• Title/Summary/Keyword: cellulase activities

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Isolation and Characterization of Yam-Putrefactive Psychrotrophic Bacteria from Rotted Yam (생마 저온부패 원인세균의 분리 및 부패균의 특성)

  • Ryu Hee-Young;Kim Young-Sook;Park Sang-Jo;Lee Bong-Ho;Kwon Soon-Tae;Sohn Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.109-114
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    • 2006
  • Yam has been recognized as healthy food due to its various biological activities, such as anti-obesity, antimicrobial, anticancer and immuno-stimulation activities, and its consumption has been increased during last decades. In this study, to investigate low-temperature, long-term storage of yam and to develop processed yam products, yam-putrefactive psychrotrophic bacteria were isolated from rotted yam and identified based on BBL identification system, fatty acid analysis in cell membrane and 16S rDNA sequence analysis. The putrefaction activity of isolated thirteen bacteria was evaluated using yam-slices (NaOCl-treated, autoclaved yam and without treatment), and YAM-10 and YAM-12 were identified as major psychrotrophic putrefactive bacteria. Both YAM-10 (Pseudomonas cepacia) and YAM-12 (Pseudomonas rhodesiae) bacteria grew well at 4$\sim$12$^{\circ}C$ and showed strong activity of polymer degrading enzymes, especially amylase, carboxy methyl cellulase and xylanase, at 20$^{\circ}C$. But they failed to grow at acidic pH (<5) or alkaline pH (>10). Our results suggested that the control of psychrotrophic Pseudomonas sp. by pH change and inhibition of polymer degrading enzymes, such as amy-lase, are necessary to long-term storage of yam.

Physiological and Biochemical Characterization of Bacillus spp. from Polychaete, Perinereis aibuhitensis (갯지렁이(Perinereis aibuhitensis)에서 분리한 Bacillus spp.의 생리생화학적 특성 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Journal of Life Science
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    • v.23 no.3
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    • pp.415-425
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    • 2013
  • This study compared the characteristics of five Bacillus strains capable of aerobic and anaerobic growth, CBW3, CBW4, CBW9, CBW14 and EBW10. They were isolated and selected from a polychaete, Perinereis aibuhitensis, which is known as a good degrader of organic compounds in marine wetland. Based on a 16S rRNA sequence, CBW3 and CBW14 were found to share more than 99.8% similarity with B. nanhaiensis, B. arsenicus and B. barbaricus. CBW4, CBW9 and EBW10 shared 92.7%, 99.8%, and 99.8% similarity with B. anthracis, B. algicoa and B. thuringiensis, respectively. The temperature, salinity, and pH ranges of the cell growth of the Bacillus strains were $4-45^{\circ}C$, 0-17%, and pH 5-pH 9, respectively. All Bacillus strains were found to exhibit enzyme activities for the degradation of casein and starch. Notably, strain EBW10 exhibited the enzyme activities for all the tested macromolecules, DNA, casein, starch, cellulose, and four kinds of Tweens, which suggests the possibility that it had protease, amylase, cellulose, and lipase. All five Bacillus strains had alkaline phosphatase activities, and the strains CBW3, CBW4, and EBW10 also had acid phospatase. Strains CBW3 and EBW10 exhibited the enzyme activities both for esterase (C4) and esterase lipase (C8). The analysis of fatty acids revealed that in all strains, major fatty acids were anteiso $C_{15:0}$ and iso $C_{15:0}$.

Biological Control of Fusarium Stalk Rot of Maize Using Bacillus spp. (Bacillus spp.를 이용한 옥수수 밑둥썩음병의 생물학적 방제)

