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Enhanced pectinase and β-glucosidase enzyme production by a Bacillus subtilis strain under blue light-emitting diodes

  • Elumalai, Punniyakotti;Lim, Jeong-Muk;Oh, Byung-Teak
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.109-109
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    • 2018
  • Bacillus subtilis B22, a chemotrophic and aerobic bacterial strain was isolated from homemade kimchi, identified by 16S rRNA gene sequencing. B22 was primarily screened by biochemical, carbon source utilization tests. B22 was used to produce pectinase and ${\beta}$-glucosidase by submerged fermentation under different light sources. B22 was incubated in pectin media and basal media (pH 7.0) under blue, green, red and white light-emitting diodes (LEDs), fluorescent white light, and in darkness at $37^{\circ}C$, orbital shaker 150 rpm for 24 hours. Fermentation under blue LEDs maximized pectinase production ($71.59{\pm}1.6U/mL$ at 24 h) and ${\beta}$-glucosidase production ($56.31{\pm}1.6U/mL$ at 24 h). Further, the production of enzyme increased to pectinase ($156{\pm}1.28U/mL$) and ${\beta}$-glucosidase ($172{\pm}1.28U/mL$) with 3% glucose as a carbon source. Activity and stability of the partially purified enzymes were higher at pH 6.0 to 8.0 and $25-55^{\circ}C$. The effect on the metal ions $Na^+$ and $K^+$ and (moderateactivity) $Mn^{2+}$ and $Ni^{2+}$ increased activity, while $Hg^{2+}$, $Cu^{2+}$, $Fe^{2+}$, and $Fe^{2+}$ inhibited activity. EDTA, phenylmethylsulfonyl fluoride and 5,5-dithiobis (2-nitrobenzoicacid) reduced activity, while tetrafluoroethylene and 1,10-phenanthroline inhibited activity. The amylase was highly tolerant of the surfactants TritonX-100, Tween-20, Tween-80 and compatible with organic solvents methanol, ethanol, isoamylalcohol, isopropanol, t-butylalcohol and the oxidizing agents hydrogen peroxide, sodium perborate and sodium hypochlorite, although potassium iodide and ammonium persulfate reduced activity. These properties suggest utility of pectinase and ${\beta}$-glucosidase produced by B. subtilis B22 under blue LED-mediated fermentation for industrial applications.

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Biological Activities of Phellinus linteus Mycelium Culture with Cassiae Semen Extract on β-Glucuronidase Inhibitory Activity (β-Glucuronidase 저해 활성이 우수한 결명자를 첨가한 상황 균사체 배양액의 생리활성)

  • Oh, Eun-Hee;Park, Jung-Mi;Kim, Sang-Hee;Song, In-Gyu;Han, Nam-Soo;Yoon, Hyang-Sik
    • The Korean Journal of Food And Nutrition
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    • v.25 no.3
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    • pp.620-628
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    • 2012
  • We examined the effects of biological activity Phellinus linteus mycelium culture with cassiae semen extract. Firstly, the optimal temperature, initial pH and culture period for mycelial growth in a liquid culture of P. linteus were determined, and they were $30^{\circ}C$, pH 5.0 and 8 days respectively. The five herbal materials were examined against several health functional efficacies, and, as a result, Cassiae semen was chosen, with its superior inhibitory effects in ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities(95.3%, 80.9%, 96.1 and 24.2%, respectively). P. linteus fruit body was investigated on ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities, and they were 54.7%, 81.9%, 30.0% and 20.1%, respectively. Accordingly, C. semen was used in the following experiment, to give an additive functional effect on the P. linteus. As the amount of C. semen in the cultural media increased, mycelial weight and ${\beta}$-glucan contents also increased, but final pH was not influenced. In addition, the ${\beta}$-glucuronidase inhibitory activity, electron donating activity, and ${\alpha}$-glucosidase inhibitory activity increased. P. linteus mycelium culture showed higher activities in the other three tests above, except for electron donating activity, when C. semen was added to the medium before cultivation.

