• Asian-Australasian Journal of Animal Sciences
• /
• 제26권3호
• /
• pp.358-365
• /
• 2013

A series of in vitro studies were carried out to determine i) the effects of enzyme and formaldehyde treatment on the degradation characteristics of carbohydrate and protein sources and on the synchronicity of these processes, and ii) the effects of synchronizing carbohydrate and protein supply on rumen fermentation and microbial protein synthesis (MPS) in in vitro experiments. Untreated corn (C) and enzyme-treated corn (EC) were combined with soy bean meal with (ES) and without (S) enzyme treatment or formaldehyde treatment (FS). Six experimental feeds (CS, CES, CFS, ECS, ECES and ECFS) with different synchrony indices were prepared. Highly synchronous diets had the greatest dry matter (DM) digestibility when untreated corn was used. However, the degree of synchronicity did not influence DM digestibility when EC was mixed with various soybean meals. At time points of 12 h and 24 h of incubation, EC-containing diets showed lower ammonia-N concentrations than those of C-containing diets, irrespective of the degree of synchronicity, indicating that more efficient utilization of ammonia-N for MPS was achieved by ruminal microorganisms when EC was offered as a carbohydrate source. Within C-containing treatments, the purine base concentration increased as the diets were more synchronized. This effect was not observed when EC was offered. There were significant effects on VFA concentration of both C and S treatments and their interactions. Similar to purine concentrations, total VFA production and individual VFA concentration in the groups containing EC as an energy source was higher than those of other groups (CS, CES and CFS). The results of the present study suggested that the availability of energy or the protein source are the most limiting factors for rumen fermentation and MPS, rather than the degree of synchronicity.

• 한국식품영양과학회지
• /
• 제25권5호
• /
• pp.878-884
• /
• 1996

BailIus sp. IJ-3 균주는 균체외 효소로 대두단백 응고효소를 생산하고, 이 박테리아의 최적 효소생산에 대한 배지조성 및 배양조건을 검토하였다. 최대의 대두단백 응고효소를 생산하는 배지조성은 soymilk 20%(w/v), glucose 2.0%, yeast extract 5.0%, peptone 4.0% 및 potassium Phosphate, monobasic 1.0%였다. 초기 pH는 6.0이었고, $35^{\circ}C$ 배양에서 최대 효소활성을 보였다. 조정된 배지 조성에서 72시간 배양에 최대 효소활성을 나타내었으며, 기존배지 활성의 약 5.3배의 효소활성의 증가를 보였다. BaCilIus sp. IJ-3 균주효소는 7S globu1in에 대해 특이성을 갖고 있는 것으로 사료된다.

• Journal of Microbiology and Biotechnology
• /
• 제17권12호
• /
• pp.1983-1990
• /
• 2007

Antidiabetic effects of a novel microbial biopolymer (PGB) 1 excreted from new Enterobacter sp. BL-2 were tested in the db/db mice. The animals were divided into normal control, rosiglitazone (0.005%, wt/wt), low PGB1 (0.1%, wt/wt), and high PGB1 (0.25%, wt/wt) groups. After 5 weeks, the blood glucose levels of high PGB1 and rosiglitazone supplemented groups were significantly lower than those of the control group. In hepatic glucose metabolic enzyme activities, the glucokinase activities of PGB1 supplemented groups were significantly higher than the control group, whereas the PEPCK activities were significantly lower. The plasma insulin and hepatic glycogen levels of the low and high PGB1 supplemented groups were significantly higher compared with the control group. Specifically, the insulin and glycogen increases were dose-responsive to PGB1 supplement. PGB1 supplement did not affect the IPGTT and IPITT compared with the control group; however, rosiglitazone significantly improved IPITT. High PGB1 and rosiglitazone supplementation preserved the appearance of islets and insulin-positive cells in immunohistochemical photographs of the pancreas compared with the control group. These results demonstrated that high PGB1 (0.25% in the diet) supplementation seemingly contributes to preventing the onset and progression of type 2 diabetes by stimulating insulin secretion and enhancing the hepatic glucose metabolic enzyme activities.

