• Title/Summary/Keyword: Microbial Enzymatic Activity

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Optimal Production of N-acetyl-$\beta$-D-glucosamine Using Chitinolytic Enzyme (Chitinolytic Enzyme을 이용한 N-acetyl-$\beta$-D-glucosamine의 최적생산)

  • 이천우;이은영장상목김광
    • KSBB Journal
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    • v.11 no.6
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    • pp.696-703
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    • 1996
  • The bacterium Serratia marcescens QM Bl466 produces selectively large amount of chitinolytic enzymes(about 1mg/L medium). Enzymatic hydrolysis of chitin to N-acelyl-${\beta}$-D-glucosamine(NAG) is performed by a system consisting of two hydrolases : chitinase and chilobiase. Objectives of this study included optimization of a microbial host by using chitin particles for chitinase/chitobiase production and secretion and also development of batch fermentation system for high cell density cultivalion of S. marcescens QM B1466. Also, the influence of chitin source and carboxymethyl(CM) chitin on chitinase/chitobiase production and NAG production was investigated. When carboxymethyl chitin was substituted for colloidal and practical grade chitin, the chitinase activity was increased about 7∼10U/mL. In this case, the ratio of chitinase/chitobiase was 30.03U/3.44U(9:1). The highest amounts of NAG(3.0g/L) was obtained.

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Fermentation of red ginseng extract by the probiotic Lactobacillus plantarum KCCM 11613P: ginsenoside conversion and antioxidant effects

  • Jung, Jieun;Jang, Hye Ji;Eom, Su Jin;Choi, Nam Soon;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.20-26
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    • 2019
  • Background: Ginsenosides, which are bioactive components in ginseng, can be converted to smaller compounds for improvement of their pharmacological activities. The conversion methods include heating; acid, alkali, and enzymatic treatment; and microbial conversion. The aim of this study was to determine the bioconversion of ginsenosides in fermented red ginseng extract (FRGE). Methods: Red ginseng extract (RGE) was fermented using Lactobacillus plantarum KCCM 11613P. This study investigated the ginsenosides and their antioxidant capacity in FRGE using diverse methods. Results: Properties of RGE were changed upon fermentation. Fermentation reduced the pH value, but increased the titratable acidity and viable cell counts of lactic acid bacteria. L. plantarum KCCM 11613P converted ginsenosides $Rb_2$ and $Rb_3$ to ginsenoside Rd in RGE. Fermentation also enhanced the antioxidant effects of RGE. FRGE reduced 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power; however, it improved the inhibition of ${\beta}$-carotene and linoleic acid oxidation and the lipid peroxidation. This suggested that the fermentation of RGE is effective for producing ginsenoside Rd as precursor of ginsenoside compound K and inhibition of lipid oxidation. Conclusion: This study showed that RGE fermented by L. plantarum KCCM 11613P may contribute to the development of functional food materials.

Psychobiotic Effects of Multi-Strain Probiotics Originated from Thai Fermented Foods in a Rat Model

  • Luang-In, Vijitra;Katisart, Teeraporn;Konsue, Ampa;Nudmamud-Thanoi, Sutisa;Narbad, Arjan;Saengha, Worachot;Wangkahart, Eakapol;Pumriw, Supaporn;Samappito, Wannee;Ma, Nyuk Ling
    • Food Science of Animal Resources
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    • v.40 no.6
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    • pp.1014-1032
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    • 2020
  • This work aimed to investigate the psychobiotic effects of six bacterial strains on the mind and behavior of male Wistar rats. The probiotic (PRO) group (n=7) were rats pre-treated with antibiotics for 7 days followed by 14-day probiotic administration, antibiotics (ANT) group (n=7) were rats treated with antibiotics for 21 days without probiotics. The control (CON) group (n=7) were rats that received sham treatment for 21 days. The six bacterial strains with probiotic properties were mostly isolated from Thai fermented foods; Pedicoccus pentosaceus WS11, Lactobacillus plantarum SK321, L. fermentum SK324, L. brevis TRBC 3003, Bifidobacterium adolescentis TBRC 7154 and Lactococcus lactis subsp. lactis TBRC 375. The probiotics were freeze-dried into powder (6×109 CFU/5 g) and administered to the PRO group via oral gavage. Behavioral tests were performed. The PRO group displayed significantly reduced anxiety level and increased locomotor function using a marble burying test and open field test, respectively and significantly improved short-term memory performance using a novel object recognition test. Antibiotics significantly reduced microbial counts in rat feces in the ANT group by 100 fold compared to the PRO group. Probiotics significantly enhanced antioxidant enzymatic and non-enzymatic defenses in rat brains as assessed using catalase activity and ferric reducing antioxidant power assay, respectively. Probiotics also showed neuroprotective effects with less pyknotic cells and lower frequency of vacuolization in cerebral cortex. This multi-strain probiotic formulation from Thai fermented foods may offer a potential to develop psychobiotic-rich functional foods to modulate human mind and behaviors.