  • Han, Joon-Hee;Park, Gi-Chang;Kim, Joon-Oh;Kim, Kyoung Su
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.280-289
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    • 2015
  • Maize (Zea mays L.) is an economically important crop in worldwide. While the consumption of the maize is steadily increasing, the yield is decreasing due to continuous mono-cultivation and infection of soil-borne fungal pathogens such as Fusarium species. Recently, stalk rot disease in maize, caused by F. subglutinans and F. temperatum has been reported in Korea. In this study, we isolated bacterial isolates in rhizosphere soil of maize and subsequently tested for antagonistic activities against F. subglutinans and F. temperatum. A total of 1,357 bacterial strains were isolated from rhizosphere. Among them three bacterial isolates (GC02, GC07, GC08) were selected, based on antagonistic effects against Fusarium species. The isolates GC02 and GC07 were most efficient in inhibiting the mycelium growth of the pathogens. The three isolates GC02, GC07 and GC08 were identified as Bacillus methylotrophicus, B. amyloliquefaciens and B. thuringiensis using 16S rRNA sequence analysis, respectively. GC02 and GC07 bacterial suspensions were able to suppress over 80% conidial germination of the pathogens. GC02, GC07 and GC08 were capable of producing large quantities of protease enzymes, whereas the isolates GC07 and GC08 produced cellulase enzymes. The isolates GC02 and GC07 were more efficient in phosphate solubilization and siderophore production than GC08. Analysis of disease suppression revealed that GC07 was most effective in suppressing the disease development of stalk rot. It was also found that B. methylotrophicus GC02 and B. amyloliquefaciens GC07 have an ability to inhibit the growth of other plant pathogenic fungi. This study indicated B. methylotrophicus GC02 and B. amyloliquefaciens GC07 has potential for being used for the development of a biological control agent.

Identification of a new marine bacterium Ruegeria sp. 50C-3 isolated from seawater of Uljin in Korea and production of thermostable enzymes (대한민국 울진 연안 해양에서 분리한 해양 미생물 Ruegeria sp. 50C-3의 동정 및 내열성 효소 생산)

  • Chi, Won-Jae;Kim, Jong-Hee;Park, Jae-Seon;Hong, Soon-Kwang
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.344-351
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    • 2016
  • A marine bacterium, designated as strain 50C-3, was isolated from a seawater sample collected from the East Sea of South Korea. The strain is a Gram-negative, aerobic, yellow colored polar-flagellated bacterium that grows at $20-50^{\circ}C$ and pH 5.5-8.5. Optimal growth occurred at $40-50^{\circ}C$, at pH 6.5-7.5, and in the presence of 2% (w/v) NaCl. Based on 16S rRNA gene sequence similarity, the isolate was considered to represent a member of the genus Ruegeria. The result of this analysis showed that strain 50C-3 shared 99.4% and 96.98% sequence similarity with Ruegeria intermedia CC-GIMAT-$2^T$ and Ruegeria lacuscaerulensis ITI-$1157^T$, respectively. Furthermore, strain 50C-3 showed clear differences from related strains in terms of several characteristics such as motility, carbon utilization, enzyme production, etc. The DNA G+C content was 66.7 mol%. Chemotaxonomic analysis indicated ubiquinone-10 (Q-10) as the predominant respiratory quinone. Based on phenotypic, chemotaxonomic, and phylogenetic characteristics, the isolate represents a novel variant of the Ruegeria intermedia CC-GIMAT-$2^T$, for which we named Ruegeria sp. 50C-3 (KCTC23890=DSM25519). Strain 50C-3 did not produce cellulase and agarase, but produced alkaline phosphatase, ${\alpha}$-galactosidase, and ${\beta}$-galactosidase. The three enzymes showed stable activities even at $50^{\circ}C$ and thus regarded as thermostable enzymes. Especially, the ${\beta}$-galactosidase activity enhanced by 1.9 times at $50^{\circ}C$ than that at $37^{\circ}C$, which may be very useful for industrial application.

Ecological Studies on the Transition of Sheath Blight of Rice in Korea (한국(韓國)에서의 벼 잎집무늬마름병 발생변동(發生變動)에 관(關)한 생태학적(生態學的) 연구(硏究))