• 한국지하수토양환경학회지:지하수토양환경
• /
• 제18권3호
• /
• pp.73-81
• /
• 2013

Various remediation methods have been applied to clean soils contaminated with pollutants. They remove contaminants from the soils by utilizing physicochemical, biological, and thermal processes and can satisfy soil remediation standards within a limited time; however, they also have an effect on the biological functions of soils by changing soil properties. In this study, changes of the biological properties of soils before and after treatment with three frequently used remediation methods-soil washing, land farming, and thermal desorption-were monitored to investigate the effects of remediation methods on soil biological functions. Total microbial number and soil enzyme activities, germination rate and growth of Brassica juncea, biomass change of Eisenia andrei were examined the effects on soil microorganisms, plant, and soil organisms, respectively. After soil washing, the germination rate of Brassica juncea increased but the above-ground growth and total microbial number decreased. Dehydrogenase activity, germination rate and above-ground growth increased in both land farming and thermal desorption treated soil. Although the growth of Eisenia andrei in thermal desorption treated soil was higher than any other treatment, it was still lower than that in non-contaminated soil. These results show that the remediation processes used to clean contaminated soil also affect soil biological functions. To utilize the cleaned soil for healthy and more value-added purposes, soil improvement and process development are needed.

• Journal of Microbiology and Biotechnology
• /
• 제24권6호
• /
• pp.843-853
• /
• 2014

In the context of artificial groundwater recharge, a reactive soil column at pilot-scale (4.5 m depth and 3 m in diameter) fed by treated wastewater was designed to evaluate soil filtration ability. Here, as a part of this project, the impact of treated wastewater filtration on soil bacterial communities and the soil's biological ability for wastewater treatment as well as the relevance of the use of multi-bioindicators were studied as a function of depth and time. Biomass; bacterial 16S rRNA gene diversity fingerprints; potential nitrifying, denitrifying, and sulfate-reducing activities; and functional gene (amo, nir, nar, and dsr) detection were analyzed to highlight the real and potential microbial activity and diversity within the soil column. These bioindicators show that topsoil (0 to 20 cm depth) was the more active and the more impacted by treated wastewater filtration. Nitrification was the main activity in the pilot. No sulfate-reducing activity or dsr genes were detected during the first 6 months of wastewater application. Denitrification was also absent, but genes of denitrifying bacteria were detected, suggesting that the denitrifying process may occur rapidly if adequate chemical conditions are favored within the soil column. Results also underline that a dry period (20 days without any wastewater supply) significantly impacted soil bacterial diversity, leading to a decrease of enzyme activities and biomass. Finally, our work shows that treated wastewater filtration leads to a modification of the bacterial genetic and functional structures in topsoil.

• Asian-Australasian Journal of Animal Sciences
• /
• 제15권12호
• /
• pp.1725-1731
• /
• 2002

The distribution and activities of hydrolytic enzymes (cellulolyti, hemicellulolytic,pectinolytic and others) in the rumen compartments of Hereford bulls fed 100% alfalfa hay based diets were evaluated. The alfalfa proportion in the diet was gradually increased for two weeks. Whole rumen contents were processed into four fractions: Rumen contents including both the liquid and solid fractions were homogenized and centrifuged, and the supernatant was assayed for enzymes located in whole rumen contents (WRE); rumen contents were centrifuged and the supernatant was assayed for enzymes located in rumen fluids (RFE); feed particles in rumen contents were separated manually, washed with buffer, resuspended in an equal volume of buffer, homogenized and centrifuged and supernatant was assayed for enzymes associated with feed particles (FAE); and rumen microbial cell fraction was separated by centrifugation, suspended in an equal volume of buffer, sonicated and centrifuged, and the supernatant was assayed for enzymes bound with microbial cells (CBE). It was found that polysaccharide-degrading proteins such as $\beta$-1,4-D-endoglucanase, $\beta$-1,4-D-exoglucanase, xylanase and pectinase enzymes were located mainly with the cell bound (CBE) fraction. However, $\beta$-D-glucosidase, $\beta$-D-fucosidase, acetylesterase, and $\alpha$-L-arabinofuranosidase were located in the rumen fluids (RFE) fraction. Protease activity distributions were 37.7, 22.1 and 40.2%, and amylase activity distributions were 51.6, 18.2 and 30.2% for the RFE, FAE and CBE fractions, respectively. These results indicated that protease is located mainly in rumen fluid and with microbial cells, whereas amylase was located mainly in the rumen fluid.

