• Title/Summary/Keyword: β-Glucosidase

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Bioconversion of Ginsenosides by Bifidobacterium CBT BG7, BR3 and BL3 (비피도박테리움 CBT BG7, BR3, BL3의 진세노사이드 전환능)

  • Jiwon Choi;Chang Kwon;Jong Won Kim;Myung Jun Chung;Jong Hyun Yoon;Sanghyun Lim
    • Microbiology and Biotechnology Letters
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    • v.50 no.3
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    • pp.395-403
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    • 2022
  • In this study, we identified that the fermentation of Korean indigenous probiotics and red ginseng produced ginsenoside compound K (CK) from major ginsenosides. Based on whole genome sequencing of 19 probiotics species, β-glucosidase, α-arabinofuranosidase, β-xylosidase, and α-rhamnosidase related to bioconversion of ginsenosides are identified in the genome of 19 species, 3 species, 6 species, and 8 species, respectively. Among the 19 probiotics species, Bifidobacterium longum CBT BG7 converted from ginsenoside Rb1 to CK, and both B. breve CBT BR3 and B. lactis CBT BL3 converted ginsenoside Rb1 to Rd. The final concentration and yield of ginsenoside F2 and CK were higher in the fermentation with the nondisrupted cells than with disrupted cells. The combination of both CBT BG7 and BL3, and CBT BG7 and BR3 showed higher amounts of F2 than CBT BG7 only. CBT BG7 with adding α-amylase increased the amounts of F2. In this study, we identified that the fermentation of both Korean indigenous probiotic bacteria CBT BG7, BR3 and BL3, and red gingseng is able to produce CK, a bioactive compound that promotes health benefits.

Effects of a Hwanggi-tang Ethanol Extract on Glucose Uptake and Metabolism in Murine Myotubes (근육세포주에서 당 흡수 및 대사 조절에 대한 황기탕 에탄올 추출물의 효과)

  • Jang, Chul-yong;Shin, Sun-ho;Shin, Yong-jeen
    • The Journal of Internal Korean Medicine
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    • v.41 no.4
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    • pp.599-611
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    • 2020
  • Objectives: The aim of this study was to evaluate the effects of Hwanggi-tang on glucose digestion, uptake, and metabolism in murine C2C12 myotubes. Methods: Hwanggi-tang was prepared according to the Dong-ui-bo-gam (≪東醫寶鑑≫) prescription by 70% ethanol extraction. The effect on glucose digestion was examined by determining the inhibitory effect of Hwanggi-tang on α-glucosidase activity. We also compared and verified the gene and protein expression of genes related to glucose uptake in C2C12 myotubes treated with Hwanggi-tang or insulin. Glucose metabolism was assessed by the expression levels of associated enzymes. Results: Hwanggi-tang caused a dose-dependent inhibition of α-glucosidase activity, induced glucose uptake by activation of the PI3K/Akt/mTOR pathway in the insulin signaling pathway, and promoted glucose oxidation and β-oxidation. Conclusions: Hwanggi-tang exerts an anti-diabetic effect on murine myotubes by inhibiting glucose digestion and inducing glucose uptake and consumption.

Characteristics of Enzymatic Hydrolysis of Sodium Hydroxide pretreated Suwon Poplar (NaOH 전처리된 현사시나무의 효소가수분해 특성)

  • 박영기;오정수
    • Journal of Korea Foresty Energy
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    • v.20 no.2
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    • pp.20-27
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    • 2001
  • An effective method for production of glucose was developed using enzymatic hydrolysis of Suwon poplar by the cellulase. Enzymatic hydrolysis of wood is the reaction to produce glucose from wood using enzyme which derives from microorganism. Glucose can be transferred easily to ethanol by fermentation. Ethanol is the starting material for producing acetone, butanol, citric acid and lactic acid. The mechanism of the enzymatic hydrolysis of cellulose are reasonably explained in terms of the sequential action of three different types of enzymes, endo-cellulase, ex-cellulase, and $\beta$ -glucosidase. The goal of this work was to investigate the cellulose hydrolysis pretreated polar with various concentration NaOH, the crystallinity of cellulose, lignin contents and the degree of hydrolysis.

