• Title/Summary/Keyword: microbial enzyme

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The Effect of Enzyme/Microbial Additive on Anaerobic Digestion of Primary Sludge

  • Kim, Hyung-Jin;Song, Chang-Soo;Kim, Dong-Wook;Pagilla, Kishna-R.
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.10 no.S_1
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    • pp.35-40
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    • 2001
  • Effect of the addition of an enzyme/microbial additive(EMA) to enhance anaerobic digestion of the primary sludge was investigated. Two laboratory scale anaerobic digester were operated with primary sludge taken from a municipal wastewater treatment plant. The digester receiving EMA with the sludge feed performed better than the control digester, when both were operated at 10-days and 15-days Solid Retention Time(SRT). Addition of EMA to the experimental digester provided 7%(10-days SRT) and 16%(15-days SRT) higher gas production compared to the control digester when both were fed with the same amount of volatile solids. The reduction in volatile solids was 24% better in the experimental digester compared to the control ar 10-days SRT, and the improvement 10% at 15-day SRT. Improvement in COD reduction, and fecal coliform density reduction were also seen in the experimental digester due to EMA addition compared to the control both ar 10-days SRT and 15-day SRT operation. Preliminary cost benefit analysis for a wastewater treatment plant showed that approximately $115/day in gas production improvements can be realized upon addition of EMA to primary sludge anaerobic digesters operating at 10-day SRT. The value of increased gas production was $172/day if the same digesters are operated with EMA addition at 15-day SRT.

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Characterization of Calcium-Activated Bifunctional Peptidase of the Psychrotrophic Bacillus cereus

  • Kim Jong-Il;Lee Sun-Min;Jung Hyun-Joo
    • Journal of Microbiology
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    • v.43 no.3
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    • pp.237-243
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    • 2005
  • The protease purified from Bacillus cereus JH108 has the function of leucine specific endopeptidase. When measured by hydrolysis of synthetic substrate (N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide), the enzyme activity exhibited optimal activity at pH 9.0, $60^{\circ}C$. The endopeptidase activity was stimulated by $Ca^{++},\;Co^{++},\;Mn^{++},\;Mg^{++},\;and\;Ni^{++}$, and was inhibited by metal chelating agents such as EDTA, 1,10-phenanthroline, and EGTA. Addition of serine protease inhibitor, PMSF, resulted in the elimination of the activity. The endopeptidase activity was fully recovered from the inhibition of EDTA by the addition of 1 mM $Ca^{++}$, and was partially restored by $Co^{++}\;and\;Mn^{++}$, indicating that the enzyme was stabilized and activated by divalent cations and has a serine residue at the active site. Addition of $Ca^{++}$ increased the pH and heat stability of endopeptidase activity. These results show that endopeptidase requires calcium ions for activity and/or stability. A Lineweaver-Burk plot analysis indicated that the $K_m$ value of endopeptidase is 0.315 mM and $V_{max}$ is 0.222 ) is $0.222\;{\mu}mol$ of N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide per min. Bestatin was shown to act as a competitive inhibitor to the endopeptidase activity.

Differences in Biogeochemical Properties and Microbial Activities in Stream Segments with Changes in Land-use Type

  • Kim, Jinhyun;Jang, Inyoung;Lee, Hyunjin;Kang, Hojeong
    • Ecology and Resilient Infrastructure
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    • v.2 no.3
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    • pp.247-254
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    • 2015
  • Changes in land-use type can affect soil and water properties in stream ecosystems. This study examined the effects of different land-use types on biogeochemical properties and microbial activities of a stream. We collected water and sediment samples in a stream at three different sites surrounded by varying land-use types; a forest, a radish field and a rice paddy. Nitrogen contents, such as nitrate, nitrite and total nitrogen in the stream water body, showed significant differences among the sampling sites. The highest nitrogen values were recorded at the site surrounded by cropland, as fertilizer runoff impacted the stream. Soil organic matter content in the sediment showed significant differences among sites, with the highest content exhibited at the forest mouth site. These differences might be due to the organic matter in surrounding terrestrial ecosystems. Microbial activities determined by extracellular enzyme activities showed similar values throughout all sites in the water body; however, the activities in the sediments exhibited the highest values near the forest site and mirrored the soil organic matter content values. From these results, we conclude that different land-use types are important factors affecting water and sediment properties in stream ecosystems.