• 생태와환경
• /
• 제46권2호
• /
• pp.276-288
• /
• 2013

본 연구는 질소 시비에 의해 증가된 토양 질소가 식물의 성장 및 식물체의 화학적 조성에 미치는 영향과 이로 인한 분해에서의 변화를 확인하고자 야외성장실험과 분해실험을 진행하였다. 온실에서 질소 시비구와 비시구 토양에 각각 벼를 재배하였으며 식물이 성숙한 뒤 수확하여 C, N, lignin, cellulose 함량을 측정하였다. 대조구와 질소 처리구 토양에서 재배된 식물의 개체 당 평균 건중량은 각각 0.70 g, 1.32 g로 질소 시비에 의해 1.9배 증가하였다. 식물체의 N 및 C 함량은 질소 시비에 의해 증가하였고 lignin, C/N, lignin/N, cellulose/N은 감소하였다. 이후, 수확된 식물의 지상부는 microcosm 분해실험에 이용되었으며, 분해 식물체에서 건중량의 변화, microbial biomass C와 microbial biomass N, 그리고 dehydrogenase와 urease 활성을 측정하고, 분해과정 중 발생하는 $CO_2$의 양을 정량하였다. 대조구 토양에서 분해시킨 대조구 식물체와 질소 처리구 식물체, 그리고 질소를 처리한 토양에서 분해시킨 질소 처리구 식물체의 잔존량은 각각 초기 건중량의 53.0%, 47.1%, 53.6%를 나타내었다. 질소 시비는 식물체에서 N 함량을 높이고 C/N 및 lignin/N을 낮추어 식물체의 분해를 촉진하였으나, 분해 과정에서의 토양 질소처리는 분해를 억제하였다. 질소 시비에 의해 토양에서 microbial biomass C와 dehydrogenase 활성은 감소하였고, 반면에 microbial biomass N과 urease 활성은 증가하였다. 분해 중 발생한 $CO_2$의 양은 30일 이후부터 질소 시비에 의해 감소하였다. 분해 식물체에서 측정된 microbial biomass C는 질소 처리에 의해 초기에 증가하였으나 이후 저해되는 양상을 나타냈으며 microbial biomass N은 유의한 차이를 보이지 않았다. 질소 시비에 의해 분해 식물체에서 dehydrogenase 활성은 저해되었으며 urease는 분해 초기에 가장 높은 활성을 보였으나 분해 후기에 현저한 감소를 나타냈다. 본 실험에서 질소 시비는 식물의 성장을 증가시키고 식물체의 N 함량을 높여 화학적 조성의 변화를 일으키며 분해율을 증가시키나 분해 단계에서 질소의 시비는 미생물의 활성을 억제시켜 분해를 저해하는 결과를 나타내었다.

• Journal of Microbiology and Biotechnology
• /
• 제22권8호
• /
• pp.1133-1140
• /
• 2012

