 M NaCl at pH 5.0. Activities were stimulated with concentrations of NaCl from 0.05 to 0.2 M. Hydroperoxide lyase and lipoxygenase activities were decreased at concentration of NaCl greater than 0.2 M.

• Microbiology and Biotechnology Letters
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• v.22 no.1
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• pp.65-72
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• 1994

Purification and characterization of pectate lyase from Bacillus sp HSA-925. Bacillus sp. HSA-925 isolated from soil produced constitutively an extracellular pectate lyase when cultivated in LB broth. The pectate lyase(EC 4.2.2.2) was purified from the cuylture broth by preciptation with ammonium sulfate, followed by column chromatography on CM-cellulose C-50 and repeated gel filtration on Sephadex G-75G. The enzyme had a molecular weight of 32-33 kDa. The activity was mazimum at pH 9.5 AND 45$\CIRC $C. The enzume activity was stable at 55$\circ $C for 15 min and between pH7-12. The activation energy, Km and V$_{max}$ for the pectate lyase were 5.8779 kcal/mol, 6.33$\times $10$^{-2}$ mol/ml and 2.09$\times $10$^{2}$ $\mu $mol/min respectively. The enzyme was activated by Ca$^{2+}$, Cu$^{2+}$ and inhibited by Li$^{+}$, Hg$^{2+}$, EDTA.

• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.420-424
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• 1987

Isocitrate lyase from crude extract of Saccharomycopsis lipolytica ATCC44601 and MX9-11RX8 temperature-sensitive mutant was purified about 54 times and 87 times, respectively by ammonium sulfate fractionation, Toyo peal HW-55F gel filtration and DEAE-Cellulose ion exchange chromatography, The molecular weight of the purified isocitrate lyase from this yeast was estimated to be 230, 000 by gel filtration on Sephadex G-200, and SDS-polyacrylamide Eel electrophoresis showed that the enzyme consisted of four identical or similar subunits with a molecular weight of 59, 000 and the enzyme showed optimum activity at pH 6.9.

• Korean Journal Plant Pathology
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• v.12 no.1
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• pp.129-131
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• 1996

Pecatate lyase(Pel)는 펙틴과 펙틴산을 분해하며, 기주식물의 감염에 관여한다. Erwinia rhapontici에 있어서 기주와 병원균이 병원성과의 상호관계를 구명하기 위하여 pectate lyase(Pel) 활성에 미치는 식물체 추출물과 세포벽의 효과를 검토하였다. 본 균은 glycerol이 포함된 minimal salts(MSG) 배지와 식물체 추출물이 첨가된 MSP 배지에서는 Pel 활성이 검출되지 않았다. 그러나 배추, 상추 잎, 감자 괴경, 셀러리 잎자루, 양파 인경, 당근 뿌리의 세포벽이 첨가된 MSP 배지에서는 Pel의 활성이 검출되었다. Pel을 유도하는 식물 인자는 불용성이고, 열처리에 불안전하였다.

• Journal of Life Science
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• v.24 no.2
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• pp.128-136
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• 2014

Alginates are linear acidic polysaccharides composed with (1-4)-linked ${\alpha}$-L-guluronic acid and ${\beta}$-Dmannuronic acid. Alginate can be degraded by diverse alginate lyases, which cleave the alginate using a ${\beta}$-elimination reaction and produce unsaturated uronate oligomers. A gene for a polyMG-specific alginate lyase possessing a novel structure was previously identified and cloned from Stenotrophomonas maltophilia KJ-2. Homology modeling of KJ-2 polyMG-specific alginate lyase showed it belongs to the PL6 family, whereas three Azotobacter vinelandii polyMG lyases belong to the PL7 family of polysaccharide lyases. From $^1H$-NMR spectra data, KJ-2 polyMG lyase preferably degraded the M-${\beta}$(1-4)-G glycosidic bond than the G-${\alpha}$(1-4)-M glycosidic bond. Seventeen mutants were made by site-directed mutagenesis, and alginate lyase activity was analyzed. Lys220Ala, Arg241Ala, Arg241Lys, and Arg265Ala lost alginate lyase activity completely. Arg155Ala, Gly303Glu, and Tyr304Phe also lost the activity by 60.7-80.1%. These results show that Arg155, Lys220, Arg241, Arg265, Gly303, and Tyr304 are important residues for catalytic activity and substrate binding.

• KSBB Journal
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• v.26 no.6
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• pp.538-542
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• 2011

The research was purposed production of oligosaccharide from alginate hydrolysis the main composition in cell walls of sea weed. We was isolated 252 strains from sea water and mud flat, the highest alginate lyase activity was selected, and identified as Sanguibacter keddieii NC9 by 16S rDNA sequence analysis. In this study was select the sodium alginate concentration, pH, temperature for the production of alginate lyase activity. Alginate lyase activity was confirmed from plate assay with 10% cetylpyridinium chloride. The optimum culture conditions for the production of alginate lyase were sodium alginate 10 g/L, peptone 5 g/L, $40^{\circ}C$, pH 9 and 36 hours incubation time. Sanguibacter keddieii NC9, its alginate lyase would be useful for the production of bioenergy and biofunctional oligosaccharides from sea weed.