  • Yu, Seung-hun
    • Korean Journal of Agricultural Science
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    • v.4 no.2
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    • pp.283-316
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    • 1977
  • In an attempt to obtain a basic information to develop an effective integrated system of controlling sheath blight of rice in Korea, the transition of this disease, the variation of cultural characters and pathogenicity of the pathogen, environmental conditions affecting the disease outbreak and varietal resistance have been investigated. 1. Rice sheath blight which has been minor disease in the past was widely spread, especially since 1971. This disease has promptly spread all over the country and infected 65.2% of total rice growing area in 1976. Various factors are considered to be related to such transition of this disease. Above all, increace of application of nitrogenous fertilizer, early season and earlier cultivation of rice, introduction of more susceptible "Tongil" varieties etc. must be important factors influencing the outbreak of this disease. 2. Great variations in cultural characteristics-such as mycelial growth rate, color of the medium, amount of the aerial mycelium, shape and color of the sclerotia- and in the pathogenicity of isolates of the pathogen, Thanatephorus cucumeris Dank were observed. The optimum temperature for mycelial growth also varied with isolates, from $25^{\circ}C$ to $30^{\circ}C$. There were not necessarily any correlation between curtural characteristics and pathogenicity of isolates of Thanatephorus cucumens. 3. Mycelial grow th of isolates of Thanatephorus cucumens on the PDA medium were correlated with the air temperatures of the region where the isolates were collected. The isolates from the regions with high temperature grew well on PDA medium at $35^{\circ}C$ than those from the region with low temperature, on the other hand, the isolates from the regions with the low temperature grew well on the same medium at $12^{\circ}C$ than those from the regions with high temperature. 4. Pectin polygalacturonase (PG) and cellulase (Cx) were most active on the 3rd day after inoculation on the leaves of rice plant with Thanatephorus cucumeris, whereas pectin methylestrase (PE) was most active on the 4th day after inoculation. Relationship between the activities of PE of isolates and the strength of pathogenicity of isolates was obtained, but PG and cellulase activities were not correlated with pathogenicity of isolates. 5. The tolerence of sclerotia from in-vitro culture to low temperature varied with their water content, the dried cultural sclerotia were more tolerent than wet ones, Dried cultural sclerotia maintained almost 100% germinability for 45 days at $-20^{\circ}C$, whereas wet sclerotia lost viability at $-5^{\circ}C$. The germination ratio of the sclerotia after overwintering changed from 18% to 70% according to the water content of the test paddy fields and the ratio was low in wet paddy condition. 6. To investigate the host range of this fungi in and near paddy field, 17 weeds were inoculated with fungi. The lesions of sheath blight disease was obserbed on Sagittaria trifolia L., Echinochloa crusgalli P. Beauv., Monochoria vaginal is Presl, Polygonum Hydropiper L., Eclipta prostrata L., Digitaria sanguinalis Scapoli. 7. When the level of nitrogen applied was doubled over standard level, total nitrogen content in rice sheath increased, ami when silicate was applied, starch content in rice sheath decreased, inducing the rice plants more susceptible to sheath blight disease. Increased dressing of potash ferilizer reduced the incidence of sheat blight disease. 8. The percentage of infected stems in the early period increased more in the narrow hill plot than in the wide hill plot, but in the late period this tendency was inversed; the percentage of infected stems as well as severity in the wide hill plot increased more compared to the narrow hill plot, and the disease severity in the one plant per hill plot was also low. The number of stems in the wide hill plot was more than the number of stems in the narrow hill plot. This indicates that the microclimate, such as the relative humidity, in the narrow hill plot was more favorable for the development of this disease. 9. There was a high negative correlation between the disease severity of varieties to the sheath blight and the maturity of the varieties, that is, the early varieties were more susceptible than the late ones, and much-tillering varieties usually showed more infection than less tillering varieties. 10. No relationship was obtained between the percentage of infected stems in the early period and the severity after heading, whereas a distinct relationship was obtained between former and latter after Aug. 10.

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Isolation and biochemical characterization of acid tolerance xylanase producing Bacteria, Bacillus sp. GJY from city park soil (도심공원으로부터 산내성 xylanase를 생산하는 박테리아 분리 및 효소학적 특성)

  • Jang, Min-Young;Park, Hwa Rang;Lee, Chong Gyu;Choo, Gab-Chul;Cho, Hyun Seo;Park, Sam-Bong;Oh, Ki-Cheol;Kim, Bong-Gyu
    • Journal of Applied Biological Chemistry
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    • v.60 no.1
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    • pp.79-86
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    • 2017
  • Microbes in forest are very important due to not only to enhance soil fertility but also maintain a healthy ecosystem by supplying the energy available to living organisms by producing various kinds of enzymes related to degradation of lignocellulosic biomass. In order to isolate a lignocellulosic biomass degrading bacterial strain from the Jurassic park located in Gyeongnam National University of Science and Technology, We used the Luria-Bertani-Carboxymethyl cellulose (CMC) agar trypan blue method containing 0.4 % carboxymethyl cellulose and 0.01 % trypan blue. As a result, we isolated a bacterial strain showing both activity on the CMC and xylan. To identify the isolated strain, 16S rRNA sequencing and API kit analysis were used. The isolated strain turned out to belong to Bacillus species and then named Bacillus sp. GJY. In the CMC zymogram analysis, it showed that one active band of about 28kDa in size is present. Xylan zymogram analysis also showed to have one active band of about 25kDa in size. The optimal growth temperature of Bacillus sp. GJY was $37^{\circ}C$. The maximal activities of CMCase and xylanase were 12 hour after incubation. The optimal pH and temperature for CMCase were 5.0 and $40^{\circ}C$, respectively, whereas the optimal pH and temperature for xylanase was 4.0 and $40^{\circ}C$. Both activities for CMCase and xylanase showed to be thermally stable at 40and $50^{\circ}C$, while both activities rapidly decreased at over $60^{\circ}C$.