Extraction of β-glucosidase from Bagasse Fermented by Mixed Culture under Solid State Fermentation

  • Shata, Hoda Mohamed Abdel Halim;Farid, Mohamed Abdel Fattah
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.197-203
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    • 2014
  • Various parameters such as solvent selection, concentration, solid/liquid ratio, soaking time, temperature, stationary, shaking conditions, and repeated extractions were investigated in order to determine the optimum extraction conditions of ${\beta}$-glucosidase from bagasse fermented by mixed culture of Aspergillus niger NRC 7A and Aspergillus oryzae NRRL 447. Among various solvents tested, non ionic detergents gave the best results than the inorganic or organic salt solutions and distilled water. The optimum conditions for extraction of ${\beta}$-glucosidase were 30 min soaking time at $40^{\circ}C$ under shaking condition at 150 rpm, with solid/liquid ratio 1:15 (w/v), which yielded $2882.74{\pm}95.52U/g$ fermented culture (g fc) of enzyme activity. With repeated washes under the above optimum conditions, the results showed that enzyme extracted in the $1^{st}$ and $2^{nd}$ washes represents about 90% of the total activity.

Potential Application of the Recombinant Escherichia coli-Synthesized Heme as a Bioavailable Iron Source

  • Kwon, Oh-Hee;Kim, Su-Sie;Hahm, Dae-Hyun;Lee, Sang-Yup;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • v.19 no.6
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    • pp.604-609
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    • 2009
  • To investigate the potential use of microbial heme as an iron source, recombinant Escherichia coli coexpressing ALA synthase (HemA) as well as the NADP-dependent malic enzyme (MaeB) and dicarboxylic acid transporter (DctA) were cultured. The typical red pigment extracted from the recombinant E. coli after 38 h showed highest absorbance at 407 nm, and the amount of iron in 38.4 mg of microbial heme extract derived from 6-1 fermentation broth was 4.1 mg. To determine the commercial potential of the recombinant E.coli-synthesized iron-associated heme as an iron source, mice were fed the iron-free provender with the microbial heme extract. The average body weight reduction of mice fed non-iron provender was 2.3%, whereas no detectable weight loss was evident in mice fed microbial heme addition after 15 days. The heme content of the blood from microbial heme fed mice was 4.2 mg/ml whereas that of controls was 2.4 mg/ml, which implies that the microbial heme could be available for use as an animal iron source.

Effect of Fire on Microbial Community Structure and Enzyme Activities in Forest Soil (산불이 토양 미생물 군집과 효소 활성 변화에 미치는 영향)

  • Oh, Ju-Hwan;Lee, Seul-Bi;Park, Sung-Eun;Lee, Yong-Bok;Kim, Pil-Joo
    • Korean Journal of Environmental Agriculture
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    • v.27 no.2
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    • pp.133-138
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    • 2008
  • Fire can affect microbial community structure of soil through altered environmental conditions, nutrient availability, and biotic source for microbial re-colonization. We examined the influence of fire on chemical properties and soil enzyme activities of soil for 10 months. We also characterized the soil microbial community structure through ester-linked fatty acid analysis(EL-FAME). For this study, we established five burned plots(1*1 m) and 5 unburned plots outside the margin of fire. Soil was sampled three soil cores in a each plots and composited for analysis at 1, 3, 5, 8, and 10 month after fire. The fire caused an increase in soil pH, exchangeable Ca, and Mg, organic matter, available $P_2O_5$ compared to unburned sites. The content of $NH_4-N$ in burned site was significantly higher than that of unburned site and this effect continued for 8 months after fire. There was no difference of $NO_3-N$ content in soil between burned and unburned site. Fire caused no change in acid phosphatase and arylsulfatase activities but $\beta$-glucosidase and alkaline phosphatase activities in burned site were increased compared to unburned site. Microbial biomass as estimated by total concentration of EL-FAMEs in burned sites was significantly higher than that of unburned sites at one month after fire. Burned site decreased the EL-FAMEs indicative of gram-positive bacteria and tended to increase the fatty acid associated with gram-negative bacteria at one and three months after fire. The sum of EL-FAME compound $18:2{\omega}6,9c$ and $18:1{\omega}9c$ as served fungal biomarkers was decreased in burned site compared to unburned site.

Effects of Grubbing by Wild Boars on the Biological Activities of Forest Floor (멧돼지(Sus scrofa coreanus Heude)의 임상 교란이 토양의 생물학적 활성에 미치는 영향)

  • Cha, Sangsub;Lee, Sang-Hoon;Chae, Hee-Myung;Shim, Jae Kuk
    • Korean Journal of Environment and Ecology
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    • v.26 no.6
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    • pp.902-910
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    • 2012
  • This study has been carried out to determine the effects of disturbances from wild boar grubbing on the functions of ecosystem. The experiments was performed in Mt. Jumbong of Long-term Ecological Research Sites of the Ministry of Environment. We measured soil physical properties, soil respiration($CO_2$), microbial biomass C, and soil enzyme activities from both disturbed and control plots. The disturbance sites were divided into two parts, mounds and pits. Soil organic matter contents were highest value at the control plots and lowest at the pit plots, respectively at 20.22% and 15.52%. The soil bulk densities were highest at the pit plots. Soil microbial biomass C and $CO_2$ evolution were significantly higher at the control plots compared to the disturbed plots. The results were positively correlated with soil organic matter contents. The cellulase activity and invertase activity in the soil showed similar pattern as the microbial biomass C and $CO_2$ evolution results. The cellulase activity and invertase activity in the soil were positively correlated with soil microbial biomass C. Soil organic matter contents seemed to affect the soil enzyme activities. The nitrate reductase activities were highest at the pit plots, which showed positive correlation with soil bulk density. The study results showed that the grubbing disturbances by wild boars induced the changes in soil properties, which affected soil microbial activities.

