• Title/Summary/Keyword: natural substrates

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Factors Affecting the Lactate Dehydrogenase Activity of a Spore-forming Lactic Acid Bacteria (포자형성 유산균의 lactate dehydrogenase 역가에 미치는 제요인)

  • ;Hah, Yung Chil;Hong Soon Woo;Lee, Jung Chi
    • Korean Journal of Microbiology
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    • v.15 no.3
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    • pp.103-112
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    • 1977
  • Several strains of spore-forming lacticacid bacteria were isolated from natural sources such as soils, cereals, and foods. The general morphological and physiological characteristics of the strain 6-4 were investigated nad compared with some other industrial strains. The effects of fructose-1,6-diphoshpate (FDP), adenosine triphosphate (ATP), and pH on the lactate dehydrogenase(LDH) activity of the strain were studied, and the changes in LDH activity and spore formation under various cultural conditions were researched. The results were as follows. 1. This strain was identified to Bacillus coagulans Hammer and distributed widely in natural sources. 2. The strain strongly converted various fermentation substrates in to L(+)-lacticacid in anaerobic conditioins, and many spores that were of great advantages to the industrial application were formed easily in the aerobic condition. 3. The LDH activity of this strain was activated by FDP and inhibited by ATP. The optimal pH for the enzyme activity was 6.0-6.5. 4. In the anaerobic culture condifion, the large amount of glucose added in the medium increased the LDH activity, but the cells were not committed to sporulate. 5. When none or a very small amount of glucose (less than 0.5%) was added to culture medium in the aerobic condition, the LDH activity was decreased and many spore were produced with final pH higher than 8.5. 6. The additioin of large amount of glucose (more than 2.0%) in aerobic culture increased the LDH activity and inhibited strongly the spore formation with final pH lower than 6.0.

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Characterization of growth hormone-like sequence of loach, Misgurnus mizolepis (미꾸라지 성장 호르몬 염기 서열의 특성에 대하여)

  • Kim, Jin-Kyung;Song, Young-Hwan
    • Journal of fish pathology
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    • v.7 no.2
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    • pp.95-103
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    • 1994
  • We have prepared cDNA libray of loach. M. mizolepis in order to isolate cDNA clone of growth hormone gene. Total RNA was isolated from pituitary of loach, and then mRNA was further purified from total RNA by oligo (dT)-coupled magnetic beads. The purified mRNA was used as substrates to prepare cDNA. The resulting cDNA was ligated into the EcoRV/Smal site of pBlueKS+. The ligation mixture have transformed E. coli JM109 strain with electroporator to obtain high yield of transformation efficiency. All the transformants was screened with DIG-labeled Tilapia growth hormone gene by high density colony hybridization. After isolating 10 putative colonies showing the positive signals, secondary colony hybridization and southern hybridization could confirm it as true clones. The nucleotide sequence of one candidate, pCGHI, was compared with 312 bp DNA fragment used as DNA probe and show 52% relative homology to Tilapia growth hormone gene.

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The Spawning Behavior of Korean Slender Gudgeon, Squalidus gracilis majimae, (Cypriniforms: Cyprinidae) (한국산 긴몰개 (Squalidus gracilis majimae, Cyprinidae)의 산란 행동)

  • Park, Kyung-Seo;Hong, Young-Pyo;Choi, Shin-Suk;An, Kwang-Guk
    • Korean Journal of Ecology and Environment
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    • v.38 no.2 s.112
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    • pp.207-216
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    • 2005
  • Spawning behaviors of Squalidus gracilis majimae (Cyprinidae) were observed in the laboratory whose environmental factors such as light (D/L = 16 : 8), temperature ($20\;{\sim}\;24\;^{\circ}C$), and dissolved oxygen (>8 mg $L^{-1}$) were quite regularly controlled. The behavioral patterns were categorized into three stages ofpre-spawning, spawning, and Post-spawning behaviors. In Particular, the pre-spawning stage was specified by 11 specific behavioral patterns of aggressive mating behaviors. During the spawning stage, the male and female performed four distinct spawning behaviors including sexual temptation, stimulation, egg spawning, and the separation, and randomly laid about 200 ${\sim}$ 300 eggs on the bottom substrates through the night. After finishing spawning, two adults separated toward their refuges and showed 3 types of post-spawning behaviors such as the resting, occasional eggs protecting, and the egg eating. The fish was identified as a partial-parental care species after the spawning.

