• Title/Summary/Keyword: Bacillus protease

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The Quality of Doenjang (Soybean Paste) Manufactured with Bacillus brevis (Bacillus brevis 로 제조한 된장의 품질)

  • 양성호;최명락;지원대;정영건;김종규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.6
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    • pp.980-985
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    • 1994
  • We investigated the quality of soybean paste(Doenjang) fermented by BAcillus brevis. The results obtained were as follows : soybean paste fermented by Bacillus brevis had alkaline pH and yellow ochre color. Dextrinizing activity was about 98 D.P. unit from 5th to 25th day of fermentation at 3$0^{\circ}C$ and after that day somewhat decreased . Saccharifying activity was respectively 6.1, 7.2, 6.8, 6.4 S.P. unit on 5, 15, 25 and 35th day of fermentation. Protease activity suddenly increased after 15th day of fermentation and was 250, 275, 299 unit on15, 25, 35 th day of fermentation , respectively. The most abundant free amino acid was found to be glutamic acid (561.8mg%) in soybean paste fermented by Bacillus brevis. In case of free sugar and non-volatile organic acid, fructose and oxalic acid showed highest content of 10.25mg% on 25th day and 12.20mg% on 15th day. The contents of free amino acids, free sugars, organic acids in soybean paste fermented by B.brevis were most abundant after 25 days of fermentation and this results were similar to that of traditional soybean paste. The odor of soybean paste fermented by Bacillus brevis was improved to be a nice soybean paste odor on 25th day of fermentation. However, sensory evaluation value of the taste of it decreased after 10 th day of fermentation.

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Selection and Characterization of Bacillus licheniformis MH48 for the Biocontrol of Pine Wood Nematode (Bursaphelenchus xylophilus) (소나무재선충 생물학적 방제를 위한 Bacillus licheniformis MH48의 선발 및 특성 규명)

  • Jeong, Min-Hae;Yang, Seo-Young;Lee, Yong-Sung;Ahn, Young-Sang;Park, Yun-Serk;Han, Hye-rim;Kim, Kil-Yong
    • Journal of Korean Society of Forest Science
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    • v.104 no.3
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    • pp.512-518
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    • 2015
  • Pine wilt disease (PWD) caused by pine wood nematode, Bursaphelenchus xylophilus, has become the most serious threat to pine trees in Korea. This study was subjected to investigate effective biological control agent against PWD. To select nematocidal bacteria against PWD, Bacillus licheniformis MH48 was selected among five bacteria due to its high nematocidal potential. B. licheniformis MH48 was tested for cell growth and protease activity to evaluate its nematicidal potential. In the B. licheniformis MH48, cell numbers were highest three days after incubation, while protease activity was highest after seven days. In the effect of different concentrations of B. licheniformis MH48 culture broth against B. xylophilus, 20% concentration of culture broth showed approximately 80% of pine wood nematode mortality compared to the control. Especially, pine wood nematode's cuticle layers were degraded two days after treatment of B. licheniformis MH48 culture broth. The present study suggests that B. licheniformis MH48 can be one of the potential biocontrol candidates against pine wood nematode due to its ability to produce protease.

Quality characteristics of buckwheat Soksungjang manufactured by Bacillus subtilis HJ18-4 (Bacillus subtilis HJ18-4를 이용하여 제조한 메밀 속성장의 품질특성)

  • Park, Na Young;Lee, Sun Young;Kim, Ji Yeun;Choi, Hye Sun
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.699-704
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    • 2013
  • Buckwheat Soksungjang (BS) is a bealmijang manufactured with buckwheat and soybeans. We manufactured BS using Bacillus subtilis HJ18-4 (HJ18-4), which has high enzyme activities and antibacterial effects. HJ18-4 was inoculated in a different process during the BS manufacturing, which was the meju-making time (Treat 1), and the salt water time was added (Treat 2). The physiochemical and microbial characteristics of the BS were analyzed. As a result, the total aerobic counts (7~8 log CFU/mL) in the BS increased after 15 days of fermentation. Especially, Treat 1 showed higher total aerobic counts and amino-type nitrogen (65.38~202.52 mg%) than Treat 2. During the BS fermentation, the reduction of the sugar contents and the enzyme (protease and amylase) activities decreased. In the relative quantitative expression level of PlcR, Treat 1 did not show toxin gene expressions at the end of the fermentation on Day 23. Treat 1 showed suitable B. cereus physiochemical quality characteristics and inhibition effects. When the modified-form type of fermented soybean paste was manufactured with a single starter, it could not reproduce the natural fermentation quality. These results suggest that the addition of a starter (HJ18-4) in the Meju manufacturing process could enhance the quality characteristics of the manufactured BS via natural fermentation and by suppressing B. cereus.

