• Journal of Applied Biological Chemistry
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• v.53 no.2
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• pp.105-107
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• 2010

To overcome the limitation of E. coli expression system such as inclusion body formation and disulfide bond failure, we tried to express the heterologous protein as a secreted form. We adopted agarase signal peptide (ASP; 23 amino acid residues) from Agarivorans albus YKW-34 which is one of marine bacteia. When we used ASP to express $\beta$-agarase, about 42% activity was detected in media.

• KSBB Journal
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• v.21 no.5
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• pp.389-393
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• 2006

Characteristics of ${\beta}$-agarase production of Agarivorans sp, JA-1 isolated from north-eastern sea of Jeju marine environment was studied. Optimal cell growth was definite that the medium containing agar is 0.2%. The decreasing pattern of viscosity and agar concentration was same and they reached almost zero after 15 hours. Fed-batch culture was studied to improve agarase productivity by Agarivorans sp. JA-1 in marine broth containing 2.0 g/L agar with intermittent addition of 0.8 g agar two times. The hydrolysis products were identified oligosaccharide of degrees of polymerization 6.

• Journal of Microbiology and Biotechnology
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• v.22 no.9
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• pp.1237-1244
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• 2012

Agarase genes of non-marine agarolytic bacterium Cellvibrio sp. were cloned into Escherichia coli and one of the genes obtained using HindIII was sequenced. From nucleotide and putative amino acid sequences (713 aa, molecular mass; 78,771 Da) of the gene, designated as agarase AgaA, the gene was found to have closest homology to the Saccharophagus degradans (formerly, Microbulbifer degradans) 2-40 aga86 gene, belonging to glycoside hydrolase family 86 (GH86). The putative protein appears to be a non-secreted protein because of the absence of a signal sequence. The recombinant protein was purified with anion exchange and gel filtration columns after ammonium sulfate precipitation and the molecular mass (79 kDa) determined by SDS-PAGE and subsequent enzymography agreed with the estimated value, suggesting that the enzyme is monomeric. The optimal pH and temperature for enzymatic hydrolysis of agarose were 6.5 and $42.5^{\circ}C$, and the enzyme was stable under $40^{\circ}C$. LC-MS and NMR analyses revealed production of a neoagarobiose and a neoagarotetraose with a small amount of a neoagarohexaose during hydrolysis of agarose, indicating that the enzyme is a ${\beta}$-agarase.

• Fisheries and Aquatic Sciences
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• v.13 no.1
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• pp.36-48
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• 2010

An agar-degrading Agarivorans sp. AG17 strain was isolated from the red seaweed Grateloupia filicina collected from Jeju Island. A beta-agarase gene from Agarivorans sp. AG17 was cloned and designated as agrA. agrA has a 2,985 bp coding region encoding 995 amino acids and was classified into the glycoside hydrolase family (GHF)-50. Predicted molecular mass of the mature protein was 105 kDa. His-tagged agrA was overexpressed in Escherichia coli and purified as a fusion protein. The enzyme showed 158.8 unit/mg specific activity (optimum temperature at $65^{\circ}C$ and pH 5.5 in acetate buffer) with unique biochemical properties (high thermal and pH stabilities). Enzyme produced neoagarohexaose, neoagarotetraose and neoagarobiose by degrading agar, and hydrolyzed neoagaro-oligosaccharides were biologically active. Hence the purified enzyme has potential for use in industrial applications such as the development of cosmetics and pharmaceuticals.

• Proceedings of the Korean Society of Life Science Conference
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• 2008.10a
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• pp.55.1-55.1
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• 2008

• Microbiology and Biotechnology Letters
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• v.52 no.2
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• pp.135-140
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• 2024