Effect of Solid-State Fermented Brown Rice Extracts on 3T3-L1 Adipocyte Differentiation

  • Su Bin Ji;Chae Hun Ra
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.926-933
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    • 2023
  • Aspergillus oryzae KCCM 11372 was used to enhance the production of β-glucan using humidity control strategies. Under conditions of 60% humidity, solid-state fermentation (SSF) increased the yields of enzymes (amylase and protease), fungal biomass (ergosterol), and β-glucan. The maximum concentrations obtained were 14800.58 U/g at 72 h, 1068.14 U/g at 120 h, 1.42 mg/g at 72 h, and 12.0% (w/w) at 72 h, respectively. Moreover, the β-glucan containing fermented brown rice (β-glucan-FBR) extracts at concentrations of 25-300 ㎍/ml was considered noncytotoxic to 3T3-L1 preadipocytes. We then studied the inhibitory effects of the extracts on fat droplet formation in 3T3-L1 cells. As a result, 300 ㎍/ml of β-glucan-FBR extracts showed a high inhibition of 38.88% in lipid accumulation. Further, these extracts inhibited adipogenesis in the 3T3-L1 adipocytes by decreasing the expression of C/EBPα, PPARγ, aP2, and GLUT4 genes.

Bioconversion of nutrient and phytoestrogen constituents during the solid-state fermentation of soybeans by mycelia of Tricholoma matsutake (송이버섯 균사체를 이용한 대두 고체발효 중 영양성분과 식물성 에스트로겐 성분의 생물전환)

  • Hee Yul Lee;Kye Man Cho;Ok Soo Joo
    • Food Science and Preservation
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    • v.30 no.6
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    • pp.1012-1028
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    • 2023
  • The findings of this study confirmed the alteration of β-glucosidase activity, nutritional constituents, isoflavones, antioxidant activities, and digestive enzyme inhibition activities in soybeans during solid-state fermentation times with mycelia of Tricholoma matsutake. After nine days, the highest activity level was observed for β-glucosidase (3.90 to 38.89 unit/g) and aglycones (163.03 to 1,074.28 ㎍/g). The sum of isoflavones showed a significant decrease (3,489.41 to 1,325.66 ㎍/g) along with glycosides (2,753.87 to 212.43 ㎍/g) for fermentation, while fatty acids showed a slight increase and amino acids showed a marked increase. Total phenolic and flavonoid contents showed a corresponding increase according to fermentation times (5.58 to 15.09 GAE mg/g; 0.36 to 1.58 RE mg/g). Antioxidant and enzyme inhibition activities also increased; in particular, the highest level of scavenging activities was observed for ABTS (up 60.13 to 82.08%), followed by DPPH (up 63.92% to 71.98%) and hydroxyl (up 36.01 to 52.02%) radicals. Of particular interest, α-glucosidase (6.69 to 83.49%) and pancreatic lipase inhibition (1.22 to 77.43%) showed a marked increase. These results demonstrated that fermentation of soybeans with the mycelia of T. matsutake enhanced the nutritional and functional constituents, and the biological activities of soybeans. Thus, this fermentation technology can be used to produce a novel functional materials from soybeans.

Changes in Biological Qualities of Soy Grits Cheonggukjang by Fermentation with β-Glucosidase-Producing Bacillus Strains (β-Glucosidase 활성이 있는 균주 Bacillus Strains를 접종해 제조한 Soy Grits 청국장의 품질 특성)

  • Lee, Kyung Ha;Choi, Hye Sun;Hwang, Kyung A;Song, Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.5
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    • pp.702-710
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    • 2016
  • The bioactivity of soy grits cheonggukjang was enhanced by fermentation using three ${\beta}$-glucosidase-producing Bacillus subtilis strains (HJ 18-9, HJ 25-8, and HJ 18-9+HJ 25-8) for 48 h at $37^{\circ}C$. The results indicate that protease, cellulase, and a-amylase activities significantly increased (P<0.05) with increasing fermentation time. In addition, the amino-type nitrogen content of B. subtilis-fermented soy grits cheonggukjang increased to 91.0~168.0 mg% after 48 h of fermentation. Among the isoflavones in soy grits cheonggukjang, contents of ${\beta}$-glucosides or acetyl-glucosides were reduced while aglycone content increased upon fermentation. In particular, soy grits cheonggukjang fermented with B. subtilis HJ18-9 and HJ25-8 showed the largest increases in aglycone content compared to complex treatment. These results provide useful information for development starter (single and complex) as well as for production of high quality fermented soybean food.

