• 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.

• Microbiology and Biotechnology Letters
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• v.37 no.4
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• pp.350-354
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• 2009

A Gram-negative, alginate lyase-producing bacterium was isolated from the Haeundae Coast, Korea. The isolated strain N7151-6 produced alginate lyase. The optimal temperature and pH for growth were found to be $30^{\circ}C$ and pH 8.0, respectively. This strain can be grown at the NaCl concentration of 0-7% (w/v). Analysis of 16S rDNA sequence and physiological profiling indicated that the strain N7151-6 belonged to Pseudomonas sp. The enzyme alginate lyase produced by Pseudomonas sp. N7151-6 was partially purified by ultrafiltration (MWCO= 30 kDa). The optimum pH and temperature for the activity of the purified enzyme were found to be 7.0 and $30^{\circ}C$, respectively. The enzyme was stable at the pH range of 5.0-9.0 and temperature range of $23-30^{\circ}C$. The total activity of alginate lyase produced was reached about 110 unit/L.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.194-197
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• 1986

Malonate kinase and isocitrate lyase were induced in Acinetobacter calcoaceticus grown on malonate as a sole carbon source but repressed by succinate. The induction of those two enzymes was stimulated by dibutyryl cyclic adenosine 3', 5'-monophosphate, indicating that the expression of their genes for those enzymes is dependent on cyclic adenosine 3', 5'-monophosphate.

• Journal of Microbiology and Biotechnology
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• v.23 no.6
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• pp.872-877
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• 2013

In a previous study, we isolated and reported a second species of the Saccharophagus genus, Saccharophagus sp. strain Myt-1. In the present study, an alginate lyase gene (algMytC) from the genomic DNA of Myt-1 was cloned and characterized. The DNA sequence fragment obtained contained an open reading frame of 1,032 bp that encoded a protein of 343 amino acids with an estimated molecular mass of 37.6 kDa and a pI of 6.60. The deduced protein, AlgMytC, had the conserved amino acid sequences (RTELREM, QIH, YFKAGVYNQ) of the polysaccharide lyase family 7. A BLAST homology search indicated that AlgMytC shared an amino acid sequence identity of 95.9% with alg7A of S. degradans 2-40. The cloned and purified AlgMytC protein showed optimal activity at $40^{\circ}C$, and retained more than 90% of its total activity even after treatment at $25^{\circ}C$ for 24 h. AlgMytC was very alkaliphilic with an optimal pH of 9.0, and more than 90% of its activity was retained in the pH range 8.5-10.0. Moreover, AlgMytC was stable over a wide pH range. The activity of AlgMytC was also stable in the presence of various detergents.

• 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.

• Korean Journal of Microbiology
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• v.31 no.3
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• pp.230-236
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• 1993

The isocitrate lyase extracted from Micrococcus luteus was purified 38.8 folds with the overall yield of 10.2%, by the ammonium sulfate fractionation, DEAE-cellulose, 1st Sephadex G-200 and 2nd Sephadex G-200 column chromatography. The purified enzyme showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified enzyme was estimated 60,000 by the SDS-polyacry]amide gel electrophoresis. The apparent Michaelis constant, Km value for isocitrate was 0.95 mM. The optimum pH and temperature of the purified enzyme were pH 7.5 and $40^{\circ}C$, respectively. The enzyme was activated by $Mg^{2+}$ and inhibited by $Mn^{2+}$, $Ca^{2+}$, $Cu^{2+}$, $Zn^{2+}$ and $CO^{2+}$. In addition, the activity of isocitrate lyase was increased by glutathione and 2-mercaptocthanol at 5 mM and cysteine at I mM.

