• Title/Summary/Keyword: BGL

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Characterization of Three Extracellular β-Glucosidases Produced by a Fungal Isolate Aspergillus sp. YDJ14 and Their Hydrolyzing Activity for a Flavone Glycoside

  • Oh, Jong Min;Lee, Jae Pil;Baek, Seung Cheol;Jo, Yang Do;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.757-764
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    • 2018
  • A cellulolytic fungus, YDJ14, was isolated from compost and identified as an Aspergillus sp. strain. Three extracellular ${\beta}$-glucosidases, BGL-A1, BGL-A2, and BGL-A3, were separated using ultrafiltration, ammonium sulfate fractionation, and High-Q chromatography. The molecular masses of the three enzymes were estimated to be 100, 45, and 40 kDa, respectively, by SDS-PAGE. The optimum pH and temperature of BGL-A3 were 5.0 and $50^{\circ}C$, respectively, whereas the optimum pH and temperature of BGL-A1 and BGL-A2 were identical (4.0 and $60^{\circ}C$, respectively). The half-life of BGL-A3 at $70^{\circ}C$ (2.8 min) was shorter than that of BGL-A1 and BGL-A2 (12.1 and 8.8 min, respectively). All three enzymes preferred p-nitrophenyl-${\beta}$-$\text\tiny{D}$-glucopyranoside (pNPG) and hardly hydrolyzed cellobiose, suggesting that these enzymes were aryl ${\beta}$-glucosidases. The $K_m$ of BGL-A3 (1.26 mM) for pNPG was much higher than that of BGL-A1 and BGL-A2 (0.25 and 0.27 mM, respectively). These results suggested that BGL-A1 and BGL-A2 were similar in their enzymatic properties, whereas BGL-A3 differed from the two enzymes. When tilianin (a flavone glycoside of acacetin) was reacted with the three enzymes, the inhibitory activity for monoamine oxidase, a target in the treatment of neurological disorders, was similar to that shown by acacetin. We conclude that these enzymes may be useful in the hydrolysis of flavone glycosides to improve their inhibitory activities.

Complete Biotransformation of Protopanaxatriol-Type Ginsenosides in Panax ginseng Leaf Extract to Aglycon Protopanaxatriol by β-Glycosidases from Dictyoglomus turgidum and Pyrococcus furiosus

  • Yang, Eun-Joo;Shin, Kyung-Chul;Lee, Dae Young;Oh, Deok-Kun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.255-261
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    • 2018
  • Aglycon protopanaxatriol (APPT) has valuable pharmacological effects such as memory enhancement and tumor inhibition. ${\beta}$-Glycosidase from the hyperthermophilic bacterium Dictyoglomus turgidum (DT-bgl) hydrolyzes the glucose residues linked to APPT, but not other glycoside residues. ${\beta}$-Glycosidase from the hyperthermophilic bacterium Pyrococcus furiosus (PF-bgl) hydrolyzes the outer sugar at C-6 but not the inner glucose at C-6 or the glucose at C-20. Thus, the combined use of DT-bgl and PF-bgl is expected to increase the biotransformation of PPT-type ginsenosides to APPT. We optimized the ratio of PF-bgl to DT-bgl, the concentrations of substrate and enzyme, and the reaction time to increase the biotransformation of ginsenoside Re and PPT-type ginsenosides in Panax ginseng leaf extract to APPT. DT-bgl combined with PF-bgl converted 1.0 mg/ml PPT-type ginsenosides in ginseng leaf extract to 0.58 mg/ml APPT without other ginsenosides, with a molar conversion of 100%. We achieved the complete biotransformation of ginsenoside Re and PPT-type ginsenosides in ginseng leaf extract to APPT by the combined use of two ${\beta}$-glycosidases, suggesting that discarded ginseng leaves can be used as a source of the valuable ginsenoside APPT. To the best of our knowledge, this is the first quantitative production of APPT using ginsenoside Re, and we report the highest concentration and productivity of APPT from ginseng extract to date.

