• Title/Summary/Keyword: Bacillus protease

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Antagonistic Activities of Bacillus spp. Strains Isolated from Tidal Flat Sediment Towards Anthracnose Pathogens Colletotrichum acutatum and C. gloeosporioides in South Korea

  • Han, Joon-Hee;Shim, Hongsik;Shin, Jong-Hwan;Kim, Kyoung Su
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.165-175
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    • 2015
  • Anthracnose is a fungal disease caused by Colletotrichum species that is detrimental to numerous plant species. Anthracnose control with fungicides has both human health and environmental safety implications. Despite increasing public concerns, fungicide use will continue in the absence of viable alternatives. There have been relatively less efforts to search antagonistic bacteria from mudflats harboring microbial diversity. A total of 420 bacterial strains were isolated from mudflats near the western sea of South Korea. Five bacterial strains, LB01, LB14, HM03, HM17, and LB15, were characterized as having antifungal properties in the presence of C. acutatum and C. gloeosporioides. The three Bacillus atrophaeus strains, LB14, HM03, and HM17, produced large quantities of chitinase and protease enzymes, whereas the B. amyloliquefaciens strain LB01 produced protease and cellulase enzymes. Two important antagonistic traits, siderophore production and solubilization of insoluble phosphate, were observed in the three B. atrophaeus strains. Analyses of disease suppression revealed that LB14 was most effective for suppressing the incidence of anthracnose symptoms on pepper fruits. LB14 produced antagonistic compounds and suppressed conidial germination of C. acutatum and C. gloeosporioides. The results from the present study will provide a basis for developing a reliable alternative to fungicides for anthracnose control.

Effect of feeding fermented soybean meal on broiler chickens' performance: a meta-analysis

  • Irawan, Agung;Ratriyanto, Adi;Respati, Adib Norma;Ningsih, Niati;Fitriastuti, Rahma;Suprayogi, Wara Pratitis Sabar;Hadi, Rendi Fathoni;Setyono, Wahyu;Akhirini, Novi;Jayanegara, Anuraga
    • Animal Bioscience
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    • v.35 no.12
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    • pp.1881-1891
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    • 2022
  • Objective: The present study aimed to quantify the effects of fermented soybean meal (FSBM) on broiler chickens' performance by employing a meta-analysis approach. Methods: A total of 16 studies were included in the database after being systematically selected using a PRISMA protocol. Hedges' g effect size was used to quantify pooled standardized mean difference (SMD) using random-effects models at 95% confidence intervals (95% CI). Publication bias among studies was computed with Egger's test and visualized using funnel plots. Results: Results indicated that dietary FSBM inclusion increased final body weight (BW) (SMD = 0.586, 95% CI: 0.221 to 0.951, p = 0.002) of broiler chickens, particularly in starter period (SMD = 0.691, 95% CL: 0.149 to 1.233, p = 0.013) while in the finisher period, the effect was weaker (SMD = 0.509, 95% CI: 0.015 to 1.004, p = 0.043). Average daily gain (ADG), feed intake (FI), and feed conversion ratio (FCR) were not affected with FSBM inclusion when compared to control. Subgroup analysis revealed that FI increased in starter period (SMD = 0.582, 95% CI: 0.037 to 1.128, p = 0.036). When considering types of microorganism as moderating variables in the subgroup analysis, we found that Aspergillus oryzae, mixed probiotics+bromelain protease, Bacillus subtilis, and Lactobacillus bacteria significantly increased ADG and FI (p<0.01). Additionally, either Bacillus subtilis+protease or Bacillus subtilis alone decreased FCR (p<0.001). However, meta-regression analysis showed that levels of FSBM inclusion had no effects on final BW (p = 0.502), ADG (p = 0.588), FI (p = 0.861), and FCR (p = 0.462). Conclusion: Substituting SBM in broiler chickens' diet with FSBM improved BW of broiler chickens, especially in the starter period whereas the effects on ADG, FI, and FCR were mostly dependent on microbial strains used for fermentation.

Evaluation of Rhizobacterial Isolates for Their Antagonistic Effects against Various Phytopathogenic Fungi (식물 근권에서 분리한 미생물의 식물병원성 진균에 대한 길항효과 검정)

  • Kim, Yun Seok;Kim, Sang woo;Lamsal, Kabir;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.44 no.1
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    • pp.36-47
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    • 2016
  • This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.

