• Preventive Nutrition and Food Science
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• 제9권1호
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• pp.14-20
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• 2004

Bacillus strains capable of producing fibrinolytic enzyme were isolated from traditional fermented Korean soybean paste and Japanese fermented soybean (Natto). Among the 16 strains, a selected Bacillus sp. was identified as bacillus firmus, with 80.7％ homology, by API kit analysis. Seed starter or B. firmus NA-1 was prepared with 5％ soymilk prepared from micronized soybean powder. To produce fibrinolytic enzyme by B. firmus NA-1 the liquid culture was performed with NB broth (pH 7.0) fortified with 1％ galactose, 0.1% tryptone, and 0.5% $K_2$HPO$_4$, by shaking with 180 rpm at 37$^{\circ}C$. Fibrinolytic enzyme activity reached the highest value at 7.8 unit/mL (plasmin unit) after fermentation for 72 hr. The crude fibrinolytic enzyme showed higher relative activity in the range of pH 7.0∼9.0. The activity of crude fibrinolytic enzyme was well maintained even after concentration by the vacuum evaporation at 5$0^{\circ}C$ for 1 hr.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.13-14
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• 2001

Doenjang (Korean fermented soybean paste) is one of important fermented foods in Korea. Doenjang has been traditionally manufactured from meju which is fermented rectangular shape of crushed cooked soybeans. The main microorganisms involved for meju fermentation are Bacillus subtilis and molds such as Rizopus sp., Mucor sp. and Aspergillus sp. We have already reported that Doenjang is free from mycotoxin, especially, aflatoxin B$_1$contamination during the manufacturing process of the Deonjang.(omitted)

• 미생물학회지
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• 제49권1호
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• pp.38-45
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• 2013

본 연구에서는 연안갯벌 갯지렁이(Perinereis aibuhitensis)에서 분리한 광염성 미생물, Bacillus sp. EBW4의 특성과 다양한 환경조건에서 유기물 분해능을 분석하였다. 16S rRNA 염기서열에 기초하여 동정한 결과, 균주 EBW4는 Bacillus hemicentroti $JSM076093^T$와 98.3%, Bacillus hwajinponensis SW-$72^T$와 97.96% 그리고 Bacillus algicoa $KMM3737^T$와 96.28%의 상동성을 보였다. EBW4의 생장 가능한 온도 범위는 $4-40^{\circ}C$, 염분도 범위는 0-17%, pH 범위는 5-9로 나타나 EBW4는 광염성 균주로 밝혀졌다. EBW4의 세포막을 구성하는 주요 지방산으로는 anteiso $C_{15:0}$, iso $C_{16:0}$, anteiso $C_{17:0}$, iso $C_{14:0}$ 등으로 각각 48.2, 12.1, 11.6, 9.4% 비율로 나타났다. EBW4는 탄수화물, 단백질, 지방 등 다양한 고분자 유기물을 분해할 수 있는 DNase, amylase, protease, lipase 등의 효소 활성뿐만 아니라, alkaline phosphatase, esterase (C4), leucine arylamidase 그리고 ${\alpha}$-chymotrypsin 효소활성도 가지고 있었다. 다양한 염분 농도 조건에서 합성폐수를 이용한 실험에서 EBW4은 조사한 모든 범위의 염분 조건에서도 유기물 분해능이 우수하였다.

• 한국식품영양과학회지
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• 제45권3호
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• pp.460-465
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• 2016

본 연구에서는 Bacillus sp.를 이용하여 감자슬라이스를 발효시킨 후 성분을 분석하고, 감자 칩으로 가공하여 색도와 아크릴아마이드 함량을 알아보았다. B. licheniformis, B. methylotrophicus, B. subtilis 각각을 $7.5{\times}10^7{\sim}2.7{\times}10^8CFU/mL$ 접종하여 6시간 동안 발효한 결과 균수의 증가는 보이지 않는 유도기였음에도 불구하고 총당 함량은 감소하였으며, 환원당 함량 변화에 영향을 미쳤다. 감자 칩의 색도는 대조구에 비하여 실험구의 L 값이 증가하였으며, a 값과 b 값은 감소하여 감자 칩으로 적합한 색도를 나타내었다. 발효 감자 칩의 아크릴아마이드 함량은 평균 약 94% 감소하였다.