Ribonucleic acid interference (RNAi) inhibits the expression of target genes in a sequence-specific manner, and shows potential for gene knockdown in filamentous fungi, in which the locus-specific gene knockout occurs in low frequency. In this study, the function of the repressor of cellulase expression I (ACEI) was verified in Trichoderma koningii (T. koningii) YC01 through RNAi, and ace1-silenced strains with improved cellulase productivity were obtained. An expression cassette that transcribed the interfering double-stranded RNA (dsRNA) of ace1 was constructed and transformed into T. koningii, and the transformants, in which the expression of ace1 was successfully silenced, were selected. As a result of the ace1 gene silencing, the expression levels of the main cellulase and xylanase genes were elevated, and the enhanced production of total proteins, cellulase, and xylanase was observed in the cultivation. In addition, the down-regulation of ace1 resulted in an increasing expression of xyr1, but no clear variation in the expression of cre1, which suggested that ACEI acted as a repressor of the xyr1 transcription, but was not involved in the regulation of the cre1 expression. The results of this work indicate that ace1 is a valid target gene for enhancing enzyme production in T. koningii, and RNAi is an appropriate tool for improving the properties of industrial fungi.

• 한국미생물·생명공학회지
• /
• 제27권4호
• /
• pp.311-319
• /
• 1999

Leaves of Aloe vera Linne and bloods of domestic animal were composted in a soild state fermentation reactor (SSFR) by using microbial additive including a bulking and moisture controlling agent. From solid-culture of microbial additive, 10 species of bacteria and 10 species of fungi were isolated and, their enzyme activities including amylase, carboxy methyl cellulase CMCase, lipase and protease were detected. Optimum fermentation conditions of Aloe leaves and domestic animal bloods in SSFR were obtained from the studies of response surface analysis employing microbial additive content, initial moisture content, and fermentation temperature as the independent variables. The optimum conditions for SSFR using Aloe leaves were obtained at 9.45$\pm$73%(w/w) of microbial additives, 62.73$\pm$4.54%(w/w) of initial moisture content and 55.32$\pm$3.14$^{\circ}C$ of fermentation temperature while those for SSFR using domestic animal bloods were obtained at 10.25$\pm$2.04%, 58.68$\pm$4.97% and 57.85$\pm$5.$65^{\circ}C$, respectively. Composting process in SSFR was initially proceeded through fermentation and solid materials were decomposed within 24 hours by maintaining higher moisture level, and maturing and drying steps are followed later. After the fermentation step, the concentrations of solid phase inorganic components were increased while that of organic components were decreased. Also, concentrations of total organic carbon(TOC), peptides, amino acids, polysaccharides, and low fatty acids in water extracts were increased. As fermentation in composting process depends on initial C/N ratios in water extracts of two samples were increased because of increased water-soluble TOC. From these results, it was revealed that solid state fermentation reactor using microbial additives can be used in composting process of organic wastes with broad C/N ratio.

• 한국토양비료학회지
• /
• 제49권2호
• /
• pp.144-156
• /
• 2016

Soil is a dynamic biological system, in which it is difficult to determine the composition of microbial communities. Knowledge of microbial diversity and function in soils are limited because of the taxonomic and methodological limitations associated with studying the organisms. In this review, approaches to measure microbial diversity in soil were discussed. Research on soil microbes can be categorized as structural diversity, functional diversity and genetic diversity studies, and these include cultivation based and cultivation independent methods. Cultivation independent technique to evaluate soil structural diversity include different techniques such as Phospholipid Fatty Acids (PLFA) and Fatty Acid Methyl Ester (FAME) analysis. Carbon source utilization pattern of soil microorganisms by Community Level Physiological Profiling (CLPP), catabolic responses by Substrate Induced Respiration technique (SIR) and soil microbial enzyme activities are discussed. Genetic diversity of soil microorganisms using molecular techniques such as 16S rDNA analysis Denaturing Gradient Gel Electrophoresis (DGGE) / Temperature Gradient Gel Electrophoresis (TGGE), Terminal Restriction Fragment Length Polymorphism (T-RFLP), Single Strand Conformation Polymorphism (SSCP), Restriction Fragment Length Polymorphism (RFLP) / Amplified Ribosomal DNA Restriction Analysis (ARDRA) and Ribosomal Intergenic Spacer Analysis (RISA) are also discussed. The chapter ends with a final conclusion on the advantages and disadvantages of different techniques and advances in molecular techniques to study the soil microbial diversity.