A Thermostable Protease Produced from Bacillus sp. DF 218 (Bacillus sp. DF218이 생산하는 내열성 단백질 분해효소)

  • Lee, Joung-Hee;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.105-110
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    • 2004
  • Microorganism (strain DF 218) producing thermostable pretense was isolated from Korean soil and compost. It was Gram-positive, rod-shaped, aerobic, and spore-forming with yellowish white colony color, Temperature range for growth at pH 6.5 was $30-65^{\circ}C$, with optimum growth at $60^{\circ}C$. pH range for growth at $60^{\circ}C$ was 5-7 with optimum of 6.5, which indicates strain DF 218 to be thermophilic. The 16S rDNA sequence of strain DF 218 had 95% sequence similarity with that of Bacillus flexus. Based on physiological properties and phylogenetic analysis, we proposed the isolated strain as Bacillus sp. DF 218. Pretense was produced aerobically at $60^{\circ}C$ for 32 hr in a medium (pH 6.5) containing 1% each trypton, glucose, and NaCl. Its molecular weight was estimated as 61 kDa, with optimum temperature and pH of $60^{\circ}C$ and 7.5, respectively.

Isolation and Characterization of an Alkaline Protease Produced by Bacillus subtilis JK-1 (알칼리성 Pretease를 생산하는 Bacillus subtilis JK-1의 분리 동정 및 효소 특성)

  • Kim, Ji-Yeon
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.331-336
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    • 2007
  • A bacterium producing the alkaline pretense was isolated from Chungkookjoug, and was identified as Bacillus subtilis JK-1 based on morphological, physiological and biochemical characteristics, as well as phylogenetic analysis using 16S rRNA gene sequence. The optimum pH and temperature of the pretense activity were pH 9.0 and $55^{\circ}C$, respectively. This enzyme was stable at the temperatures $40{\sim}80^{\circ}C$. The maximum alkaline pretense production was obtained when 1.0% (w/v) xylose, 1.0% (w/v) yeast extract and 0.3% (w/v) $CuSO_4$ were used as carbon source, nitrogen source and mineral source. Under the optimal condition, growth of the isolate was reached at stationary phase after 12 hr followed by incubation, the alkaline pretense production reached a maximum level with $16{\sim}20$ hr cultivation.

Properties of Cheonggukjang Fermented with Bacillus Strains with High Fibrinolytic Activities

  • Jeong, Woo-Ju;Lee, Ae-Ran;Chun, Ji-Yeon;Cha, Jae-Ho;Song, Young-Sun;Kim, Jeong-Hwan
    • Preventive Nutrition and Food Science
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    • v.14 no.3
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    • pp.252-259
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    • 2009
  • We previously isolated Bacillus strains with high fibrinolytic activities (FAs) from cheonggukjang prepared by traditional ways. To test their potential as starters for cheonggukjang, soybean was fermented for 72 hr at $37^{\circ}C$ with each isolate and a control lab strain: B. subtilis CH3-25 (BS3-25), B. amyloliquefaciens CH51 (BA51), B. amyloliquefaciens CH86-1 (BA86-1), and B. subtilis 168 (BS168, control, lab strain). Viable cell numbers of all cheonggukjang samples rapidly increased and reached about $10^9$ CFU/g after 6 hr. During 72 hr, the initial pH of 6.3 rapidly increased to 8.1$\sim$8.2 for cheonggukjang fermented with BS3-25 or BA86-1, and 7.3 for those with BA51 or BS168. FAs and protease activities (acid, neutral, and alkaline) rapidly increased in cheonggukjang fermented with BS3-25, BA51, or BA86-1 during the first 12 hr. On the other hand, those of cheonggukjang fermented with BS168 slightly increased during the first 36 hr. There were significant changes in acid and neutral protease activities in cheonggukjang fermented with BA51 or BA86-1 during the 24 hr. Rapid increases of $\beta$-glucosidase activity corresponded well with rapid increases of $\alpha$-amylase and $\alpha$-galactosidase activities in addition to increases in antioxidant activities and the TPCs (total phenolic contents). The highest increase in the TPCs was observed in cheonggukjang fermented with BA86-1 while the least was that fermented with BS168.

