• Title/Summary/Keyword: GH16

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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.

Characterization of Exolytic GH50A β-Agarase and GH117A α-NABH Involved in Agarose Saccharification of Cellvibrio sp. KY-GH-1 and Possible Application to Mass Production of NA2 and L-AHG (Cellvibrio sp. KY-GH-1의 아가로오스 당화 관련 엑소형 GH50A β-아가레이즈와 GH117A α-NABH의 특성 및 NA2와 L-AHG 양산에의 적용 가능성)

  • Jang, Won Young;Lee, Hee Kyoung;Kim, Young Ho
    • Journal of Life Science
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    • v.31 no.3
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    • pp.356-365
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    • 2021
  • Recently, we sequenced the entire genome of a freshwater agar-degrading bacterium Cellvibrio sp. KY-GH-1 (KCTC13629BP) to explore genetic information encoding agarases that hydrolyze agarose into monomers 3,6-anhydro-L-galactose (L-AHG) and D-galactose. The KY-GH-1 strain appeared to possess nine β-agarase genes and two α-neoagarobiose hydrolase (α-NABH) genes in a 77-kb agarase gene cluster. Based on these genetic information, the KY-GH-1 strain-caused agarose degradation into L-AHG and D-galactose was predicted to be initiated by both endolytic GH16 and GH86 β-agarases to generate NAOS (NA4/NA6/NA8), and further processed by exolytic GH50 β-agarases to generate NA2, and then terminated by GH117 α-NABHs which degrade NA2 into L-AHG and D-galactose. More recently, by employing E. coli expression system with pET-30a vector we obtained three recombinant His-tagged GH50 family β-agarases (GH50A, GH50B, and GH50C) derived from Cellvibrio sp. KY-GH-1 to compare their enzymatic properties. GH50A β-agarase turned out to have the highest exolytic β-agarase activity among the three GH50 isozymes, catalyzing efficient NA2 production from the substrate (agarose, NAOS or AOS). Additionally, we determined that GH117A α-NABH, but not GH117B α-NABH, could potently degrade NA2 into L-AHG and D-galactose. Sequentially, we examined the enzymatic characteristics of GH50A β-agarase and GH117A α-NABH, and assessed their efficiency for NA2 production from agarose and for production of L-AHG and D-galactose from NA2, respectively. In this review, we describe the benefits of recombinant GH50A β-agarase and GH117A α-NABH originated from Cellvibrio sp. KY-GH-1, which may be useful for the enzymatic hydrolysis of agarose for mass production of L-AHG and D-galactose.

Effect of Exogenous Porcine GH Administration on GH Responses to GH-releasing Peptide-2 and GH-releasing Hormone in Swine

  • Nou, V.;Tomoshi, K.;Inoue, H.;Matsunaga, N.;Kuwayama, H.;Hidari, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.8
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    • pp.1188-1192
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    • 2003
  • Negative feedback on GH responses to GH-releasing hormone (GHRH) and GH-releasing peptides (GHRPs) has been reported and this action has been suggested to act through an increase in somatostatin. To determine whether the acute administration of porcine GH (pGH) inhibits GH responsiveness to GHRP-2 and GHRH in swine, swine were intravenously administered with pGH (5${\mu}g$/kg BW) or placebo followed 180 min later by a second intravenous administration of saline, GHRP-2 (30 ${\mu}g$/kg BW), GHRH (1${\mu}g$/kg BW) and a combination of GHRP-2 and GHRH. Plasma GH concentration was measured by radioimmunoassay. Administration of pGH caused a significant increase in GH area under curve and GH peak concentrations (p<0.001) over placebo-treated group. Plasma GH concentrations peaked at 15 min and returned to baseline level within 90 min. Pretreatment of pGH abolished (p<0.01) GH response to GHRH and attenuated (p<0.05) GH response to GHRP-2 and GHRH combined, without affecting GH response to GHRP-2. These results demonstrate that negative feedback action on GH releasing effect of GHRH occurs in swine, and that GHRP-2 has ability to interact in this action.

Effect of Twice Daily Administration of GH-releasing Peptide-2 for 10 Days on Growth Performance, Plasma GH Responses and Insulin-like Growth Factor-1 Concentrations in Swine