Control of Red Pepper Anthracnose Using Bacillus subtilis YGB36, a Plant Growth Promoting Rhizobacterium (식물생장촉진근권세균 Bacillus subtilis YGB36을 이용한 고추 탄저병의 생물학적 방제)

  • Lee, Yong Yoon;Lee, Younmi;Kim, Young Soo;Kim, Hyun Sup;Jeon, Yongho
    • Research in Plant Disease
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    • v.26 no.1
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    • pp.8-18
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    • 2020
  • Red pepper, one of the major economic crops in Korea, is being affected by anthracnose disease caused by Colletotrichum acutatum. To control this disease, an antagonistic bacterial strain, Bacillus subtilis YGB36 identified by 16S rDNA sequencing, physiological and biochemical analyses is used as a biological control agent. In vitro screening revealed that the strain YGB36 possess strong antifungal activity against the pathogen Cylindrocarpon destructans. The strain exhibited cellulase, protease, amylase, siderophore production and phosphate solubility. In vitro conidial germination of C. acutatum was most drastically inhibited by YGB36 cell suspensions (106 cfu/ml) or culture filtrate. Development of anthracnose symptoms was reduced on detached immature green pepper fruits by treatment with cell suspensions, and its control value was recorded as 65.7%. The YGB36 bacterial suspension treatment enhanced the germination rate of red pepper seeds and promoted root development and growth under greenhouse conditions. The in vitro screening of fungicide and insecticide sensitivity test against YGB36 revealed that the bacterial growth was not affected by any of the insecticides, and 11 fungicides out of 21 used. Collectively, our results clearly suggest that the strain YGB36 is considered as one of the potential biocontrol agents against anthracnose disease in red pepper.

Effects of Inoculation of Bacillus subtilis Cells on the Fermentation of Korean Traditional Soy Paste(Doenjang) (메주 제조 시 Bacillus subtilis의 첨가가 재래식 된장의 발효에 미치는 영향)

  • Rhee, Chang-Ho;Kim, Won-Chan;Rhee, In-Koo;Park, Heui-Dong
    • Food Science and Preservation
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    • v.15 no.4
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    • pp.598-605
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    • 2008
  • Korean traditional soy paste(Doenjang) was fermented for 90 days using, as a starter, a Meju prepared by inoculation of Bacillus subtilis cells. Changes in physiochemical and functional properties were investigated during fermentation. Amylase and protease activities increased and showed maximal levels(4.11 and 7.75 units/mL, respectively) after 60 days of fermentation. Both enzyme activities then fell. Inhibitory activities of the soy paste against tyrosinase and angiotensin converting enzyme(ACE) increased during the fermentation period. Antimutagenic activities during fermentation were determined using the S. enterica serovar Typhimurium TA100 and TA98 analysis system. No significant differences in activity were observed in soy pastes prepared with or without B. subtilis. Antimutagenic activities against the activities of N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and 4-nitro-O-phenylenediamine (NPD) increased and reached 70.33% and 60.22%, respectively, after 90 days of fermentation, as assessed using the tester strain TA100. Against the actions of NPD and 4-nitroquinoline-1-oxide(NQO), antimutagenic activities also increased during fermentation and reached 50.84% and 47.01%, respectively, as assessed using the tester strain TA98. The genistin content was much higher(187.48 g/g) in soy paste prepared using B. subtilis inoculation than the level(31.30 g/g) seen in soy paste prepared without bacterial inoculation, although the contents of daidzein and genistein were slightly reduced under such circumstances.

Effects of Bacillus SW1-1 coated diets on innate immunity and disease resistance of olive flounder Paralichthys olivaceus against Edwardsiella tarda infection

  • Kim, Min-Gi;Gunathilaka, Buddi E.;Lee, Sungho;Kim, Youjeong;Lee, Kyeong-Jun
    • Fisheries and Aquatic Sciences
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    • v.25 no.4
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    • pp.243-249
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    • 2022
  • Bacillus SW1-1 is a probiotic isolated from shrimp intestines. We investigated the effects of Bacillus SW1-1 coated diets on the growth, feed utilization, innate immunity, hematological parameters and resistance to Edwardsiella tarda in olive flounder (Paralichthys olivaceus). A commercial diet was used as the control (AP0) and two other diets were prepared by coating 0.25% (AP25) or 0.50% (AP50) probiotic powder which contains 1.0 × 107 CFU/g Bacillus SW1-1. Four replicate groups of olive flounder (153 ± 2 g) were fed one of the diets for 12 weeks. Growth performance and feed utilization of the fish were not significantly affected by the dietary Bacillus SW1-1. After the challenge with E. tarda, AP50 group showed significantly higher survival than AP0 and AP25 groups. Innate immunity and anti-oxidant capacity of the fish were not significantly affected after the feeding trial. However, after the E. tarda challenge, the innate immune parameters (immunoglobulin, lysozyme and anti-protease) were significantly improved in fish fed AP25 and AP50 diets compared to those in fish fed AP0 diet. After the challenge test, significantly lower glucose level was observed in AP50 group compared to AP0 group. These results indicate that dietary supplementation of Bacillus SW1-1 could increase the disease resistance of olive flounder against E. tarda infection. The optimum coating levels of Bacillus SW1-1 needs to be further elucidated.