Optimization of Glycosyl Aesculin Synthesis by Thermotoga neapolitana β-Glucosidase Using Response-surface Methodology (반응표면분석법을 이용한 Thermotoga neapolitana β-glucosidase의 당전이 활성을 통한 glycosyl aesculin 합성 최적화)

  • Park, Hyunsu;Park, Young-Don;Cha, Jaeho
    • Journal of Life Science
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    • v.27 no.1
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    • pp.38-43
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    • 2017
  • Glycosyl aesculin, a potent anti-inflammatory agent, was synthesized by transglycosylation reaction, catalyzed by Thermotoga neapolitana ${\beta}-glucosidase$, with aesculin as an acceptor. The key reaction parameters were optimized using response-surface methodology (RSM) and $2{\mu}g$ of the enzyme. As shown by a statistical analysis, a second-order polynomial model fitted well to the data (p<0.05). The response surface curve for the interaction between aesculin and other parameters revealed that the aesculin concentration and reaction time were the primary factors that affected the yield of glycosyl aesculin. Among the tested factors, the optimum values for glycosyl aesculin production were as follows: aesculin concentration of 9.5 g/l, temperature of $84^{\circ}C$, reaction time of 81 min, and pH of 8.2. Under these conditions, 61.7% of glycosyl aesculin was obtained, with a predicted yield of 5.86 g/l. The maximum amount of glycosyl aesculin was 6.02 g/l. Good agreement between the predicted and experimental results confirmed the validity of the RSM. The optimization of reaction conditions by RSM resulted in a 1.6-fold increase in the production of glycosyl aesculin as compared to the yield before optimization. These results indicate that RSM can be effectively used for process optimization in the synthesis of a variety of biologically active glycosides using bacterial glycosidases.

Single-dose Oral Toxicity Study of β-glucosidase 1 (AtBG1) Protein Introduced into Genetically Modified Rapeseed (Brassica napus L.) (GM 유채에 도입된 β-glucosidase 1 (AtBG1)의 단회투여독성시험)

  • Lee, Soonbong;Jeong, Kwangju;Jang, Kyung-Min;Kim, Sung-Gun;Park, Jung-Ho;Kim, Shinje
    • Journal of Life Science
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    • v.27 no.2
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    • pp.194-201
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    • 2017
  • Rapeseed (Brassica napus L.) is an oil crop classified as Brassicaceae, and it is widely grown worldwide. To develop a drought-resistant rapeseed, the ${\beta}$-glucosidase 1 (AtBG1) gene was introduced into rapeseed because drought- and salt-resistance phenotypes were observed when the AtBG1 gene was overexpressed in arabidopsis. Newly developed genetically modified crop must be proved to be safe. Safety assessments are based on the historical usage and scientific reports of a crop. In this study, we examined the potential acute oral toxicity of AtBG1 protein expressed in genetically modified (GM) rapeseed and calculated the minimum lethal dose at 6 weeks in both male and female ICR mice. AtBG1 protein was fed at a dose of 2,000 mg/kg body weight in five male and five female mice according to the marginal capacity concentration of OECD, 2,000 mg/15 ml/kg. Mortalities, clinical findings, and body weight changes were monitored for 14 days after dosing, and postmortem necropsy was performed on day 14. This study showed that no deaths occurred in the test group, and AtBG1 protein did not result in variations in common symptoms, body weight, and postmortem findings between the two groups. This showed that the minimum lethal dose of AtBG1 protein expressed in transgenic rapeseed exceed 2,000 mg/kg body weight in both sexes.

Safety Evaluation of Bifidobacterium breve IDCC4401 Isolated from Infant Feces for Use as a Commercial Probiotic

  • Choi, In Young;Kim, Jinhee;Kim, Su-Hyeon;Ban, O-Hyun;Yang, Jungwoo;Park, Mi-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.949-955
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    • 2021
  • Previously, our research group isolated Bifidobacterium breve IDCC4401 from infant feces as a potential probiotic. For this study, we evaluated the safety of B. breve IDCC4401 using genomic and phenotypic analyses. Whole genome sequencing was performed to identify genomic characteristics and investigate the potential presence of genes encoding virulence, antibiotic resistance, and mobile genetic elements. Phenotypic analyses including antibiotic susceptibility, enzyme activity, production of biogenic amines (BAs), and proportion of D-/L-lactate were evaluated using E-test, API ZYM test, high-performance liquid chromatography (HPLC), and D-/L-lactic acid assay respectively. The genome of B. breve IDCC4401 consists of 2,426,499 bp with a GC content of 58.70% and 2,016 coding regions. Confirmation of the genome as B. breve was provided by its 98.93% similarity with B. breve DSM20213. Furthermore, B. breve IDCC4401 genes encoding virulence and antibiotic resistance were not identified. Although B. breve IDCC4401 showed antibiotic resistance against vancomycin, we confirmed that this was an intrinsic feature since the antibiotic resistance gene was not present. B. breve IDCC4401 showed leucine arylamidase, cystine arylamidase, α-galactosidase, β-galactosidase, and α-glucosidase activities, whereas it did not show production of harmful enzymes such as β-glucosidase and β-glucuronidase. In addition, B. breve IDCC4401 did not produce any tyramine, histamine, putrescine, cadaverine, or 2-phenethylamine, which are frequently detected BAs during fermentation. B. breve IDCC4401 produced 95.08% of L-lactate and 4.92% of D-lactate. Therefore, our findings demonstrate the safety of B. breve IDCC 4401 as a potential probiotic for use in the food industry.

