• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1087-1093
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• 2005

The population diversity and seasonal changes of bacterial communities in rice soils were monitored using both culture-dependent approaches and molecular methods. The rice field plot consisted of twelve subplots planted with two genetically-modified (GM) rice and two non-GM rice plants in three replicates. The DGGE analysis revealed that the bacterial community structures of the twelve subplot soils were quite similar to each other in a given month, indicating that there were no significant differences in the structure of the soil microbial populations between GM rice and non-GM rice during the experiment. However, the DGGE profiles of June soil after a sudden flooding were quite different from those of the other months. The June profiles exhibited a few intense DNA bands, compared with the others, indicating that flooding of rice field stimulated selective growth of some indigenous microorganisms. Phylogenetic analysis of l6S rDNA sequences from cultivated isolates showed that, while the isolates obtained from April soil before flooding were relatively evenly distributed among diverse genera such as Arthrobacter, Streptomyces, Terrabacter, and Bacillus/Paenibacillus, those from June soil after flooding mostly belonged to the Arthrobacter species. Phylogenetic analysis of 16S rDNA sequences obtained from the soil by cloning showed that April, August, and October had more diverse microorganisms than June. The results of this study indicated that flooding of rice fields gave a significant impact on the indigenous microbial community structure; however, the initial structure was gradually recovered over time after a sudden flooding.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.656-660
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• 2013

Pseudomonas fluorescens strain CL0145A was discovered at the New York State Museum Field Research Laboratory as an effective agent against the environmentally destructive zebra mussel, which has contaminated US waters. Dried cells of the microbe are being commercialized as an environmentally friendly solution to the problem. We found that antibiotic activity against the Gram-positive bacterium Bacillus subtilis is produced and excreted by this strain. We have carried out studies to optimize production of the antibiotic. Studies were begun in a complex corn meal medium. Activity was found in both cells and culture supernates and was maximal after one day of fermentation. Static fermentation conditions were found to be superior to shaken culture. Production of extracellular antibiotic in complex medium was found to be dependent on the content of sucrose and enzyme-hydrolyzed casein. Indeed, production was greater in sucrose plus enzyme-hydrolyzed casein than in the complex medium. Of a large number of carbon sources studied as improvements over sucrose, the best was glycerol. An examination of nitrogen sources showed that production was improved by replacement of enzyme-hydrolyzed casein with soy hydrolysates. Production in the simple glycerol-Hy-Soy medium was not improved by addition of an inorganic salt mixture or by complex nitrogen sources, with the exception of malt extract. In an attempt to keep the medium more defined, we studied the effect of amino acids and vitamins as replacements for malt extract. Of 21 amino acids and 7 vitamins, we found tryptophan, glutamine, biotin, and riboflavin to be stimulatory. The final medium contained glycerol, Hy-Soy, tryptophan, glutamine, biotin, and riboflavin.

• Journal of Microbiology and Biotechnology
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• 제28권4호
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• pp.588-596
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• 2018

An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.

• Journal of Microbiology and Biotechnology
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• 제28권4호
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• pp.606-612
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• 2018

The enzyme xylose isomerase (E.C. 5.3.1.5, XI) is responsible for the conversion of an aldose to ketose, especially xylose to xylulose. Owing to the ability of XI to isomerize glucose to fructose, this enzyme is used in the food industry to prepare high-fructose corn syrup. Therefore, we studied the characteristics of XI from Anoxybacillus kamchatkensis G10, a thermophilic bacterium. First, the gene coding for XI (xylA) was inserted into the pET-21a(+) expression vector and the construct was transformed into the Escherichia coli competent cell BL21 (DE3). The expression of recombinant XI was induced in the absence of isopropyl-thio-${\beta}$-galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was $80^{\circ}C$ and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at $60^{\circ}C$. The optimum pH was found to be 7.5 in sodium phosphate buffer. Magnesium, manganese, and cobalt ions were found to increase the enzyme activity; manganese was the most effective. Additionally, recombinant XI was resistant to the presence of $Ca^{2+}$ and $Zn^{2+}$ ions. The kinetic properties, $K_m$ and $V_{max}$, were calculated as 81.44 mM and $2.237{\mu}mol/min/mg$, respectively. Through redundancy analysis, XI of A. kamchatkensis G10 was classified into a family containing type II XIs produced by the genera Geobacillus, Bacillus, and Thermotoga. These results suggested that the thermostable nature of XI of A. kamchatkensis G10 may be advantageous in industrial applications and food processing.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.227-233
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• 2015

