• Title/Summary/Keyword: GH-118

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Cloning, Expression, and Characterization of a Glycoside Hydrolase Family 118 ${\beta}$-Agarase from Agarivorans sp. JA-1

  • Lee, Dong-Geun;Jeon, Myong Je;Lee, Sang-Hyeon
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1692-1697
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    • 2012
  • We report a glycoside hydrolase (GH)-118 ${\beta}$-agarase from a strain of Agarivorans, in which we previously reported recombinant expression and characterization of the GH-50 ${\beta}$-agarase. The GH comprised an open reading frame of 1,437 base pairs, which encoded a protein of 52,580 daltons consisting of 478 amino acid residues. Assessment of the entire sequence showed that the enzyme had 97% nucleotide and 99% amino acid sequence similarities to those of GH-118 ${\beta}$-agarase from Pseudoalteromonas sp. CY24, which belongs to a different order within the same class. The gene corresponding to a mature protein of 440 amino acids was inserted, recombinantly expressed in Escherichia coli, and purified to homogeneity with affinity chromatography. It had maximal activity at $35^{\circ}C$ and pH 7.0 and had 208.1 units/mg in the presence of 300 mM NaCl and 1 mM $CaCl_2$. More than 80% activity was maintained after 2 h exposure to $35^{\circ}C$; however, < 40% activity remained at $45^{\circ}C$. The enzyme hydrolyzed agarose to yield neoagarooctaose as the main product. This enzyme could be useful for industrial production of functional neoagarooligosaccharides.

Production and Application of Recombinant Agarase (재조합 한천 분해효소의 생산과 응용)

  • Kim, Se Won;Hong, Chae-Hwan;Yun, Na Kyong;Shin, Hyun-Jae
    • Journal of Marine Bioscience and Biotechnology
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    • v.8 no.1
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    • pp.1-9
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    • 2016
  • The hydrolysis of biomass to fermentable sugar (saccharification) and to oligosaccharide is an essential process in biotechnology including biorefinery and biofood. Various macroalgae are commercially cultivated in several Asian countries as a useful resource for food and agar production. Agar is a major component of the cell walls of red algae that can be hydrolyzed by agarase. Agarases are classified into ${\alpha}$-agarase (E.C. 3.2.1.158) and ${\beta}$-agarase (E.C. 3.2.1.81) according to the cleavage pattern and grouped in the glycoside hydrolase (GH) family (GH-16, GH-58, GH-86, GH-96, and GH-118) based on the amino acid sequences of the proteins. Agarases have been isolated from various bacteria found in seawater and marine sediments. To increase productivity of the enzyme, a research on recombinant enzymes has been done. The application of recombinant agarase can be possible in the various filed such as energy, food, cosmetics, medical and so on. This paper reviews the source, biochemical characteristics and production system of recombinant agarases for further study.

Cloning, Expression, and Characterization of a Novel GH-16 β-Agarase from Agarivorans sp. JA-1 (Agarivorans sp. JA-1 유래 신규 GH-16 β-agarase의 클로닝, 발현 및 특성)

  • Jeon, Myong Je;Kim, A-Ram;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1545-1551
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    • 2012
  • Authors report the glycoside hydrolase (GH) family 16 ${\beta}$-agarase from the strain of Agarivorans sp. JA-1, which authors previously stated as recombinant expression and characterization of GH-50 and GH-118 ${\beta}$-agarase. It comprised an open reading frame of 1,362 base pairs, which encodes a protein of 49,830 daltons consisting of 453 amino acid residues. Valuation of the total sequence showed that the enzyme has 98% nucleotide and 99% amino acid sequence similarities to those of GH-16 ${\beta}$-agarase from Pseudoalteromonas sp. CY24. The gene corresponding to a mature protein of 429 amino acids was recombinantly expressed in Escherichia coli, and the enzyme was purified to homogeneity by affinity chromatography. It showed maximal activity at $40^{\circ}C$ and pH 5.0, representing 67.6 units/mg. Thin layer chromatography revealed that mainly neoagarohexaose and neoagarotetraose were produced from agarose. The enzyme would be valuable for the industrial production of functional neoagarooligosaccharides.