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Characteristics of Microbial Community Enzyme Activity and Substrate Availability of Damaged Soil (훼손 토양의 미생물군집 효소 활성과 기질 이용성 특성)

  • Ji Seul Kim;Gyo-Cheol Jeong;Myoung Hyeon Cho;Eun Young Lee
    • Journal of Soil and Groundwater Environment
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    • v.28 no.5
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    • pp.68-77
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    • 2023
  • The effect of soil damage on the physicochemical characteristics and activity of the soil microbial community is not well known. This study investigates this relationship by analyzing 11 soil samples collected from various points of soil damage across Gyeonggi-do. Soil damage resulted from forest fires, landslides, and development areas, with their impacts most severe on the topsoil layer (0-30 cm). Dehydrogenase and β-glucosidase activities were notably higher at locations damaged by forest fires compared to other sites. While enzyme activities in soils influenced by landslides and development areas were relatively low, sites with a pollution history exhibited elevated dehydrogenase activity, likely due to past microbial response to the pollution. Additionally, an assessment of carbon substrate usability by soil microorganisms indicated higher substrate availability in areas impacted by forest fires, contrasting with lower availability in landslide and development sites. Statistical analysis revealed a positive correlation between organic content of sand and clay and microbial activity. These findings provide valuable insights into soil damage and associated restoration research, as well as management strategies.

Metagenomic Insight into Lignocellulose Degradation of the Thermophilic Microbial Consortium TMC7

  • Wang, Yi;Wang, Chen;Chen, Yonglun;Chen, Beibei;Guo, Peng;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1123-1133
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    • 2021
  • Biodegradation is the key process involved in natural lignocellulose biotransformation and utilization. Microbial consortia represent promising candidates for applications in lignocellulose conversion strategies for biofuel production; however, cooperation among the enzymes and the labor division of microbes in the microbial consortia remains unclear. In this study, metagenomic analysis was performed to reveal the community structure and extremozyme systems of a lignocellulolytic microbial consortium, TMC7. The taxonomic affiliation of TMC7 metagenome included members of the genera Ruminiclostridium (42.85%), Thermoanaerobacterium (18.41%), Geobacillus (10.44%), unclassified_f__Bacillaceae (7.48%), Aeribacillus (2.65%), Symbiobacterium (2.47%), Desulfotomaculum (2.33%), Caldibacillus (1.56%), Clostridium (1.26%), and others (10.55%). The carbohydrate-active enzyme annotation revealed that TMC7 encoded a broad array of enzymes responsible for cellulose and hemicellulose degradation. Ten glycoside hydrolases (GHs) endoglucanase, 4 GHs exoglucanase, and 6 GHs β-glucosidase were identified for cellulose degradation; 6 GHs endo-β-1,4-xylanase, 9 GHs β-xylosidase, and 3 GHs β-mannanase were identified for degradation of the hemicellulose main chain; 6 GHs arabinofuranosidase, 2 GHs α-mannosidase, 11 GHs galactosidase, 3 GHs α-rhamnosidase, and 4 GHs α-fucosidase were identified as xylan debranching enzymes. Furthermore, by introducing a factor named as the contribution coefficient, we found that Ruminiclostridium and Thermoanaerobacterium may be the dominant contributors, whereas Symbiobacterium and Desulfotomaculum may serve as "sugar cheaters" in lignocellulose degradation by TMC7. Our findings provide mechanistic profiles of an array of enzymes that degrade complex lignocellulosic biomass in the microbial consortium TMC7 and provide a promising approach for studying the potential contribution of microbes in microbial consortia.