• 한국식품저장유통학회지
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• 제18권2호
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• pp.171-177
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• 2011

더덕의 유용성을 입증하기 위하여 더덕의 기능성과 더덕이 첨가된 청국장의 품질을 조사하였다. 70% 에탄올에서 추출한 더덕 추출물의 DPPH radical 제거능이 물 추출물보다 높았다. Salmonella typhimurium TA 98을 이용하여 직접 변이원에 대한 항변이원성 시험을 하였다. 200, 1,000, 2000, 3,000, 4,000 ug/plate의 에탄올 더덕 추출물에 대한 항돌연변이원율이 5.75, 31.38, 34.75, 53.50과 83.75%였으며 물 추출물보다 2배 이상 높았다. 청국장 제조에 더덕 함량을 5, 10, 15 및 20% (w/w)로 달리하여 첨가 제조한 청국장에 대하여 이화학적 및 관능적 검사 등의 품질에 미치는 영향을 조사하였다. 청국장 제조에 사용한 균주는 재래식 메주에서 ${\alpha}$-amyalse, ${\beta}$-amylase 및 산성 protease 활성이 우수한 것으로 선정된 Bacillus sp. B-3이다. 숙성 중 아미노태질소 함량을 조사한 결과 10% (w/w) 더덕 함유 청국장이 다른 처리구에 비해 가장 많이 생성되고 있었다. 한편, 더덕 함량이 15% (w/w) 이상 함유된 더덕 청국장에서는 아미노태질소 함량이 감소하는 경향을 보여 더덕 함량이 된장 중의 Bacillus sp. B-3의 생육에 영향을 미치는 것으로 나타났다. 색도에서는 더덕 함량이 높을수록 L-값이 감소하는 것으로 나타났다. 더덕 10% (w/w)가 첨가된 청국장이 기호도 측면에서 다른 처리구에 비해 우수한 것으로 나타났다.

• 환경위생공학
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• 제13권2호
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• pp.34-39
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• 1998

For more convenient and rapidly isolation of Bacillus thuringiensis subsp. israelensis(Bti), 1) heat treatment spore forming bacteria, 2) growth in enrichment culture media for Bacillus sp. and 3) selection of bacteria producing a lecithinase for Bacillus thuringiensis subsp. israelensis, were performed. Spore forming bacteria were counted 4.8 $\times $ 10$^{8}$cells/g soil on NAPGCY media, 9.2 $\times $ 10$^{7}$cells/g soil on NA media, and 3.6 $\times $ 10$^{8}$cells/g soil on NAAC media, respectively. Bacteria producing only a lecithinase were reached at 25.2% on medium contained egg york, bacteria only producing a delta-endotoxin were reached at 23.2% by phase contrast microscope, and bacteria producing a lecithinase & a delta-endotoxin simultaneously were reached at 13.7%. Bacillus thuringiensis which producing a lecithinase and a delta-endotoxin simultaneously among bacteria producing a lecithinase, were reached at 56.5%; A half of Bacillus thuringiensis was produced a delta-endotoxin, but not produced a lecithinase. Among 8 isolates of Bacillus thuringiensis, two strain of Bti which has a mosquito-cidal toxin, were detected by PCR using a specific primer of $\delta $-endotoxin gene from Bti.

• 한국미생물·생명공학회지
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• 제23권5호
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• pp.506-512
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• 1995

A cell aggregation factor produced by Aspergillus sp. LAM 94-142 was purified and partially characterized. The factor was purified about 15 folds from culture broth by IRA 420 and IRC 120 treatment, 1% NaCl added acetone precipitation, and Sepharose 4B column chromatography with overall yield of 48%. It was heteropolysaccharide consisted of mannose, arabinose, and glucose with a molar ratio, 31:17:2, and its molecular weight was estimated to be about 900,000 daltons by Sepharodse 4B gel filtration method. The optimum pH and temperature was 8 and 40$\circ$C, respectively. The factor was stable in pH range of 3-9 and at 100$\circ$C for 90 min. The cell aggregation activity of the factor was inhibited by the addition of Hg$^{2+}$, Fe$^{2+}$, Cu$^{2+}$, and some polypeptides such as milk casein or hemoglobin. The factor aggregated Bacillus subtilis, B. macerans, B. turingiensis, E. coli, Peudomonas aeruginosa, P. fluorescens, P. malophilia, and weakly aggregated Staphylococcus sp., Sarcina lutea, P. putida and Cryptococcus neoformnans, but it didn't aggregate various strains of Candida sp. and Saccharomyces sp.