Two recombinant arabinosyl hydrolases, α-_L-arabinofuranosidase from Geobacillus sp. KCTC 3012 (GAFase) and endo-(1,5)-α-_L-arabinanase from Bacillus licheniformis DSM13 (BlABNase), were overexpressed in Escherichia coli, and their synergistic modes of action against sugar beet (branched) arabinan were investigated. Whereas GAFase hydrolyzed 35.9% of _L-arabinose residues from sugar beet (branched) arabinan, endo-action of BlABNase released only 0.5% of _L-arabinose owing to its extremely low accessibility towards branched arabinan. Interestingly, the simultaneous treatment of GAFase and BlABNase could liberate approximately 91.2% of _L-arabinose from arabinan, which was significantly higher than any single exo-enzyme treatment (35.9%) or even stepwise exo- after endo-enzyme treatment (75.5%). Based on their unique modes of action, both exo- and endo-arabinosyl hydrolases can work in concert to catalyze the hydrolysis of arabinan to _L-arabinose. At the early stage in arabinan degradation, exo-acting GAFase could remove the terminal arabinose branches to generate debranched arabinan, which could be successively hydrolyzed into arabinooligosaccharides via the endo-action of BlABNase. At the final stage, the simultaneous actions of exo- and endo-hydrolases could synergistically accelerate the _L-arabinose production with high conversion yield.

• 멤브레인
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• 제20권2호
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• pp.127-134
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• 2010

연구는 음식물 쓰레기를 음식물 소멸기에서 미생물에 의해 발효하여 유기물을 감량화하고, 음식물 발효시 발생되는 가스를 냉각기에서 기체와 수분으로 분리하여 기체는 반응기로 다시 보내고 수분은 응축하여 침지식 MF 중공사 분리막이 장착된 MBR 시스템에 적용하여 처리하는 시스템에 관한 연구이다. (주)바이오하이테크에서 제작한 음식물 소멸기와 수처리 장치에 침지식 MF 중공사 모듈을 설치하여 90일간 H연구소 직원식당에서 발생되는 음식물 쓰레기를 연속 투입하여 처리하였다. 음식물 소멸기 초기 Seeding를 위하여 수분조절제로 미강, 왕겨, 톱밥을 305 kg 투입하였고 음식물은 운전기간동안 1,648 kg투입하였고 응축폐수는 1,600 L 발생되었다. 음식물 소멸기 운전 종료 후 배출된 발효 부산물은 386 kg으로 감량율은 약 80%로 조사되었다. 침지식 MF 중공사 분리막 모듈을 응축폐수의 MBR 시스템에 적용하여 유기물 처리한 결과 제거율은 각각 BOD 99.9%, COD 97.5%, SS 98.6%, T-N 54.6%, T-P 34.7%였으며, 총대장균은 100%가 제거되었다.

• 한국환경보건학회지
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• 제37권5호
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• pp.376-386
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• 2011

Objectives: The aim of this study was to evaluate the microbial hazards posed by food utensils and fixtures in food service operations at selected middle and high schools located in Seoul, Korea. Methods: We collected 200 samples of utensils and fixtures including cups, spoons/chopsticks, food trays and tables from five different schools in Seoul. Target microorganisms of this study were divided into two groups: total bacterial count and total coliform as indicators of microbial contamination and Bacillus cereus and Staphylococcus aureus as pathogens of food poisoning. We used selective media to quantify microbial concentration and 16S rRNA PCR assay for qualitative analysis. In addition, intensive interviews with nutritionists were conducted and observations were made to identify factors that may affect microbial contamination. Logistic regression analysis was employed to examine the relationship between the microbial concentration and operation characteristics of each operation. Results: The level of microbial concentration in school B and C were significantly lower than in school A, D and E (p<0.05). Some samples from school A, D and E showed over 3.4 log CFU/100 $cm^2$ (total bacterial count) and 1.0 log CFU/100 $cm^2$ (total coliform), which requires immediate hygienic action. The number of customers per staff member, periodicity of hygiene education for staff and daily operation time of sterilizers were also found to be important factors related with the microbial contamination of food service operations. Conclusions: These results suggested that not only a HACCP (Hazard Analysis and Critical Control Point) approach, but also efforts to assess internal risk factors within operations be needed to reduce the microbial contamination of food utensils and fixtures. This study is expected to provide preliminary data for assessing microbial hazards in food service operations.

• 한국환경과학회지
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• 제27권5호
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• pp.333-340
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• 2018