This study reports the isolation of a bacterium capable of degrading agar and the characterization of its agarase. An agar-degrading marine bacterium JS-1 was isolated using Marine agar 2216 media from seawater collected from the seashore of Angolpo, Changwon, Gyeongnam Province, Republic of Korea. An agar-degrading bacterium was named as Tenacibaculum sp. JS-1 by phylogenetic analysis based on 16S rRNA gene sequence. The extracellular crude agarase was prepared from the culture media of Tenacibaculum sp. JS-1 and used for characterization. Relative activities at 20, 30, 40, 50, and 60℃ were 39, 73, 100, 74, and 53%, respectively. Relative activities at pH 5, 6, 7, and 8 were 46%, 67%, 100%, and 49%, respectively. Its extracellular agarase showed maximum activity (164 U/l) at pH 7.0 and 40℃ in a 20 mM GTA buffer. The residual activities after heat treatment at 20, 30, and 50℃ for 30 min were 84, 73, and 26% or more, respectively. After 2 h heat treatment at 20, 30, 40, and 50℃, the residual activities were 80, 64, 52 and 21%, respectively. Thin layer chromatography analysis suggested that Tenacibaculum sp. JS-1 produces extracellular β-agarases that hydrolyze agarose to produce neoagarooligosaccharides, including neoagarohexaose (12.3%), neoagarotetraose (65.1%), and neoagarobiose (22.6%) at 6 h. Tenacibaculum sp. JS-1 and its β-agarase could be valuable for producing neoagarooligosaccharides with a variety of functional properties. These properties include inhibiting bacterial growth, slowing down starch degradation, and whitening, which are of interest for pharmaceuticals, food, cosmeceuticals, and nutraceuticals.

• Proceedings of the Korean Society of Life Science Conference
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• 2005.09a
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• pp.72-72
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• 2005

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.776-783
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• 2018

The agarase gene gaa16a was identified from a draft genome sequence of Gilvimarinus agarilyticus JEA5, an agar-utilizing marine bacterium. Recently, three agarase-producing bacteria, G. chinensis, G. polysaccharolyticus, and G. agarilyticus, in the genus Gilvimarinus were reported. However, there have been no reports of the molecular characteristics and biochemical properties of these agarases. In this study, we analyzed the molecular characteristics and biochemical properties of agarases in Gilvimarinus. Gaa16A comprised a 1,323-bp open reading frame encoding 441 amino acids. The predicted molecular mass and isoelectric point were 49 kDa and 4.9, respectively. The amino acid sequence of Gaa16A showed features typical of glycosyl hydrolase family 16 (GH16) ${\beta}$-agarases, including a GH16 domain, carbohydrate-binding region (RICIN domain), and signal peptide. Recombinant Gaa16A (excluding the signal peptide and carbohydrate-binding region, rGaa16A) was expressed as a fused protein with maltose-binding protein at its N-terminus in Escherichia coli. rGaa16A had maximum activity at $55^{\circ}C$ and pH 7.0 and 103 U/mg of specific activity in the presence of 2.5 mM $CaCl_2$. The enzyme hydrolyzed agarose to yield neoagarotetraose as the main product. This enzyme may be useful for industrial production of functional neoagaro-oligosaccharides.

• Journal of Food Hygiene and Safety
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• v.15 no.4
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• pp.277-281
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• 2000

Agar, one of the most abundant marine products has not been utilized extensively because of low level of processing technology in Korea. This research was carried out to improve the utilization of agar and consequent increase in profit. Antibacterial activity of agarooligosaccharides were evaluated against bacteria causing putrefaction and flood poisoning. Addition of 0.4% agarooligosaccharides showed antibacterial activity toward Staphylococcus aureus and Escherichia coli O157:H7; furthermore, autoclave treatment of agarooligosaccharides solution enhanced the antibacterial activity. Agarooligosaccharides showed high stability against the pH change. Addition of amino acid(alanine, lysine, glycine, phenylalanine) in agarooligosaccharides solution enhanced antibacterial activity in E. coli O157:H7, Streptococcus mutans and Staphylococcus aureus.

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1622-1628
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• 2014

DagA, a ${\beta}$-agarase, was produced by cultivating a recombinant Streptomyces lividans in a glucose medium or a mixed-sugar medium simulating microalgae hydrolysate. The optimum composition of the glucose medium was identified as 25 g/l glucose, 10 g/l yeast extract, and $5g/l\;MgCl_2{\cdot}6H_2O$. With this, a DagA activity of 7.26 U/ml could be obtained. When a mixed-sugar medium containing 25 g/l of sugars was used, a DagA activity of 4.81 U/ml was obtained with very low substrate utilization efficiency owing to the catabolic repression of glucose against the other sugars. When glucose and galactose were removed from the medium, an unexpectedly high DagA activity of about 8.7 U/ml was obtained, even though a smaller amount of sugars was used. It is recommended for better substrate utilization and process economics that glucose and galactose be eliminated from the medium, by being consumed by some other useful applications, before the production of DagA.