• Journal of Microbiology and Biotechnology
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• 제29권2호
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• pp.274-282
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• 2019

Mercury-resistant ($Hg^R$) bacteria were isolated from heavy metal polluted wastewater and soil collected near to tanneries of district Kasur, Pakistan. Bacterial isolates AZ-1, AZ-2 and AZ-3 showed resistance up to $40{\mu}g/ml$ against mercuric chloride ($HgCl_2$). 16S rDNA ribotyping and phylogenetic analysis were performed for the characterization of selected isolates as Bacillus sp. AZ-1 (KT270477), Bacillus cereus AZ-2 (KT270478) and Bacillus cereus AZ-3 (KT270479). Phylogenetic relationship on the basis of merA nucleotide sequence confirmed 51-100% homology with the corresponding region of the merA gene of already reported mercury-resistant Gram-positive bacteria. The merE gene involved in the transportation of elemental mercury ($Hg^0$) via cell membrane was cloned for the first time into pHLV vector and transformed in overexpressed C43(DE3) E. coli cells. The recombinant plasmid (pHLMerE) was expressed and the native MerE protein was obtained after thrombin cleavage by size exclusion chromatography (SEC). The purification of fusion/recombinant and native protein MerE by Ni-NTA column, dialysis and fast protein liquid chromatography (FPLC/SEC) involved unfolding/refolding techniques. A small-scale reservoir of wastewater containing $30{\mu}g/ml$ of $HgCl_2$ was designed to check the detoxification ability of selected strains. It resulted in 83% detoxification of mercury by B. cereus AZ-2 and B. cereus AZ-3, and 76% detoxification by Bacillus sp. AZ-1 respectively (p < 0.05).

• Applied Biological Chemistry
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• 제47권4호
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• pp.379-383
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• 2004

Bacillus sp. 유래 ${\beta}-mannanase$의 brown copra meal에 대한 기질 특이성을 규명하기 위하여 정제효소 150 ml를 0.5% 기질에 $50^{\circ}C$, 24 hrs 가수분해후 TLC, FACE로 비교 검토한 결과 2종류의 galactosyl mannooligosaccharide로 구성되어 1차 activated carbon column chromatography을 이용해 250 ml/hr 유속으로 tube당 50ml씩 ethanol $0{\sim}30%$ linear gradient로 당을 분리하였다. Activated carbon column chromatography에 의한 당용액 0.2 ml와 5% phenol 0.2 ml를 가하여 혼합 후 $Conc.H_2SO_4$ 1 ml를 가하여 혼합한 후 20분간 방치하여 490 nm로 흡광도를 측정하여 TLC로 pattern을 검토한 후 $Gal^3Man_4$ 및 DP 7의 galactosyl manooligosaccharide의 fraction을 회수하여 2차 activated carbon column chromatography를 이용해 1차와 동일한 조전에서 분리 회수하여 중합도 5는 $Gal^3Man_4(63-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$로 동정되었고, 중합도 7은 현재 동정중에 있다. Bifidobacterium속 균주(B. longum, B. bifidum)에 대한 생육활성을 비교 검토하기 위하여 MRS media에서 탄소원을 dextrose대신에 조제된 $Gal^3Man_4$를 첨가후 측정한 결과 $Gal^3Man_4$이 첨가되지 않은 MRS broth에 비해 생육촉진 활성을 보였다. 특히 B. longum에 대해서는 $Gal^3Man_4$를 dextrose대체 탄소원으로 처리시 10배의 생육활성이 증가하였다.

• Journal of Microbiology and Biotechnology
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• 제3권3호
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• pp.146-155
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• 1993

A DNA fragment from an alkali-tolerent Bacillus sp., conferring strong promoter activity, was subcloned into the promoter probe plasmid pPL703 and the nucleotide sequence of this promoter region was determined. The sequence analysis suggested that this highly efficient promoter region containing the complex clustered promoters comprised three kinds of promoters (P1, P2 and P3), which are transcribed by $\sigma^B (formerly \sigma^{37}), \sigma^E(formerly \sigma^{29}) and \sigma^A (formerly \sigma^{43})$ RNA polymerase holoenzymes which play major rules at the onset of endospore formation, during sporulation and at the vegetative phase of growth, respectively. S1 nuclease mapping experiments showed that all three promoters had staggered transcription initiation points. The results of chloramphenicol acetyltransferase assay after the subcloning experiments also indicated that the expression of these clustered promoters was correlated with the programs of growth and endospore development. Promoter P1, P2 and P3 were preceded by 75% AT, 79% AT and 81% AT regions, respectively, and a partial deletion of AT-rich region prevented transcription from promoter P1 in vivo. Two sets of 5 -AGTGTT-3 sequences and inverted repeat sequences located around the promoter P1 were speculated as the possible cis acting sites for the catabolite repression in B. subtilis. In vivo transcripts from these sequence regions may be able to form a secondary structure, however, the possibility that a regulatory protein induced by the excess amount of glucose could be bound to such a domain for crucial action remains to be determined.