This study aimed to estimate the effect of inoculant application level on chemical composition and bacterial count of fermented chestnut meal (FCM), and its rumen fermentation characteristics. The inoculant contained Lactobacillus acidophilus ($1.2{\times}10^{10}cfu/g$), Bacillus subtilis ($2.1{\times}10^{10}cfu/g$), and Saccharomyces cerevisiae ($2.3{\times}10^{10}cfu/g$). The chestnut meal mixed with molasses, double distilled water, and inoculant at 1 kg, 3 g, 480 mL, and 20 mL ratio for the basal chestnut meal diet. The double distilled water from basal chestnut meal diet was substituted with bacterial inoculant at a level of 0 (Control), 20 (Medium), and 40 mL (High) in the experimental diets. The mixed experimental diets were incubated at $39^{\circ}C$ for 7, 14, and 21 days, respectively. On 7 days of FCM incubation, the contents of crude protein (CP) (quadratic, P=0.043) and neutral detergent fiber (quadratic, P=0.071) decreased by increases of inoculant application levels, whereas bacterial count (quadratic, P=0.065) and rumen $NH_3-N$ (linear, P=0.063) increased. By increases of inoculant application levels on 14 days of FCM incubation, the increases were found on dry matter (DM) (quadratic, P=0.085), CP (quadratic, P=0.059), acid detergent fiber (quadratic, P=0.056), in vitro DM digestibility (linear, P=0.002), rumen total volatile fatty acid (VFA) (linear, P=0.057), and rumen iso-butyrate (linear, P=0.054). However, the decreases were found on bacterial count (linear, P=0.002), propionate (linear, P=0.099), and butyrate (quadratic, P=0.082). On 21 days of FCM incubation, in vitro DM digestibility (linear, P=0.002) and total VFA (linear, P=0.001) increased by increases of inoculant application levels, whereas the contents of CP (quadratic, P=0.034) and neutral detergent fiber (quadratic, P=0.047) decreased. These results indicate that the FCM with a medium level of inoculant application and 14 of fermentation had beneficial effects by increasing DM digestibility and rumen total VFA content, without altering bacterial count.

• 한국환경과학회지
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• 제26권5호
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• pp.651-659
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• 2017

This study aimed to estimate the effects of replacing Mushroom By-Product (MBP) with Tofu By-Product (TBP) on the chemical composition, microbes, and rumen fermentation indices of Fermented Diets (FDs). The basal diet was formulated using MBP, TBP, rice bran, molasses, and inoculants. The MBP in the basal diet was replaced with TBP at 0, 5, and 10% on Dry Matter (DM) basis for the experimental diets. The experimental diets were fermented at $39^{\circ}C$ for 144 h. Chemical composition, pH, microbes, and rumen fermentation indices of the FDs were analyzed. With increasing TBP replacement, crude protein content of FDs increased (L, P < 0.001), whereas crude ash content decreased (L, P = 0.002). Lactic acid bacteria and Bacillus subtilis contents in the TBP-replaced FDs were higher than those in the control (P < 0.05), whereas pH level and mold count were lower (P < 0.05). With increasing TBP replacement, in vitro rumen digestibility of DM (L, P = 0.053) and neutral detergent fiber (L, P = 0.024) increased, wheres rumen pH changed (P = 0.026) quadratically. Rumen total volatile fatty acid (L, P = 0.001) and iso-butyrate contents (Q, P = 0.003) increased with increasing TBP replacement. In conclusion, this study indicates that the replacement of MBP with TBP could improve the quality of FD.

• 한국축산시설환경학회지
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• 제8권3호
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• pp.183-190
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• 2002

미생물제제를 이용한 돈분뇨 처리시스템(3N-System, 속성발효시스템) 운영에 있어 돼지에 급여하는 생균제가 비육돈의 발육 및 질병발생에 미치는 효과와 분뇨처리효과를 구명하기 위하여 실시한 시험결과를 요약하면 다음과 같다. 생균제 혼합급여 시험은 사료에 0.1% 씩 생균제와 항생제를 각각 첨가하여 처리구별로 30두씩 3처리 평균체중이 21.4$\pm$0.54kg인 육성비육돈 90두를 공시하여 시험을 실시하였다. 0.1% 생균제 (Bacillus toyoi spore) 급여구는 육성돈에서 출하시까지 급여하였으며, 0.1% 항생제 (kitasamycin and sulfamethazine) 첨가 급여구는 출하 1개월전 부터는 항생제 첨가한 사료를 급여하지 않았다. 일당 증체량과 사료효율은 처리간에 유의적인 차이를 나타내지 않았다(P >0.05). 도체성적에 있어 출하시 체중과 도체중은 항생제 첨가급여구가 다소 높게 나타났으나 처리간에 유의적인 차이는 없었다(P>0.05). 등지방 두께는 생균제 첨가 급여구가 다른 처리구에 비하여 낮게 나타나는 경향이었다(P<0.05). 도체등급별 출현두수는 A등급 출현율에서 생균제 첨가 급여구가 다소 높은 비율을 보였다. 육안 병변에 대한 모니터링 결과에서는 생균제 급여구가 병변발생율이 적게 나타났다. 시험돈사의 환경조사에서 처리간에 온도의 차이는 없었으나 최저온도에서는 항생제첨가 급여구가 다른 처리구에 비하여 유의적으로 높게 나타났다(P<0.01). 습도는 생균제 첨가급여구가 84.34$\pm$6.03으로 다른 처리구에 비하여 높게 나타났다(P<0.01). 시험돈사내의 돈분뇨의 성상변화(BOD, COD, SS, T-N, T-P 및 암모니아 농도)도 생균제첨가 급여구가 낮은 수치를 보였다.