Target Identification for Metabolic Engineering: Incorporation of Metabolome and Transcriptome Strategies to Better Understand Metabolic Fluxes

  • Lindley, Nic
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.60-61
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    • 2004
  • Metabolic engineering is now a well established discipline, used extensively to determine and execute rational strategies of strain development to improve the performance of micro-organisms employed in industrial fermentations. The basic principle of this approach is that performance of the microbial catalyst should be adequately characterised metabolically so as to clearlyidentify the metabolic network constraints, thereby identifying the most probable targets for genetic engineering and the extent to which improvements can be realistically achieved. In order to harness correctly this potential, it is clear that the physiological analysis of each strain studied needs to be undertaken under conditions as close as possible to the physico-chemical environment in which the strain evolves within the full-scale process. Furthermore, this analysis needs to be undertaken throughoutthe entire fermentation so as to take into account the changing environment in an essentially dynamic situation in which metabolic stress is accentuated by the microbial activity itself, leading to increasingly important stress response at a metabolic level. All too often these industrial fermentation constraints are overlooked, leading to identification of targets whose validity within the industrial context is at best limited. Thus the conceptual error is linked to experimental design rather than inadequate methodology. New tools are becoming available which open up new possibilities in metabolic engineering and the characterisation of complex metabolic networks. Traditionally metabolic analysis was targeted towards pre-identified genes and their corresponding enzymatic activities within pre-selected metabolic pathways. Those pathways not included at the onset were intrinsically removed from the network giving a fundamentally localised vision of pathway functionality. New tools from genome research extend this reductive approach so as to include the global characteristics of a given biological model which can now be seen as an integrated functional unit rather than a specific sub-group of biochemical reactions, thereby facilitating the resolution of complexnetworks whose exact composition cannot be estimated at the onset. This global overview of whole cell physiology enables new targets to be identified which would classically not have been suspected previously. Of course, as with all powerful analytical tools, post-genomic technology must be used carefully so as to avoid expensive errors. This is not always the case and the data obtained need to be examined carefully to avoid embarking on the study of artefacts due to poor understanding of cell biology. These basic developments and the underlying concepts will be illustrated with examples from the author's laboratory concerning the industrial production of commodity chemicals using a number of industrially important bacteria. The different levels of possibleinvestigation and the extent to which the data can be extrapolated will be highlighted together with the extent to which realistic yield targets can be attained. Genetic engineering strategies and the performance of the resulting strains will be examined within the context of the prevailing experimental conditions encountered in the industrial fermentor. Examples used will include the production of amino acids, vitamins and polysaccharides. In each case metabolic constraints can be identified and the extent to which performance can be enhanced predicted

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Bacterial Numbers and Exoenzymatic Activities in Pore Water of Artificial Floating Island Installed in Lake Paldang (팔당호 인공식물섬 공극수에서 미생물 개체수와 체외효소활성도)

  • Kim, Yong-Jeon;Choi, Seung-Ik;Ahn, Tae-Seok
    • Korean Journal of Ecology and Environment
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    • v.41 no.1
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    • pp.19-25
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    • 2008
  • To evaluate the functions of vegetation mat of artificial floating island (AFI) installed in Lake Paldang, nutrients, such as total phosphorus (TP), dissolved inorganic phosphorus (DIP), total nitrogen (TN) and nitrate $(NO_3)$ and microbial factors such as total bacterial numbers, active bacterial numbers and exoenzymatic activities of $\beta$-glucosidase and phosphatase in pore water of medium and bulk lake water were analyzed. The concentration of TN and $NO_3$ in pore water ranged from 4.4 to 7.5mg $L^{-1}$ from 1.2 to 3.8mg $L^{-1}$ respectively, which were ca. 2 times higher than those of lake water. The ranges of TP and DIP of were $1.4\sim4.1mg\;L^{-1}$ and $0.003\sim0.137mg\;L^{-1}$ in pore water of media which were $4\sim25$ and 5 times higher than those of lake water, respectively. The numbers of total bacteria and active bacteria in pore waterwere about 10 times higher than those of laker water. Also, both phosphatase and $\beta$-glucosidase activities of pore water were on an average 10 times higher than those of lake water. These results suggest that the bacteria were playing important role for nutrients concentrating and cycling in media of artificial floating island. And the medium of artificial floating island contained newly created microbial ecosystem, which is responsible for sustaining the growth of macrophytes and the creation of new aquatic ecosystem.