Antidiabetic Effect of Aurantii Fructus Immaturus in Streptozotocin-induced Diabetes Model of Mice (Streptozotocin 유도 당뇨병 생쥐 모델에서 지각 추출물의 항당뇨 효과)

  • Kyung-Jae Yi;Ji-Sung Im;Ji-Eun Kim;Su-Kyung Lee;Hyun-Joo Kim;Yung-Sun Song
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.37 no.1
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    • pp.1-8
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    • 2023
  • The aim of this study is to evaluate the antidiabetic effect of the water extract of Aurantii fructus immaturus (WAF), in diabetic models using enzyme, cells and mice, and to suggest a putative mechanism explaining its antidiabetic effect. In an enzyme model using the enzyme α-glucosidase, WAF had no significant effect on α-glucosidase, as compared with acarbose, an antidiabetic drug. Nonetheless, WAF was capable of reducing the blood glucose levels during oral sucrose tolerance test and oral glucose tolerance test, implying that there would be other antidiabetic pathways in no relation to inhibition of α-glucosidase. In cell models using RIN-m5f β-cells and L6 myotubes, WAF, at its non-cytotoxic doses, augmented the secretion of insulin in RIN-m5f β-cells stimulated with 5 mM glucose. In addition, it enhanced the cellular uptake of glucose in L6 myotubes stimulated with deprivation of glucose for 12 h. Therefore, it is most likely that WAF may exert its antidiabetic effects, at least in part, by enhancing insulin secretion and glucose uptake. Meanwhile, in diabetic mice induced with peritoneal injection of streptozotocin (STZ), WAF significantly improved fast blood glucose levels, glycosylated hemoglobin levels, body weight loose, blood pressure, and diabetic adverse effects on functions of the kidney and the liver. Taken together, the water extract of Aurantii fructus immaturus may ameliorate diabetes in mice injected with STZ, at least in part, by enhancing insulin secretion and glucose uptake.

Highly Selective Production of Compound K from Ginsenoside Rd by Hydrolyzing Glucose at C-3 Glycoside Using β-Glucosidase of Bifidobacterium breve ATCC 15700

  • Zhang, Ru;Huang, Xue-Mei;Yan, Hui-Juan;Liu, Xin-Yi;Zhou, Qi;Luo, Zhi-Yong;Tan, Xiao-Ning;Zhang, Bian-Ling
    • Journal of Microbiology and Biotechnology
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    • v.29 no.3
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    • pp.410-418
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    • 2019
  • To investigate a novel ${\beta}$-glucosidase from Bifidobacterium breve ATCC 15700 (BbBgl) to produce compound K (CK) via ginsenoside $F_2$ by highly selective and efficient hydrolysis of the C-3 glycoside from ginsenoside Rd, the BbBgl gene was cloned and expressed in E. coli BL21. The recombinant BbBgl was purified by Ni-NTA magnetic beads to obtain an enzyme with specific activity of 37 U/mg protein using pNP-Glc as substrate. The enzyme activity was optimized at pH 5.0, $35^{\circ}C$, 2 or 6 U/ml, and its activity was enhanced by $Mn^{2+}$ significantly. Under the optimal conditions, the half-life of the BbBgl is 180 h, much longer than the characterized ${\beta}$-glycosidases, and the $K_m$ and $V_{max}$ values are 2.7 mM and $39.8{\mu}mol/mg/min$ for ginsenoside Rd. Moreover, the enzyme exhibits strong tolerance against high substrate concentration (up to 40 g/l ginsenoside Rd) with a molar biotransformation rate of 96% within 12 h. The good enzymatic properties and gram-scale conversion capacity of BbBgl provide an attractive method for large-scale production of rare ginsenoside CK using a single enzyme or a combination of enzymes.

Bacterial endophytes from ginseng and their biotechnological application

  • Chu, Luan Luong;Bae, Hanhong
    • Journal of Ginseng Research
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    • v.46 no.1
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    • pp.1-10
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    • 2022
  • Ginseng has been well-known as a medicinal plant for thousands of years. Bacterial endophytes ubiquitously colonize the inside tissues of ginseng without any disease symptoms. The identification of bacterial endophytes is conducted through either the internal transcribed spacer region combined with ribosomal sequences or metagenomics. Bacterial endophyte communities differ in their diversity and composition profile, depending on the geographical location, cultivation condition, and tissue, age, and species of ginseng. Bacterial endophytes have a significant effect on the growth of ginseng through indole-3-acetic acid (IAA) and siderophore production, phosphate solubilization, and nitrogen fixation. Moreover, bacterial endophytes can protect ginseng by acting as biocontrol agents. Interestingly, bacterial endophytes isolated from Panax species have the potential to produce ginsenosides and bioactive metabolites, which can be used in the production of food and medicine. The ability of bacterial endophytes to transform major ginsenosides into minor ginsenosides using β-glucosidase is gaining increasing attention as a promising biotechnology. Recently, metabolic engineering has accelerated the possibilities for potential applications of bacterial endophytes in producing beneficial secondary metabolites.