Screening and Characterization of Lactate Dehydrogenase-producing Microorganism

  • Sung, Ha Guyn;Lee, Jae Heung;Shin, Hyung Tai
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.10
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    • pp.1411-1416
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    • 2004
  • The objective of this work was to isolate a microorganism, able to produce high lactate dehydrogenase (LDH) activity, for use as a microbial feed additive. The LDH is an important enzyme for lactate conversion in the rumen, thereby possibly overcoming lactic acidosis owing to sudden increases of cereal in the diets of ruminants. In the present study, various bacterial strains were screened from a variety of environments. Among the isolated microorganisms, strain FFy 111-1 isolated from a Korean traditional fermented vegetable food called Kimchi showed the highest enzyme activity, along with retaining strong enzyme activity even in rumen fluid in vitro. Based on morphological and biochemical characteristics as well as compositions of cellular fatty acids plus API analyses, this strain was identified as Lactobacillus sp. The optimum temperature and pH for growth were found to be 30$^{\circ}C$ and pH 6.5, respectively. A maximum cell growth of 2.2 at $A_{650}$ together with LDH activity of 2.08 U per mL was achieved after 24 h of incubation. Initial characterization of FFy 111-1 suggested that it could be a potential candidate for use as a direct-fed microbial in the ruminant animals.

Purification and Characterization of Lipase from Trichosporon sp. Y-11and Its Use in Ester Synthesis of Unsaturated Fatty Acids and Alcohols

  • Song, Xin;Qu, Yinbo;Shin, Dong-Hoon;Kim, Eun-Ki
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.951-956
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    • 2001
  • A 28-kDa extracellular lipase (pI 8.7) was purified to homogeneity from the culture supernatant of Trichosporon sp. Y- 11 by mmonium sulfate precipitation, DEAE-Sephadex A-50, Bio-Gel P-30, CM- Sephadex C-50, and Bio-Gel P- 10 chromatographies. The purified enzyme exhibited a specific activity of $2,741{\;}{\mu}mol/min/mg$ based on the hydrolysis of triolein, and the optimal hydrolysis activity was dentified at pH 8.0 and $40^{\circ}C$. The enzyme activity was inhibited by $Ag^+$ and enhanced by $Fe^{2+}$, $Fe^{3+}$, $Mg^{2+}$, $Mn^{2+}$, and $Li^{+}$. The enzyme activity exhibited for the hydrolysis of both tributyrin and trilinolein. The ester synthesis of unsaturated fatty acids with various alcohols catalyzed by the purified lipase in a nonaqueous medium or microaqueous system was also investigated. The esterification activity of the lipase increased with an increase of the carbon chain length in the alcohol. The synthesis rate of linoleic acid and oleyl alcohol was the highest with an optimal temperature and pH of $40^{\circ}C$ and 8.0, respectively. The water content and agitation also affected the esterification activity of the lipase.

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Effects of Pinus densiflora on soil chemical and microbial properties in Pb-contaminated forest soil

  • Kim, Sung-Hyun;Lee, In-Sook;Kang, Ho-Jeong
    • Journal of Ecology and Environment
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    • v.34 no.3
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    • pp.315-322
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    • 2011
  • We investigated the effect of Pb uptake by Pinus densiflora and the Pb fraction in forest soil. We also investigated the change in soil physicochemical characteristics, microbial activity, and root exudates of Pinus densiflora in Pb-contaminated soils. Three-year-old pine seedlings were exposed to 500 mg/kg Pb for 12 months. The metal fractions were measured using sequential extraction procedures. Additionally, factors that affect solubility (three soil enzyme activities and amino acids of root exudate compounds) were also determined. The results showed that Pb contamination significantly decreased enzyme activities due to soil characteristics. In addition, organic matter, nitrate content, and Pb concentration were time dependent. The results indicate that changes in the Pb fraction affected Pb uptake by pine trees due to an increase in the exchangeable Pb fraction. The concentrations of organic acids were higher in Pb-spiked soil than those in control soil. Higher rhizosphere concentrations of oxalic acid resulted in increased Pb uptake from the soil. These results suggest that pine trees can change soil properties using root exudates due to differences in the metal fraction.