Enzymological Properties of the Alkaline AL-Protease from Arthrobacter luteus and Detection of Its Active Amino Acid Residue (Arthrobacter luteus로부터 유래한 염기성 AL-Protease의 효소학적 성질 및 활성 아미노산 잔기의 검색)

  • Oh, Hong-Rock;Aizono, Yasuo;Funatsu, Masaru
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.13 no.2
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    • pp.193-204
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    • 1984
  • The enzymatic properties of the alkaline AL-protease, which had been prepared from the crude zymolyase of Arthrobzoter luteus, was investigated together with its active amino acid residue. Complete inactivaton of the proteolytic activity of AL-protease by either DFP or PMSF was simultaneously accompanied by the loss of its lytic effect on the lysis of yeast cell wall. In the reaction, AL-protease showed the pattern of inactivation to decrease very slowly, as compared to that of chymotrypsin, and that enzyme and DFP were found to react with a molar ratio of 1 : 1. The preparation of AL-protease exhibited no hydrolytic activity in any substrates of polysaccharases, playing a significant role in the lysis of yeast cell wall. The optimum pH and temperature of AL-protease was pH 10.5 and $65^{\circ}C$, respectively. It also showed stability in the pH range from 5 to 11 and at the temperature below $65^{\circ}C$. Through the identification of the amino acid residue in the active site of the $^{32}P$-diisopropylph-osphorylated(DIP) AL-protease modified specifically with $^{32}P$-labeled DFP, AL-protease was found to be a DFP-sensitive which has a mole of active serine residue involved in its proteolytic activity per mole of the enzyme.

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A review of factors that regulate extracellular enzyme activity in wetland soils (습지 토양 내 체외효소 활성도를 조절하는 인자에 대한 고찰)

  • Kim, Haryun
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.97-107
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    • 2015
  • Wetlands constitute a transitional zone between terrestrial and aquatic ecosystems and have unique characteristics such as frequent inundation, inflow of nutrients from terrestrial ecosystems, presence of plants adapted to grow in water, and soil that is occasionally oxygen deficient due to saturation. These characteristics and the presence of vegetation determine physical and chemical properties that affect decomposition rates of organic matter (OM). Decomposition of OM is associated with activities of various extracellular enzymes (EE) produced by bacteria and fungi. Extracellular enzymes convert macromolecules to simple compounds such as labile organic carbon (C), nitrogen (N), phosphorus (P), and sulfur (S) that can be easily taken up by microbes and plants. Therefore, the enzymatic approach is helpful to understand the decomposition rates of OM and nutrient cycling in wetland soils. This paper reviews the physical and biogeochemical factors that regulate extracellular enzyme activities (EEa) in wetland soils, including those of ${\beta}$-glucosidase, ${\beta}$-N-acetylglucosaminidase, phosphatase, arylsulfatase, and phenol oxidase that decompose organic matter and release C, N, P, and S nutrients for microbial and plant growths. Effects of pH, water table, and particle size of OM on EEa were not significantly different among sites, whereas the influence of temperature on EEa varied depending on microbial acclimation to extreme temperatures. Addition of C, N, or P affected EEa differently depending on the nutrient state, C:N ratio, limiting factors, and types of enzymes of wetland soils. Substrate quality influenced EEa more significantly than did other factors. Also, drainage of wetland and increased temperature due to global climate change can stimulate phenol oxidase activity, and anthropogenic N deposition can enhance the hydrolytic EEa; these effects increase OM decomposition rates and emissions of $CO_2$ and $CH_4$ from wetland systems. The researches on the relationship between microbial structures and EE functions, and environmental factors controlling EEa can be helpful to manipulate wetland ecosystems for treating pollutants and to monitor wetland ecosystem services.

Fermentation Characteristics of Low Salted Kochujang Prepared with Sub-materials (부원료를 첨가한 저식염 고추장의 발효특성 변화)