Nucleotide Sequence of Cellulolytic Xylanase Gene (bglBC2) from Bacillus circulans (Bacillus circulans 유래 cellulolytic xylanase 유전자(bglBC2)의 염기서열 결정 및 분석)

  • Kim, Ji-Yeon
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.67-72
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    • 2006
  • The nucleotide sequence of the cloned cellulolytic xylanase gene (bglBC2) from B. circulans ATCC21367 was determined. bglBC2 consists of an 1,224 bp open reading frame (ORF) coding for a polypeptide of 407 amino acids with a deduced molecular weight of 45 kDa. The Shine-Dalgarno (SD) sequence (5'-AAAGGAG-3') was found 9 bp upstream of the initiation codon, ATG. A promoter region corresponding closely to the B. subtilis consensus sequence (-35: TTGACA,-10: TATAAT) was detected, the putative -35 and -10 sequences of which were TTTACA and TATACT, respectively. The deduced amino acid sequence of the cellulolytic xylanase showed 97% homology with that of the alkaline $endo-\beta-1,4-glucanase$ from B. circulans KSM-N257, 75% homology with that of the $endo-\beta-1,3-1,4-glucanase$ from B. circulans WL-12, and 45% homology with that of the $endo-\beta-1,4-glucanase$ (cellulase) from Bacillus sp. KSM-330. The bglBC2 sequence was deposited in Gen-Bank under the accession number AY269256.

Nutritional and antioxidative properties of black goat meat cuts

  • Kim, Hye-Jin;Kim, Hee-Jin;Jang, Aera
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.9
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    • pp.1423-1429
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    • 2019
  • Objective: In this study, we evaluated the nutritional value and antioxidant activity of black goat loin (BGL) and black goat rump (BGR) meat. Methods: We evaluated the proximate compositions, collagen and mineral contents, and fatty acid compositions of BGL and BGR with respect to their nutritional value. The levels of bioactive compounds such as L-carnitine, creatine, creatinine, carnosine, and anserine were also measured. The ferric reducing antioxidant power (FRAP), 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging, and oxygen radical absorption capacity (ORAC) were assessed to evaluate the antioxidant activity of BGL and BGR. Results: BGR showed higher collagen, Fe, Ca, P, and Na contents than did BGL (p<0.05). Notably, the Ca/P ratio was high in both BGR and BGL (1.82 and 1.54, respectively), thus satisfying the recommendation that the Ca/P ratio is between 1 and 2. BGL showed a significantly higher content of desirable fatty acids (stearic acid and total unsaturated fatty acids) than did BGR. In addition, the levels of creatine, carnosine, and anserine in BGL were higher than those in BGR (p<0.05). There was no significant difference in the antioxidant activity between BGL and BGR, as assessed by FRAP (both $15.92{\mu}mol$ Trolox equivalent [TE]/g of dry matter [DM]), ABTS (12.51 and $12.90{\mu}mol\;TE/g\;DM$, respectively), and ORAC (101.25 and $99.06{\mu}mol\;TE/g\;DM$, respectively) assays. Conclusion: This was a primary study conducted to evaluate the differences in nutritional value and antioxidant activity between loin and rump cuts of black goat meat. Our results provide fundamental knowledge that can help understand the properties of black goat meat.

Comparison in Restriction Profile Analysis of Vibrio furnissi, Vibrio fluvialis, and Vibrio parahaemolyicus Bacteriophage from Sea Product

  • Younghee Kim
    • Journal of Environmental Science International
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    • v.1 no.2
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    • pp.99-103
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    • 1992
  • The bacteriophages lytic for Vibrio furnissi, Vibrio furniulis and Vibrio parahemolyticus were isolated from fish gills and shellfish. Nucleic acid of bacteriophage was prepared and restriction endonuclease profile was compared. All isolates contained deoxyribonucleic acid. V. fumissi bacteriophage from fish gills showed 2 bands with Bgl II, 1 with Pst, 3 with Hind III, 1 with Bm HI and 2 with EcoR I. V Puuialis phage represented 7 fragments with Bgl II, 1 with Pst, 4 with Hind III, and 2 with EcoR I. V parhemolyticn produced 13 sites with Hind III and 4 sites with EcoR I. The fragment types were varied depending on the phage isolation. All three phages were digested with Hind III and EcoR I with different sizes. V furnissi phage were digested with 5 different restriction enzymes. Key words: Bacteriophage, Vibrio furnissi, Vibrio fluvialis, Vibrio pnrahemolyticus, Deoxyribonucleic acid, Pst, Bam HI, Hind III, EcoR I, Bgl II.