α-Glucosidase Inhibitory Activity of Phenolic Compounds Isolated from the Stems of Caesalpinia decapetala var. japonica

  • Le, Thi Thanh;Ha, Manh Tuan;Hoang, Le Minh;Vu, Ngoc Khanh;Kim, Jeong Ah;Min, Byung Sun
    • Natural Product Sciences
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    • v.28 no.3
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    • pp.143-152
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    • 2022
  • In our study, sixteen known phenolic compounds, including quercetin (1), methyl gallate (2), caesalpiniaphenol C (3), 8S,8'S,7'R-(-)-lyoniresinol (4), 7,3',5'-trihydroxyflavanone (5), sappanchalcone (6), sappanone A (7), taxifolin (8), fisetin (9), fustin (10), (+)-catechin (11), brazilin (12), 3,4,5-trimethoxyphenyl β-ᴅ-glucopyranoside (13), 1-(2-methylbutyryl)phloroglucinol-glucopyranoside (14), (+)-epi-catechin (15), and astragalin (16) and one mixture of two conformers of protosappanin B (17/18) were isolated from the stems of Caesalpinia decapetala var. japonica. Their structures were elucidated based on a comparison of their physicochemical and spectral data with those of literature. To the best of our knowledge, this represents the first isolation of compounds 3, 4, 8, 9, and 10 from C. decapetala and compounds 13 and 14 from the Caesalpinia genus. All the isolated compounds were evaluated for their inhibitory effect against the α-glucosidase enzyme. Among them, two flavonols (1 and 9), one chalcone (6), and one homoisoflavanone (7) exhibited an inhibitory effect on α-glucosidase action with an IC50 range value of 5.08-15.01 μM, stronger than that of the positive control (acarbose, IC50 = 152.22 μM). Kinetic analysis revealed that compounds 1 and 9 showed non-competitive α-glucosidase inhibition, while the inhibition type was mixed for compounds 6 and 7.

Phenotypic characteristics and antimicrobial susceptibilities of motile aeromonads isolated from freshwater fish in Korea and Japan (한국과 일본의 담수어에서 분리되는 운동성 aeromonads의 표현형적 특성과 약제감수성)

  • Han, Hyun-Ja;Govindasami, Vivekanandhan;Hirono, Ikuo;Aoki, Takahashi
    • Journal of fish pathology
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    • v.20 no.3
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    • pp.249-255
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    • 2007
  • We investigated the phenotypic characteristics by using API20E, APIZYM and determined minimum inhibitory concentrations (MICs) of 7 antibiotics in motile aeromonads isolated from freshwater fishes in Korea and Japan, and 4 American Type Culture Collection (ATCC) strains. All isolates (n=7) were identified as motile Aeromonas species according to API20E test. Lysine decarboxylase activity and acid production from 4 different carbohydrates including mannitol, rhamnose, amygdalin and arabinose were observed in various strains. In enzymatic activities by APIZYM, all isolates showed negative reactions in valine and cystine arylamidases, α-chymotrypsin, α-galactosidase, β-glucuronidase, α-glucosidase, α-mannosidase and α-fucosidase. Although the intensities of each enzymatic activity were diverse in alkaline phosphatase, esterase-lipase, leucine arylamidase, β-galactosidase and N-acetyl-β-glucosaminidase, all isolates showed positive reactions. All isolates were resistant to ampicillin sodium (MIC>100㎍/ml), but sensitive to chloramphenicol (MIC≤1.6㎍/ml). However, recently isolated strains (AC9804, AC0202 and GMA0361) were commonly resistant to tetracycline (MIC=50㎍/ml). Furthermore, AC9804 was resistant to oxolinic acid (MIC=12.5㎍/ml). GMA0361 was resistant to kanamycin sulfate (MIC>100㎍/ml) and streptomycin sulfate (MIC>100 ㎍/ml).