  • Kim, Dong-Han;Yang, Sung-Eun
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.97-104
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    • 2004
  • Effects of sub-materials such as, ethanol, mustard, and chitosan, on enzymatic, microbial and physicochemical characteristics of kochujang were investigated during fermentation. Activity of ${\alpha}$-amylase was low in chitosan-added kochujang, whereas those of ${\beta}$-amylase and pretense did not show any remarkable difference. Viable cells of yeast and bacteria decreased in sub-material-added kochujang during fermentation, with yeast counts decreasing more rapidly in ethanol- and mustard-added kochujang than that with chitosan. Consistency of kochujang decreased during fermentation, with the highest consistency observed in ethanol-added kochujang. Oxidation-reduction potential was low in chitosan-added kochujang. Water activity of all kochujang groups decreased during fermentation with the lowest in ethanol-added kochujang. Hunter L-, a-, and b-values of chitosan-added kochujang were higher than other groups, whereas increase in total color difference of low-salt without sub-material group was lowest pH of kochujang was the highest in ethanol-added kochujang, whereas titratable acidity increased remarkably in chitosan-added group. Total sugar and reducing sugar contents of kochujang were high in ethanol-and mustard-added groups, whereas ethanol production decreased remarkably in mustard-added kochujang. Amino nitrogen content was highest in low-salt without sub-material kochujang during late aging period. Ammonia nitrogen content was lower in mustard-added kochujang. After 12 weeks of fermentation, ethanol-added kochujang was more acceptable than mustard-and chitosan-added groups in taste, color, and overall acceptabilities.

Fermentation Characteristics of Kochujang Prepared with Various Salts (소금의 종류를 달리한 고추장의 발효 특성)

  • Kim, Dong-Han;Yang, Sung-Eun;Rhim, Jong-Whan
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.671-679
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    • 2003
  • Effects of sun-dried, refined, seaweed, and bamboo salts on the quality of kochujang were studied by measuring enzymatic, microbial, and physicochemical characteristics of kochujang during 12 weeks of fermentation. Yeast count was low in the bamboo-salt kochujang, whereas that of aerobic bacteria was low in the seaweed-salt kochujang. Acid protease activity was high in the bamboo-salt kochujang, whereas amylase activity did not show any remarkable difference. Color change was lowest in the bamboo-salt kochujang. Water activities of all kochujangs decreased during fermentation with the lowest shown in the refined-salt kochujang. Consistency of seaweed-salt kochujang was the highest. Total sugar content was higher, whereas ORP was lower in the seaweed- and bamboo-salt kochujangs. Titratable acidity changed slightly in the bamboo-salt kochujang. Reducing sugar content was the lowest, whereas ethanol content was the highest in the refined-salt kochujang. Amino and ammonia nitrogen contents of kochujangs increased up to the middle of fermentation period then decreased with lower changes observed in the sun-dried and refined-salt kochujangs. Results of sensory evaluation indicated that the bamboo-salt kochujang was more acceptable than the sun-dried salt kochujang in taste, color, and overall acceptabilities.

The In vitro Effects of Nano-encapsulated Conjugated Linoleic Acid on Stability of Conjugated Linoleic Acid and Fermentation Profiles in the Rumen

  • Heo, Wan;Kim, Eun Tae;Cho, Sung Do;Kim, Jun Ho;Kwon, Seong Min;Jeong, Ha Yeon;Ki, Kwang Seok;Yoon, Ho Baek;Ahn, Young Dae;Lee, Sung Sill;Kim, Young Jun
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.365-371
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    • 2016
  • This study was aimed to evaluate the stability of conjugated linoleic acids (CLAs) by nano-encapsulation against in vitro ruminal biohydrogenation by microbial enzymatic conversion. CLAs (free fatty acid form of CLA [CLA-FFA], nano-encapsulated CLA-FFA, triglyceride form of CLA [CLA-TG], and nano-encapsulated CLA-TG) were used in the in vitro fermentation experiments. When Butyrivibrio fibrisolvens (B. fibrisolvens) was incubated with CLA-FFAs, the concentrations of cis-9, trans-11 CLA and vaccenic acid (VA) slightly was decreased and increased by nano-encapsulation, respectively. When B. fibrisolvens was incubated with CLA-TG, the concentrations of cis-9, trans-11 CLA and VA decreased, but these were increased when B. fibrisolvens was incubated with nano-encapsulated CLA-TG. The nano-encapsulation was more effective against the in vitro biohydrogenation activity of B.fibrisolvens incubated with CLA-FFA than with CLA-TG. In the in vitro ruminal incubation test, the total gas production and concentration of total volatile fatty acids incubated with nano-encapsulated CLA-FFA and CLA-TG were increased significantly after 24 h incubation (p<0.05). Nano-encapsulated CLA-FFA might, thus, improve the ruminal fermentation characteristics without adverse effects on the incubation process. In addition, nano-encapsulated CLA-FFA increased the population of Fibrobacter succinogenes and decreased the population of B. fibrisolvens population. These results indicate that nano-encapsulation could be applied to enhance CLA levels in ruminants by increasing the stability of CLA without causing adverse effects on ruminal fermentation.