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Screening and Characterization of an Enzyme with ${\beta}-Glucosidase$ Activity from Environmental DNA

  • Kim, Soo-Jin;Lee, Chang-Muk;Kim, Min-Young;Yeo, Yun-Soo;Yoon, Sang-Hong;Kang, Han-Cheol;Koo, Bon-Sung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.905-912
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    • 2007
  • A novel ${\beta}-glucosidase$ gene, bglA, was isolated from uncultured soil bacteria and characterized. Using genomic libraries constructed from soil DNA, a gene encoding a protein that hydrolyzes a fluorogenic analog of cellulose, 4-methylumbelliferyl ${\beta}-D-cellobioside$ (MUC), was isolated using a microtiter plate assay. The gene, bglA, was sequenced using a shotgun approach, and expressed in E. coli. The deduced 55-kDa amino acid sequence for bglA showed a 56% identity with the family 1 glycosyl hydrolase Chloroflexus aurantiacus. BglA included two conserved family 1 glycosyl hydrolase regions. When using $p-nitrophenyl-{\beta}-D-glucoside$ (pNPG) as the substrate, the maximum activity of the purified ${\beta}-glucosidase$ exhibited at pH 6.5 and $55^{\circ}C$, and was enhanced in the presence of $Mn^{2+}$. The $K_m\;and\;V_{max}$ values for the purified enzyme with pNPG were 0.16 mM and $19.10{\mu}mol/min$, respectively. The purified BglA enzyme hydrolyzed both pNPG and $p-nitrophenyl-{\beta}-D-fucoside$. The enzyme also exhibited substantial glycosyl hydrolase activities with natural glycosyl substrates, such as sophorose, cellobiose, cellotriose, cellotetraose, and cellopentaose, yet low hydrolytic activities with gentiobiose, salicin, and arbutin. Moreover, BglA was able to convert the major ginsenoside $Rb_1$ into the pharmaceutically active minor ginsenoside Rd within 24 h.

Gene Cloning of Cellulose Degradation Enzyme of Bacillus subtilis LYH201 Strain (Bacillus subtilis LYH201균주의 섬유소 분해효소의 유전자 Cloning 및 특성분석)

  • Lee, Young-Han;Park, Sang-Ryeol
    • Korean Journal of Soil Science and Fertilizer
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    • v.34 no.5
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    • pp.333-341
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    • 2001
  • The Compost-decomposing-bacteria was isolated from livestock compost containing sawdust. The isolated bacteria was identified as Bacillus subtilis LYH201 by the method of the composition of the fatty acid with MIDI system and Bergey's manual. Cloning of CMCase encoding gene was accompanied by shotgun method. The pLK100 have yellow activity ring on CMC medium, that was carried 2.2 kb insert DNA in pBluescript II $SK^+$ vector, named BglC gene. The BglC was very similar to Pectobacterium carotovorum Gun_CLOAB(P15704) with score of 57% identity and 71% homology over 508 aa. The BglC was measured molecular weight 56 kDa by CMC-SDS-PAGE. Optimum cellulase activity Bacillus subtilis LYH201 was temperature $50^{\circ}C$ and pH 7.

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Comparison of Blood Glucose Level in Pancreatic Cancer on $^{18}F$-FDG PET (췌장암 환자의 양전자방출단층촬영(PET) 검사 시 혈당치와 영상에 관한 분석)

  • Oh, Shin-Hyun;Lee, Seung-Jae;Park, Hoon-Hee;Park, Yong-Sung;Kang, Chun-Goo;Lim, Han-Sang;Kim, Jae-Sam;Lee, Chang-Ho
    • The Korean Journal of Nuclear Medicine Technology
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    • v.15 no.1
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    • pp.65-69
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    • 2011
  • Purpose: Normal pancreas has low uptake rate in $^{18}F$-FDG PET scan. However, it is possible to diagnose malignancy of pancreatic cancer which has high uptake rate. Many studies approve a high prevalence of diabetes in pancreatic cancer and if the blood glucose level (BGL) is over the normal range, FDG uptake will be decreased and there will be inconvenience for patients from the delay time to reduce the high BGL or could cause difficulty to arrange the schedule. Therefore, we studied the relation of BGL and image quality in pancreatic cancer on PET. Materials and Methods: A hundred patients had PET scan. The prevalence of pancreatic cancer and diabetes were evaluated using SPSS ver. 17. The fasting BGL of patients were examined and sorted as diabetes mellitus (DM) group and Non-DM group. For the evaluation, patients were divided into 3 groups (Non-DM, DM; BGL${\geq}7.0mmol/L$, and DM; BGL<7.0 mmol/L). The ROI was drown on Liver and Lung for the PET imaging analysis. Results: Fifty three male and forty seven female were in the patients. The average age was $60.1{\pm}13.5$. There were 36 patients (male: 22, female: 14, 36%) who had pancreatic cancer with DM. There were 15 patients who showed over 7.0 mmol/L in their fasting BGL and 85 patients who showed under 7.0 mmol/L in their fasting BGL. Among the Non-DM, DM; BGL${\geq}7.0mmol/L$, and DM; BGL<7.0 mmol/L, there was not a statistical significance (p>0.05). Conclusion: The prevalence of pancreatic cancer was comparatively high in this study. If the fasting BGL was slightly over the normal BGL, we believe it will not give a severe disturbance when the patients have PET scan. Furthermore, the examination schedule doesn't need to change and the inconvenience from the delay of patients preparation will be reduced.