Isolation, Identification, and Characterization of Bacillus strains from the Traditional Korean Soybean-fermented Food, Chungkookjang

  • Joo, Myeong-Hoon;Hur, Sung-Ho;Han, Yong-Soo;Kim, Ji-Yeon
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.202-210
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    • 2007
  • A total of 45 bacterial strains were isolated from the traditional Korean soybean-fermented food, Chungkookjang. Among these strains, seven strains were selected and identified based on morphological, physiological, and biochemical characteristics, as well as phylogenetic analysis using 16S rDNA sequences. All strains were Gram-positive, aerobic, motile, oxidase-positive, rod-shaped, and endospore-forming bacteria, and produced extracellular enzymes such as amylase, cellulase, lipase, protease, and xylanase. The isolates were grown in the presence of 0-11% (w/v) NaCl. Growth was optimal at pH 6-9 and at temperatures of $30-45^{\circ}C$. According to VITEK automicrobic system tests and supplementary tests, the isolates were similar to several species of the genus Bacillus. The phylogenetic analysis of seven bacterial strains based on comparisons of 16S rDNA sequences, revealed that the strains were closely related to Bacillus species. The identification of strains that produced surfactin was also carried out, based on PCR screening of the sfp gene. Among the seven isolated strains, six yielded a surfactin-positive result with PCR.

Purification and Identification of pretense from Bacillus sp. HB-5 and its application of cosmetic product (Bacillus sp. HB-5균주가 생산하는 단백질 분해 효소의 분리. 정제 및 화장품에의 응용)

  • 이범천;윤은정;이동환;표형배
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.26 no.1
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    • pp.107-124
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    • 2000
  • A bacterial strain No. HB-5, which was capable of producing a pretense in the culture conditions, was isolated from the soil . The pretense was purified from cultural filtrate of Bacillus sp. HB-5 by membrane ultrafiltration and DEAE- cellulose chromatography, gel filtration on Sephadex G-100. The molecular weight was estimated to be 60k4a. The optimal pH and temperature for the activity of the purified pretense pH were 11 and 5$0^{\circ}C$ , respectively. The enzyme was stable within a pH range 8-12 and up to 6$0^{\circ}C$ . The enzyme activity was highly inhibited by PMSF at 1mM. The proteolytic actions of pretense and papain on human epidermis keratins which are major protein impurities on the skin, were compared. The bacterial pretense degraded more effectively than papain. Product containing 2% protease exhibited 21% increase on the skin coloration index. These results suggest that cosmetic product containing pretense produced by Bacillus sp. HB-5 could remove the adherent keratin layer and then make a softer skin.

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Identification of Enteric Bacteria from Nephila clavata (한국산 무당거미(Nephila clavata)에서 분리한 장내 세균의 동정)

  • 문은영;오현우;맹필재;배경숙
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.1-8
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    • 2001
  • Spiders are carnivores that prey upon insects and other small arthropods through digestion of food outside the body. Although spider poison may contain proteolytic enzymes, these are thought to play an insignificant role in actual digestion. The source of active proteolytic enzymes can be either the digestive tract cells of spider, or natural microbial flora in the digestive tract of spider. In this study, digestive tracts from the spider, Nephila clavata, were screened for bacteria that have protease or lipase activity. A total of $10^3-10^5$ CFU was recovered from a spider and more than 90% of them showed protease and lipase activity respectively. Of the microbial isolates, 63.3% showed protease or lipase activity, and 50% of these showed both protease and lipase activity. Some of the isolates were characterized using a battery of chemical, phenotypic and genotypic methods. Eleven Gram negative bacteriaa (Acinetobacter calcoaceticus, A. haemolyticus, Alcaligenes faecalis, Cedecea davisae, C. neteri, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas fluorescens, Serratia marcescens, Stenotrophomonas maltophilia, Suttonella indologenes) and 11 Gram positive bacteria (Bacillus cereus, B. coagulans, B. pasteurii, B. thuringiensis, Cellulomonas flavigena, Corynebacterium martruchotii, Enterococcus durans, E. faecalis, Micrococcus luteus, Staphylococcus hominis, S. sciuri) were identified.

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Isolation of Bacillus subtilis CK-2 Hydrolysing Various Organic Materials (다양한 유기물을 분해하는 Bacillus subtilis CK-2의 분리)

  • Kim, Chul-Ho;Lee, Sang-Hyup
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1716-1720
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    • 2011
  • A bacterium hydrolysing various organic materials including cellulose, protein, starch and lipid was isolated. The isolate was identified as Bacillus subtilis, and named Bacillus subtilis CK-2 in this paper. This bacterium showed optimal growth at $40\sim45^{\circ}C$, pH 6~9, and 0~3% of NaCl. B. subtilis CK-2 seemed to synthesis highly active autolysin. The hydrolytic enzymes produced by B. subtilis CK-2 were primary enzymes because extracellular enzyme activities varied similarly to the growth curve. The hydrolytic enzymes seemed to be stable at basic pH conditions. From these results, B. subtilis CK-2 was found to bea useful bacterial agent for composting, or for use in feed-production waste in agriculture, fishery, forest materials, livestock farming, and food.