Influence of Gibberellic Acid on α-D-Galactosidase Activity in the Grape Berry

  • Kang, Han-Chul;Lee, Seon-Hwa;Kim, Jong-Bum
    • Journal of Applied Biological Chemistry
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    • v.44 no.2
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    • pp.53-58
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    • 2001
  • Glycosidase activities in the grape flesh (Marguerite) were assayed, and the order of activity was marked as follows: ${\alpha}$-D-galactosidase>${\alpha}$-D-mannosidase>${\alpha}$-D-glucosidase>${\beta}$-D-galactosidase>${\beta}$-D-glucosidase. Of these glycosidases, ${\alpha}$- and ${\beta}$-D-galactosidases were prominently expressed by the treatment of gibberellic acid, resulting in 56 and 238% increase of activity, respectively. Most of ${\alpha}$-D-galactosidase was found in the flesh texture, and the activity increase by gibberellic acid occurred mostly in this tissue. The increase in ${\alpha}$-D-galactosidase activity was dependent on the concentration of gibberellic acid treated, showing a positive correlation. Gibberellic acid affected the content of total protein in the grape flesh, 49% increase by 75 ppm treatment. Above this concentration, higher gibberellic acid level did not influence the protein expression. Specific activity of the ${\alpha}$-D-galactosidase still increased, showing 24% increase in activity. Grape flesh subjected by gibberellic acid (100 ppm) resulted in the increased activity against a natural substrate, stachyose, showing 55% increase in activity from the grapes treated with 100 ppm of gibberellic acid. Other natural substrates, such as melibiose and raffinose, were also considerably hydrolyzed, and the extent was similar to that of stachyose hydrolysis. During postharvest storage, ${\alpha}$-D-galactosidase activity in the grape flesh increased by 51% after 20 days and then declined slowly.

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Cloning and Expression of a Thermostable ${\alpha}$-Galactosidase from the Thermophilic Fungus Talaromyces emersonii in the Methylotrophic Yeast Pichia pastoris

  • Simila, Janika;Gernig, Anita;Murray, Patrick;Fernandes, Sara;Tuohy, Maria G.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1653-1663
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    • 2010
  • The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

The effect of plant extracts on the activity and the expression of MMPs (matrix metalloprotease) induced by UVA

  • Lee, Dong-hwan;Lee, Bum-chun;Yoon, Eun-jeong;Lee, Kyung-eun;Park, Sung-min;Pyo, Hyeong-bae;Choe, Tae-boo
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.32-43
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    • 2003
  • UV irradiation on a skin brings about the qualitative and quantitative alterations of the extracellular matrix. Repeated-UV irradiation suppressed the synthesis of collagen and activated the expression of the matrix metalloprotease (MMP). In this paper, on the purpose of development of novel anti-aging agents from natural sources, effects of several natural products on in vitro MMP-1 activity and UVA induced MMP-1 synthesis in human dermal fibroblast (HDF) culture were studied. We measured MMP-1 activities by fluorescence assay using gelatin as substrates. As a result, the extract of Dicentra spectabilis, and flower buds of Tussilago farfara showed strong inhibitory effect. Among them, the extract of flower buds of Tussilago fartara and Dicentra spectabilis inhibited MMP-1 activity by 92% and 87% at 0.05% (w/v). And UVA induced MMP-1 expression were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in HDF culture. The extract of flower buds of Tussilago farfara and Dicentra spectabilis suppressed the UVA induced expression of MMP-1 by similar level of Vitamin C 200$\mu$M at 0.1% (w/v). These results suggest that the extract of Dicentra spectabilis, and flower buds of Tussilago farfara effectively prevent skin from the UV-induced photoaging. So the extracts are thought to have potential as effective raw materials for anti-aging cosmetics.

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Metagenomic SMRT Sequencing-Based Exploration of Novel Lignocellulose-Degrading Capability in Wood Detritus from Torreya nucifera in Bija Forest on Jeju Island

  • Oh, Han Na;Lee, Tae Kwon;Park, Jae Wan;No, Jee Hyun;Kim, Dockyu;Sul, Woo Jun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1670-1680
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    • 2017
  • Lignocellulose, composed mostly of cellulose, hemicellulose, and lignin generated through secondary growth of woody plant, is considered as promising resources for biofuel. In order to use lignocellulose as a biofuel, biodegradation besides high-cost chemical treatments were applied, but knowledge on the decomposition of lignocellulose occurring in a natural environment is insufficient. We analyzed the 16S rRNA gene and metagenome to understand how the lignocellulose is decomposed naturally in decayed Torreya nucifera (L) of Bija forest (Bijarim) in Gotjawal, an ecologically distinct environment. A total of 464,360 reads were obtained from 16S rRNA gene sequencing, representing diverse phyla; Proteobacteria (51%), Bacteroidetes (11%) and Actinobacteria (10%). The metagenome analysis using single molecules real-time sequencing revealed that the assembled contigs determined originated from Proteobacteria (58%) and Actinobacteria (10.3%). Carbohydrate Active enZYmes (CAZy)- and Protein families (Pfam)-based analysis showed that Proteobacteria was involved in degrading whole lignocellulose, and Actinobacteria played a role only in a part of hemicellulose degradation. Combining these results, it suggested that Proteobacteria and Actinobacteria had selective biodegradation potential for different lignocellulose substrates. Thus, it is considered that understanding of the systemic microbial degradation pathways may be a useful strategy for recycle of lignocellulosic biomass, and the microbial enzymes in Bija forest can be useful natural resources in industrial processes.