• Applied Chemistry for Engineering
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• v.23 no.1
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• pp.100-104
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• 2012

Oligosaccharides production showed various biological activities in vivo like functional foods and industrial materials utilized available within many practical applications which have obtained from the degradation of alginate. Alginate is rich in the main component of seaweeds especially the brown algae. We investigated what degrading alginate from seaweeds to make alginate oligosaccharides can utilize in various fields using enzyme secreting Erwinia tasmaniensis. In this study, we observed an optimal culture condition of E. tasmaniensis, and characteristics of alginate lyase secreting E. tasmaniensis. These bacteria, E. tasmaniensis, were isolated from Nuruk. In this case, a suitable growth factor for E. tasmaniensis was culture it for 36 h in broth media on concentration of 1.0% (w/v) alginate. The enzyme showed the highest level of alginate lyase activity when cultured on broth media containing 1.0% (w/v) sodium alginate for 72 h. Optimal condition of pH, temperature and duration time for alginate lyase activity were found to be pH 6.0, $20^{\circ}C$ and 60 min, respectively.

• Microbiology and Biotechnology Letters
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• v.15 no.2
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• pp.80-83
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• 1987

During the enzymatic production of L-phenylalanine exploiting L-phenylalanine ammonia lyase(E.C 4.3.1.5) and trans-cinnamic acid, the conversion yield of L-phenylalanine and the stability of L-phenylalanine ammonia-lyase per so or induced Rhodotorula glutinis IFO 0559 were investigated. And the glycerol added to the conversion reaction as stabilizer had effect only on L-phenylalanine and made it possible to obtain the 80％ conversion yield from trans-cinnamic acid. In addition, the more rapid and reliable method than the thin layer chromatography for determining the conversion yield will be disscused.

• Development and Reproduction
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• v.10 no.2
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• pp.127-133
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• 2006

[ $17\;{\alpha}-hydroxylase/17,20-lyase(P450_{C17})$ ] is the key enzyme mediating the conversion of progesterone to $17\;{\alpha}-hydroxyprogesterone$, ultimately to androstenedione during steroidogenesis. R. dybowskii's ovarian $P450_{C17}$ cDNA was cloned to understand the regulatory mechanism of ovarian steroidogenic pathway at the molecular level in amphibian. A 2.5kb cDNA clone encoding a single open-reading frame with a 519 deduced amino acid was isolated with the screening of ovarian cDNA library. This sequence contained the three highly conserved domains as seen in $P450_{C17}$ of other species. The comparison of amino acid sequence of Rana $P450_{C17}$ with other animal's $P450_{C17}$ showed relatively high identity with 76% in Xenopus, 63% in chicken, 60% in rainbow trout, and 45% in human. Phylogenic analysis also indicated that Rana $P450_{C17}$ gene was evolutionary well conserved among vertebrate. Northern analysis indicated that the two different sizes of $P450_{C17}$ transcripts with approximately 2.5 and 3.6kb were detected in ovary tissue, but not in other tissues. The expression vector of Rana $P450_{C17}$ clearly showed the $17\;{\alpha}-hydroxylase$ activity converting the exogenous progesterone into $17\;{\alpha}-hydroxyprogesterone$ in the nonsteroidogenic COS-1 cells. Therefore, Rana $P450_{C17}$ cDNA is very useful to investigate the molecular mechanism of the ovarian steroidogenesis in amphibian.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.625-633
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• 2007

We isolated a new extracellular alginate lyase-producing microorganism, which displayed alginate-depolymerizing activity in plate assays, from coastal soils in Wando, Jeollanam-do, Korea. This alginate-depolymerizing bacterium belonged to the genus Streptomyces and it was named Streptomyces sp. MET 0515. An extracellular alginate lyase(ALY1) secreted by Streptomyces sp. MET 0515, was purified to homogeneity by a combination of acetone precipitation, anion-exchange chromatography (Q-Sepharose and DEAE-Sepharose) and Sephacryl S-200 HR gel filtration chromatography. Its molecular mass was 26 kDa as determined by SDS-PACE analysis. The enzyme had an optimal temperature of $70^{\circ}C$ for its activity, and was most active at pH 7.5. The thermal and pH stability were $0-50^{\circ}C$, and pH 6.0-9.0, respectively. The enzyme activity was stimulated by 1mM $Mn^{2+}$, and inhibited by 1mM $Fe^{3+}$, 1mM EDTA and 1mM $Zn^{2+}$. Preliminary analysis of substrate specificity showed that this alginate lyase had activity on both poly-alpha 1,4-L-guluronate and poly-beta 1,4-D-mannuronate in the alginate molecule.