  • Nou, V.;Inoue, H.;Lee, H.G.;Matsunaga, N.;Kuwayama, H.;Hidari, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.8
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    • pp.1193-1198
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    • 2003
  • An increase in frequency of administration of exogenous growth hormone (GH) or GH-releasing hormone was reported to be a model to increase blood circulating insulin-like growth factor-1 (IGF-1) and to improve growth performance in animals. We have investigated the effect of twice daily administration of GH-releasing peptide-2 (GHRP-2) on growth performance, GH responsiveness and plasma insulin-like growth factor IGF-1 in swine. We administered to eight swine, 3 control and 5 treatment, a twice daily s.c. injections of GHRP-2 ($30{\mu}g/kg\;BW$) for a period of 10 days. Every day blood samples immediately taken before injections of GHRP-2 or saline, at 08:00 h and 16:00 h, were measured for IGF-1 concentrations. Blood samples for GH assay were collected every 20 min on days 1, 6 and 10, from 1 hour before and 3 h after GHRP-2 or saline injections at 08:00 h. GH peak concentrations and GH area under curve (GH AUC) on day 1, 6 and 10 in treatment group of swine were higher than those in control swine (p<0.05). Twice daily administration of GHRP-2 caused a significantly attenuation (p<0.05) of GH peak concentrations ($80.25{\pm}13.87$, $39.73{\pm}5.72$ and $27.57{\pm}6.06ng/ml$ for day 1, 6 and 10, respectively) and GH AUCs ($3,536.15{\pm}738.35$, $1,310.31{\pm}203.55$ and $934.37{\pm}208.99ng/ml$ for day 1, 6 and 10, respectively). However, there was no significant difference in GH peak concentration and GH AUC between day 6 and 10. Plasma IGF-1 concentration levels were higher in treatment than control group of swine (p<0.05) after 3 days of the treatment, and the levels reached a plateau from day 3 to 10 of experiment. Growth performance did not alter by GHRP-2 administration, even though a numerical increase of body weight gain and feed efficiency was observed. These results indicate that twice daily administration of GHRP-2 for 10 days in swine did not significantly influence on growth performance, caused an overall attenuation of GH response, and that elevation of plasma GH concentrations caused by GHRP-2 administration increased plasma IGF-1 concentrations, even though an attenuation of GH response was observed.

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.

Characterization of Multifunctional Bacillus sp. GH1-13 (복합기능성 Bacillus sp. GH1-13 균주의 특징)

  • Kim, Sang Yoon;Sang, Mee Kyung;Weon, Hang-Yeon;Jeon, Young-Ah;Ryoo, Jae Hwan;Song, Jaekyeong
    • The Korean Journal of Pesticide Science
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    • v.20 no.3
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    • pp.189-196
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    • 2016
  • Several microorganisms in particular Bacillus subtilis group have been isolated from diverse places such as soils and the gastrointestinal tract of ruminants etc., and used as biocontrol agent against various plant pathogens and utilized as plant growth promoting agents. Among them, Bacillus is well known as one of the most useful bacteria for biocontrol and plant growth promotion. Bacterium GH1-13 was isolated from a reclaimed paddy field in Wando Island and identified as Bacillus velezensis using phylogenetic analysis on the basis of 16S rRNA and gyrB gene. It was confirmed that GH1-13 produced indole acetic acid (IAA) associated with promoted growth of rice root. GH1-13 showed characteristics of antagonization against the main pathogen of rice as well as diverse pathogenic fungi. GH1-13 had biosynthetic genes, bacillomycin, bacilycin, fengycin, iturin, and surfactin which are considered to be associated closely with inhibition of growth of pathogenic fungi and bacteria. This study showed that GH1-13 could be used as a multifunctional agent for biocontrol and growth promotion of crop.

Inhibitory Effect of Guanidine Hydrochloride on the Infectious Flacherie Virus of the Silkworm, Bombyx mori (누에 전염성 연화병의 발병 억제제에 의한 방제)

  • 강석권;김근영;이재창;조용섭
    • Journal of Sericultural and Entomological Science
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    • v.25 no.2
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    • pp.37-43
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    • 1984
  • The prevalence of the infectious flacherie virus (FV) disease causes a severe damage to cocoon yield and various methods to control the disease have been studied. In this regard, guanidine hydrochloride (GH), one of the guanidine derivatives known as the most inhibitory agent against the replication of picorna virus, was applied to silkworms per os with mulberry leaves and the results were as follows. 1. The application of GH below 0.01% of the chemical concentration did not give any damage to silkworm larvae. 2. The transmission of the virus disease by introducing the FV infected larvae to the healthy larvae group was proportioned to the number of infected larvae. When l% of infected larvae was introduced to the rearing tray of healthy larvae, the pupation rate was 70.7%(79) and it was 38.4% (43) to 5% of infected larvae introduced, while the control of non-mixed with infected larvae gave 89.2% (100) of pupation rate. The cocoon yield from 10,000 larvae also showed the same tendency as the pupation rate. 3. The inhibitory effect of GH against the replication of FV showed ten times in treatment of 0,01% of the chemical agent compared to the non-treatment. 4. The successive application of GH after virus inoculation to silkworm larvae led to the most effective on the inhibition of the virus replication. 5. The immediate application of GH after the virus inoculation also gave the best effect on the inhibition of the virus replication in silkworm larvae. 6. The effect of GH on the inactivation of FV in vitro was not observed.