Characteristics of Leuconostoc spp. isolated from radish kimchi and its immune enhancement effect (무김치에서 분리한 Leuconostoc 속의 특성과 면역증강 효과)

  • Seoyeon Kwak;Seongeui Yoo;Jieon Park;Woosoo Jeong;Hee-Min Gwon;Soo-Hwan Yeo;So-Young Kim
    • Food Science and Preservation
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    • v.30 no.6
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    • pp.1082-1094
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    • 2023
  • The purpose of this study was to examine the characteristics of Leuconostoc spp. isolated from radish kimchi and to investigate the potential for the use of functional ingredients by evaluating enzymatic characteristics, safety, and immune-enhancing effects among the isolates, including Lactobacillus rhamnosus ATCC53103 (LGG) as a control strain. All test strains exhibited β-glucosidase enzyme activity that releases β-1,4 sugar chain bonds. In addition, as a result of antibiotic resistance assay among the isolates, MIC values on 8 antibiotics were below compared to the EFSA standard, and hemolytic experiments confirmed that all showed gamma hemolysis without hemolytic ability. As a result of the antibacterial activity experiment, the Leu. mesenteroides K2-4 strain showed a higher activity than LGG against Bacillus cereus and Staphylococcus aureus. Additionally, the activity of the NF-kB/AP-1 transcription factor increased when the isolates were treated in macrophage RAW cells. These results were related to increasing the high mRNA expression levels on TNF-α and IL-6 by Leu. mesenteroides K2-4 strain to be treated at low concentration. Consequently, we suggest that it will be useful as a candidate for functional food ingredients.

Antimicrobial, Antioxidant, and Anti-diabetic Activities of Rodgersia podophylla (도깨비부채의 항균, 항산화 및 항당뇨 활성)

  • Pyo, Su-Jin;Lee, Yun-Jin;Kang, Deok-Gyeong;Son, Ho-Jun;Park, Gwang Hun;Park, Jong-Yi;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.30 no.3
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    • pp.298-303
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    • 2020
  • This study aimed to investigate possible applications of Rodgersia podophylla in the food and cosmetic industry. Ethanol extracts of leaves (RP-L), branches (RP-B), and root (RP-R) were prepared, and their antimicrobial, antioxidant, and anti-diabetic activities were evaluated. The polyphenol content in the RP-R, RP-L, and RP-B extracts was 79.6, 30.4, and 16.9 mg/g, respectively. An antimicrobial activity assay showed that the RP-L and RP-R extracts exhibited strong growth inhibition of pathogenic and food spoilage Gram-positive bacteria. Furthermore, the RP-R extract inhibited the growth of the Gramnegative E. coli and P. vulgaris bacteria. All extracts showed strong scavenging activity for DPPH, ABTS, nitrite, and reducing power determined by A 700 nm. In particular, the RC50s of the RP-R extract for the DPPH anion and ABTS cation were 23.0-29.7 and 15.0-18.2 ㎍/ml, respectively, which are comparable to those of vitamin C (9.8 and 8.0 ㎍/ml, respectively). An activity assay of α-glucosidase and β-amylase suggested a high potential for the RP-R extract as an anti-diabetic agent. Its inhibition levels of α-glucosidase and β-amylase at 0.5 mg/ml were 6.9 and 48.5%, respectively. This is the first report of the antimicrobial and anti-diabetic activities of R. podophylla. Our results suggest that RP-L and RP-R extracts could be developed as novel cosmeceutical and functional food resources.