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A Study of Diabetic Control for Diabetic Patient in $^{18}F$-FDG PET ($^{18}F$-FDG PET 검사를 위한 당뇨 환자의 혈당 조절에 관한 연구)

  • Cha, Min-Kyeong;Nam, Ki-Pyo;Lee, Young-Hee;Choi, Jae-Min;Cho, Shee-Man
    • The Korean Journal of Nuclear Medicine Technology
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    • v.12 no.1
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    • pp.57-61
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    • 2008
  • Purpose: $^{18}F$-FDG PET is to evaluate the tumor using glucose metabolism. Blood Glucose Level (BGL) is important factor that affects on a result of examination. But it often appears the distort result on diabetic patient due to a failure of BGL control. The aim of this study is to make a effective guideline for diabetic patient prescribed $^{18}F$-FDG PET and improve accuracy and reliability of examination. Materials and methods: A subject of study is 69 diabetic patients and divided them into 3 groups; 1) BGL is less than 120 mg/dl & performed PET, 2) BGL is over than 120 mg/dl & performed PET, 3) BGL is over than 120 mg/dl & not performed PET. And we investigated the type of diabetic medications and the time of dosage, whether they have meal or not. And we analyzed SUV of liver and mediastinum to recognize the effect of BGL on PET images. Results: As a methods to diabetic control, Amalyl showed high percentage of 27.5% in oral medication and Humulin R showed 14.5% in insulin. Their peak time was 2-3 hrs, 2-4 hrs and duration time was 24 hrs, 5-7 hrs. The number of that had a breakfast was 16, 7 mens (43.8%) couldn't perform PET as over 120 mg/dl and 5 mens (31.3%) performed PET as less than 120 mg/dl after they took a medication. When we set the SUV of normal for 1.5-2.0, 1.0-1.5 on liver and mediastinum, the percentage out of the range was 31.8%, 52.9%, 62.5% in 120-139 mg/dl, 140-159 mg/dl, 160-200 mg/dl respectively. Conclusions: We completed standardized guideline for diabetic patients. As a result of this study, improved customer satisfaction and increased economic benefits. It is expect to be a effective model in other PET centers.

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Optimization of ${\beta}$-Glucosidase Production by a Strain of Stereum hirsutum and Its Application in Enzymatic Saccharification

  • Ramachandran, Priyadharshini;Nguyen, Ngoc-Phuong-Thao;Choi, Joon-Ho;Kang, Yun Chan;Jeya, Marimuthu;Lee, Jung-Kul
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.351-356
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    • 2013
  • A high ${\beta}$-glucosidase (BGL)-producing strain, Stereum hirsutum, was identified and isolated and showed a maximum BGL activity (10.4 U/ml) when cultured with Avicel and tryptone as the carbon and nitrogen sources, respectively. In comparison with other BGLs, BGL obtained from S. hirsutum showed a higher level of activity to cellobiose ($V_{max}$ = 172 U/mg, and $k_{cat}$ = 281/s). Under the optimum conditions (600 rpm, $30^{\circ}C$, and pH 6.0), the maximum BGL activity of 10.4 U/ml with the overall productivity of 74.5 U/l/h was observed. BGL production was scaled up from a laboratory scale (7-L fermenter) to a pilot scale (70-L fermenter). When S. hirsutum was cultured in fed-batch culture with rice straw as the carbon source in a 70-L fermenter, a comparable productivity of 78.6 U/l/h was obtained. Furthermore, S. hirsutum showed high levels of activity of other lignocellulases (cellobiohydrolase, endoglucanase, xylanase, and laccase) that are involved in the saccharification of biomasses. Application of S. hirsutum lignocellulases in the hydrolysis of Pinus densiflora and Catalpa ovata showed saccharification yields of 49.7% and 43.0%, respectively, which were higher than the yield obtained using commercial enzymes.