Anti-inflammatory effect of Distylium racemosum leaf biorenovate extract in LPS-stimulated RAW 264.7 macrophages cells (LPS로 유도된 RAW 264.7 세포에 대한 조록나무 잎 Biorenovation 추출물의 항염증 활성)

  • Hong, Hyehyun;Lee, Kyung-Mi;Park, Taejin;Chi, Won-Jae;Kim, Seung-Young
    • Journal of Applied Biological Chemistry
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    • v.64 no.4
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    • pp.375-382
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    • 2021
  • Biorenovation is a microbial enzyme-based structural modification of component compounds in natural products and synthetic compounds including plant extracts with the potential benefits of improved biological activities compared with its reaction substrates. In this study, we investigated the anti-inflammatory activity of Distylium racemosum leaf extract and D. racemosum leaf biorenovation extract (DLB). As a result, DLB inhibited nitric oxide, prostaglandin E2, and inflammatory cytokines including tumor necrosis factor-α, interleukin-6, interleukin-1β at non-toxic concentrations. In addition, DLB significantly inhibited inducible nitric oxide synthase and cyclooxygenase-2 on LPS-treated RAW 264.7 macrophages. Based on these results, we suggest that the DLB could be used as a potent anti-inflammatory agents. It also suggests that the application of biological evolution has potential usefulness to increase the practical value of natural products.

Effect of Shell-type, Light and Temperature on the Shell Infiltration of Free-living Conchocelis of Three Pyropia Species (김(Pyropia spp.) 3종 유리사상체의 패각 잠입에 대한 패각 종류, 광과 온도의 영향)

  • Heo, Jin Suk;Park, Eun Jung;Hwang, Mi Sook;Choi, Han Gil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.1
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    • pp.23-30
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    • 2021
  • To examine the optimal temperature, light intensity, and shell-type for shell-living conchocelis production, we tested the shell infiltration of free-living conchocelis fragments under various environmental conditions. Under a combination of various temperatures (10, 15, 20, 25 and 30℃) and light intensities (1, 5, 10, 20, 40, and 80 μmol m-2 s-1), the optimal infiltration conditions of the evaluated three Pyropia species were 20-25℃ and 5-80 μmol m-2 s-1 for P. yezoensis, 20-30℃ and 20-80 μmol m-2 s-1 for P. seriata, and 20-25℃ and 20-80 μmol m-2 s-1 for P. dentata. The infiltration efficiency of free-living conchocelis for different shell types was greater in Korean and Chinese oyster Crassostrea gigas shells than that in scallop Argopecten irradians and clam Meretrix lusoria shells. These results suggest that oyster shells are suitable substrates for shell-living conchocelis production. In conclusion, the present results for optimal infiltration conditions for free-living conchocelis of the three examined Pyropia species will contribute significantly to the production of stable shell-living conchocelis.

Characterization and Production of Low Molecular Weight of Biopolymer by Weisella sp. strain YSK01 Isolated from Traditional Fermented Foods (전통 발효식품으로부터 분리된 Weisella sp. strain YSK01에 의한 저분자 Biopolymer 발효생산 공정 및 생성물의 특성)

  • Cho, Hyun Ah;Kim, Nam Chul;Yoo, Sun Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.5
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    • pp.632-643
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    • 2022
  • Although probiotics have been shown to improve health when consumed, recent studies have reported that they can cause unwanted side effects due to bacterial-human interactions. Therefore, the importance of prebiotics that can form beneficial microbiome in the gut has been emphasized. This study isolated and identified bacteria capable of producing biopoymer as a candidate prebiotic from traditional fermented foods. The isolated and identified strain was named WCYSK01 (Wissella sp. strain YSK01). The composition of the medium for culturing this strain was prepared by dissolving 3 g K2HPO4, 0.2 g MgSO4, 0.05 g CaCl2, 0.1 g NaCl in 1 L of distilled water. The LMBP(low molecular weight biopoymers) produced when fermentation was performed with sucrose and maltose as substrates were mainly consisted of DP3 (degree of polymer; isomaltotriose), DP4 (isomaltotetraose), DP5 (isomaltopentaose), and DP6 (isomaltoheptaose). The optimization of LMBP (low molecular weight of biopolymer) production was performed using the response surface methodology. The fermentation process temperature range of 18 to 32℃, the fermentation medium pH in the range of 5.1 to 7.9. The yield of LMBP production by the strain was found to be significantly affected by q fermentation temperature and pH. The optimal fermentation conditions were found at the normal point, and the production yield was more than 75% at pH 7.5 and temperature of 23℃.