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Growth Hormone 1 T1663A Polymorphism, Recreational Physical Activity and BMI, and Breast Cancer Risk in Chinese Women

  • Gao, Chang-Ming;Ding, Jian-Hua;Wu, Jian-Shong;Cao, Hai-Xia;Li, Su-Ping;Liu, Yan-Ting;Tang, Jin-Hai;Tajima, Kazuo
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5421-5425
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    • 2015
  • To evaluate the relationship between the growth hormone 1 (GH1) T1663A polymorphism, recreational physical activity and body mass index (BMI) with reference to breast cancer, we conducted a case-control study with 669 cases of breast cancer and 682 population-based controls in Jiangsu Province, China. A structured questionnaire was used to elicit detailed information. All subjects completed an in-person interview. GH1 genotypes were identified using PCR-RFLP methods. Odds ratios (ORs) were estimated with an unconditional logistic model. The distribution of GH1 genotypes was not significantly different between controls and cases ($x^2$=2.576, P=0.276). Results of stratified analysis by the participation status of the recreational physical activity showed that the persons with GH1 A allele were at a decreased risk of breast cancer (adjusted-OR=0.66; 95% CI, 0.50-0.87) only among inactive individuals. Stratified analysis by BMI showed that the genotype A/A was associated with a decreased risk of breast cancer only among individuals of the BMI <25 (adjusted-OR=0.80; 95% CI, 0.66-0.98). The findings of this study suggest that recreational physical activity and BMI may modify any association between the GH1 T1663A polymorphism and breast cancer risk.

Growth Outcome and Metabolic Profile of PWS Patients Treated with GH and Differences between AGA and SGA Group

  • Yoon, Ju Young
    • Journal of Interdisciplinary Genomics
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    • v.4 no.2
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    • pp.35-38
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    • 2022
  • Background: Prader-Willi syndrome (PWS) is a complex genetic disease associated with growth impairment, severe obesity and metabolic dysfunctions. High proportion of PWS patients are born small for gestational age (SGA) than normal children, which also increase the risk of growth impairment and metabolic dysfunction in PWS. We aimed to compare growth outcome and metabolic profiles between SGA and appropriate for gestational age (AGA) PWS patients. Methods: Data of 55 PWS children and adults aged more than 2 years old (32 male and 23 female, age 2-18.8 years) from single center were studied. Only patients who were treated with GH were included. The clinical characteristics and laboratory findings were reviewed retrospectively. Results: Among 55 subjects, 39 had 15q11-13 deletion and 16 had uniparental disomy (UPD). Twenty (36.3%) were born SGA. All patients received GH treatment, and 11 (20%) discontinued GH treatment. Mean age at GH treatment initiation was 2.5 (range 0.3-12.4) years, and mean duration of treatment was 6.3 (range 1.0-11.3) years. Current height-SDS (-0.36 vs -0.16) and BMI-SDS (1.44 vs 1.33) did not differ between AGA and SGA group. Two patients in SGA group, but none in AGA group had diabetes mellitus. Mean glucose level was also higher in SGA group (100.1 vs 114.4 mg/dL). Conclusion: Our report gives an overview of growth profile and metabolic dysfunctions recorded in GH treated PWS patients. Growth profile did not differ between AGA and SGA group. Glucose level was higher in SGA group, so more careful monitoring and prevention for DM will be required in SGA group.

The Effects of Sunghyangjunggi-san and Gwackhyangjunggi-san Extracts on Cerebral Ischemia Following the MCA Occlusion in Rat (흰쥐의 중대뇌동맥 결찰로 유발된 뇌허혈에서 성향정기산과 곽향정기산이 신경세포에 미치는 효과)

  • 김선영;이원철
    • The Journal of Korean Medicine
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    • v.22 no.4
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    • pp.142-150
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    • 2001
  • Objectives : The purpose of this investigation is to evaluate and compare the effects of Sunghyangjunggi-san (SH) and Gwackhyangjunggi-san (GH) extracts on reperfusion following the MCA occlusion in rats. Methods : To evaluate the effect of Sunghyangjunggi-san (SH) and Gwackhyangjunggi-san (GH) extracts on reperfusion following the MCA occlusion, the volume of cerebral infarction and edema were measured and the change of the CA1 pyramidal neurons in the hippocampus were investigated by light microscopy. Results : 1. The infarction volume of the control group was 23.6%, that of the GH group was 23.7%, and that of the SH group was 18.5%. 2. The brain edema volume of the control group increased by 16% compared with that of the normal group, that of the GH group increased by 14%, and that of the SH group increased by 9%. 3. The number of surviving pyramidal neurons in the CAI area of the hippocampus was investigated under light microscopy. In the control group, few surviving pyramidal neurons excisted (mean 6.4) and similarly in the GH group (mean 8.5), but in the SH group, the number of surviving pyramidal neurons was significantly higher, to the mean 18.4. Conclusions : According to the above results, in regard to the damage of neurons following cerebral ischemia, the GH group has little effect of the protection of neurons compared to the control group, but the SH group has a remarkable effect.

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