The Classification, Origin, Collection, Determination of Activity, Purification, Production, and Application of Agarases (Agarase의 분류, 기원, 확보, 활성파악, 분리정제, 생산 및 응용)

  • Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.22 no.2
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    • pp.266-280
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    • 2012
  • Agar is a cell wall component of macro red algae that can be hydrolyzed by agarase. Agarases are classified into ${\alpha}$-agarase (E.C. 3.2.1.158) and ${\beta}$-agarase (E.C. 3.2.1.81), in accordance with their cleavage pattern, and can be grouped in the glycoside hydrolase (GH)-16, -58, -86, -96, and -118 family according to the amino acid sequences of the proteins. Many agarases and/or their genes have been detected, isolated, and recombinantly expressed from bacteria, and metagenomes have their origins in sea and terrestrial environments. Products of agarases, agarooligosaccharides and neoagarooligosaccharides, represent wide functions such as antitumor, immune stimulation, antioxidation, prebiotic, hepa-protective, antibacterial, whitening, and moisturizing effects; hence, broad applications would be possible in the food industry, cosmetics, and medical fields. In addition, agarases are also used as a tool enzyme for research. This paper reviews the sources, purifications and detection methods, and application fields of agarases. The role of agarases in agar metabolism and the function of their enzymatic products are also surveyed.

Analysis on the Occurrence Factors of High-Risk Diseases of Pregnant Women by the Degree of Obesity (산모의 비만정도에 따른 고위험 질환 발생요인 분석)

  • Kim, Su-Min;Ye, Soo-Young
    • Journal of the Institute of Convergence Signal Processing
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    • v.19 no.3
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    • pp.118-124
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    • 2018
  • Obecity in pregnancy causes many problems and increases risk of pregnancy complications at the time of childbirth. But there is a lack of comprehensive analysis of factors that are negatively affected during pregnancy. Therefore, this study is intended to analyze seven factors of high-risk maternal diseases by the degree of obesity using body mass index(BMI). We conducted a cross tabulation analysis and regression analysis to analisized relationship between variables : Gestational Hypertension(GH), Gestational Diabetes Mellitus(GDM), Thyroid Stimulation Hormone(TSH), Age, Blood Urea Nitrogen Test(BUN), Total-Cholesterol(T-C), and newborn's weight. As a result, the more the obesity level of mothers increases, the more the proportion of mothers with GH, GDM, TSH increases. And there was a positive relationship between the BMI of mothers and their age, T-C, and Newborn weight, and a negative relationship to the BUN.

The Development of Functional Food with Plant Extracts for Enhancing Growth Rate (생약추출물을 이용한 키 성장 기능성 식품 개발)

  • Ra, Jeong-Chan;Park, Hyeong-Geun;Choi, Mi-Kyung;Lee, Hang-Young;Kang, Kyung-Sun
    • Journal of Food Hygiene and Safety
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    • v.19 no.3
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    • pp.112-118
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    • 2004
  • It has been reported that various kinds of chinese herbs have an activity of promote growth rate in both animals and human. To investigate the growth promoting effect of the selected plants, fish and pigs were used as experimental animals. In fish, Eleutherococcus senticosus extract and Artemisia capillasis extract were found to be most effective. And these plant extracts were given to pigs. The result showed that plant extracts-fed pigs were significantly increased their body weight gain at 7.06% of average daily gain, as compared to control. The verify this results, secreted growth hormone (GH)levels and insulin-like growth factor-1 (ICF-1) levels into blood were measured. This result indicated that GH and IGF-1 levels in the blood in plant extracts-fed pigs were higher that those of control. To confirm growth promotion effect on human, we manufactured the mixture of these plant extracts, and coated this mixture onto rice, named as $Kiwoomi^{TM}$. When we administered $Kiwoomi^{TM}$ to elementary students, it was found to be effective in growth promotion. This result showed that $Kiwoomi^{TM}$-treated elementary students significantly increased their growth rate (about 2.14 times), as compared to untreated children. Taken together, it is suggested that this functional rice ($Kiwoomi^{TM}$) might be helpful for growing children without any side effects.