Purification and Characterization of a New Peptidase, Bacillopeptidase DJ-2, Having Fibrinolytic Activity: Produced by Bacillus sp. DJ-2 from Doen-Jang

  • CHOI, NACK-SHICK;YOO, KI-HYUN;HAHM, JEUNG-HO;YOON, KAB-SEOG;CHANG, KYU-TAE;HYUN, BYUNG-HWA;PIL, JAE-MAENG;KIM, SEUNG-HO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.72-79
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    • 2005
  • A new Bacillus peptidase, bacillopeptidase DJ-2 (bpDJ-2), with molecular mass of 42 kDa and isoelectric point (pI) of 3.5- 3.7, was purified to homogeneity from Bacillus sp. DJ-2 isolated from Doen-Jang, a traditional Korean soybean fermented food. The enzyme was identified as an extracellular serine fibrinolytic protease. The optimal conditions for the reaction were pH 9.0 and $60^{\circ}C$. The first 18 amino acid residues of the N-terminal amino acid sequence of bpDJ-2 were TDGVEWNVDQIDAPKAW, which is identical to that of bacillopeptidase F (bpf). However, based on their Nterminal amino acid sequence, molecular size, and pI, it is different from that of bpf and extracellular 90 kDa. The whole (2,541 bp, full-bpDJ-2) and mature (1,956 bp, mature-bpDJ-2) genes were cloned, and its nucleotide sequence and deduced amino acid sequence were determined. The expressed proteins, full-bpDJ-2 and mature-bpDJ-2, were detected on SDSPAGE at expected sizes of 92 and 68 kDa, respectively.

A Study on the Extraction of Alkaline Protease from Bacillus licheniformis Fermentation Broth using Reverse Micelle (역미셀을 이용한 Bacillus licheniformis 발효액으로 부터 알카리성 단백질 분해효소의 추출에 관한 연구)

  • 권성필;구윤모홍성안
    • KSBB Journal
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    • v.6 no.1
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    • pp.105-109
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    • 1991
  • In separating alkaline protease from the bacteria (Bacillus licheniformis) fermentation broth using reverse micelle, effects of various factors;ionic strength, pH and surfactant concentration, on separation efficiency were studied. KCl controls the ionic strength. The lower KCl concentration was in the feed solution, the more protein and activity was recovered. The higher KCl concentration was in the stripping solution, the more protein and activity was recovered. Using sodium-di-2-ethylhexyl sulfosuccinate(Aerosol-OT or AOT) as a surfactant, the higher AOT concentration in the solvent, the more activity and protein were recovered. 0.1N NaOH and IN HCl were used to adjust pH. Maximum recovery of protein mass and activity were obtained at feed solution of pH 5.3. Maximum activity was recovered at stripping solution of pH 7.5

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Production of Bioactive Components and Anti-Oxidative Activity of Soybean Grit Fermented with Bacillus subtilis HA according to Fermentation Time (Bacillus subtilis HA를 이용한 soybean grit의 고체발효 기간에 따른 생리활성물질 생산 및 항산화 효과)

  • Kim, Ji-Eun;Lee, Sam-Pin
    • Korean Journal of Food Science and Technology
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    • v.41 no.2
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    • pp.179-185
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    • 2009
  • Soybean grits, fortified with various bioactive components, were produced by solid-state fermentation using Bacillus subtilis HA. ${\alpha}$-Amylase activity gradually increased during fermentation over 5 days. Fibrinolytic and protease activities were highest in the soybean grits fermented for 7 days. The grits fermented for 5 days also showed the highest tyrosine content, indicating a higher peptide content. Peptides of low molecular weight (below 3,000 daltons) and browning pigments increased with increasing fermentation time. The fermented soybean grits showed higher contents of total phenolic compounds, to approximately 18 mg/g. DPPH free radical scavenging effects were higher in the soybean grits fermented for 3 days. Also, ABTS radical scavenging effects were greater in the fermented grits compared to the unfermented grits. Overall, the soybean grits fermented by solid-state fermentation for 5 days showed enhanced production of bioactive compounds and greater antioxidant properties.

Antifungal Activity of Bacillus polyfermenticus CJ6 Isolated from Meju (메주로부터 분리한 Bacillus polyfermenticus CJ6의 항진균 활성)

  • Jung, Ji-Hye;Chang, Hae-Choon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.4
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    • pp.509-516
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    • 2009
  • The CJ6 bacterial strain, which possesses strong antifungal activity, was isolated from meju and identified as Bacillus polyfermenticus based on Gram staining, biochemical properties, and 16S rRNA gene sequencing. B. polyfermenticus CJ6 showed antimicrobial activity against the various pathogenic molds, yeasts, and bacteria. Antifungal activity from B. polyfermenticus CJ6 was reduced after 24 hr at $70^{\circ}C$ but antifungal activity was not completely destroyed. The antifungal activity was stable in the pH range of $3.0{\sim}9.0$, and inactivated by proteinase K, protease, and ${\alpha}$-chymotrypsin, which indicate its proteinaceous nature. The apparent molecular masses of the partially purified antifungal compound, as indicated by using the direct detection method in Tricine-SDS-PAGE, was approximately 1.4 kDa.