Development of Optimal Culture Media for the Stable Production of Mushroom (버섯의 안정적 생산을 위한 최적배지의 개발)

  • Gal, Sang-Wan;Lee, Sang-Won
    • Applied Biological Chemistry
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    • v.45 no.2
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    • pp.71-76
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    • 2002
  • Several antagonistic bacteria, SD-1, 4, 10, 11, 14, 15, and 16, which have strong CMCase and amylase activities, were isolated from the fermented mushroom media. Among them, SD-1, 10, 11, and 15 have strong antibacterial activities against the mushroom pathogenic bacteria Pseudomonas sp., and SD-1, 10, 11, 14, and 16 have strong antifungal activities against the mushroom pathogenic fungi, Trichoderma sp. SD-14, 15, and 16 did not inhibit the growth of mushroom Pleurotus eryngii ASI-2302, and Pleurotus ostreatus ASI-2042 and ASI-2180. When the culture broth mixture of the seven bacterial strains was applied to the mushroom media, the growths of pathogens, Pseudomonas sp. and Trichoderma sp., were inhibited.

Changes in Biological Qualities of Soy Grits Cheonggukjang by Fermentation with β-Glucosidase-Producing Bacillus Strains (β-Glucosidase 활성이 있는 균주 Bacillus Strains를 접종해 제조한 Soy Grits 청국장의 품질 특성)

  • Lee, Kyung Ha;Choi, Hye Sun;Hwang, Kyung A;Song, Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.5
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    • pp.702-710
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    • 2016
  • The bioactivity of soy grits cheonggukjang was enhanced by fermentation using three ${\beta}$-glucosidase-producing Bacillus subtilis strains (HJ 18-9, HJ 25-8, and HJ 18-9+HJ 25-8) for 48 h at $37^{\circ}C$. The results indicate that protease, cellulase, and a-amylase activities significantly increased (P<0.05) with increasing fermentation time. In addition, the amino-type nitrogen content of B. subtilis-fermented soy grits cheonggukjang increased to 91.0~168.0 mg% after 48 h of fermentation. Among the isoflavones in soy grits cheonggukjang, contents of ${\beta}$-glucosides or acetyl-glucosides were reduced while aglycone content increased upon fermentation. In particular, soy grits cheonggukjang fermented with B. subtilis HJ18-9 and HJ25-8 showed the largest increases in aglycone content compared to complex treatment. These results provide useful information for development starter (single and complex) as well as for production of high quality fermented soybean food.

Isolation and Identification of Hydrolytic Enzyme-producing Bacteria from Spent Mushroom Substrate (버섯부산물유래 가수분해효소분비 박테리아의 분리 및 동정)

  • Kim, Young-Il;Jeong, Se-Hyung;Seok, Joon-Sang;Yang, Si-Yong;Huh, Jung-Won;Kwak, Wan-Sup
    • Journal of Animal Science and Technology
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    • v.50 no.5
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    • pp.713-720
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    • 2008
  • This study was conducted to isolate and identify xylanase- and cellulase-producing thermophilic bacteria from stacked spent mushroom substrates and to determine the optimal medium conditions for their growth. Bacteria with the highest xylanase and CMCase activities were strain 3 and 201-7. Both of them were identified as Bacillus spp. and named Bacillus subtilis KU3 and Bacillus subtilis KU201-7. The optimal medium condition of Bacillus subtilis KU3 was obtained when 3%(w/v) of yeast extract and 1%(w/v) of maltose were used as nitrogen and carbon sources, respectively. That of Bacillus subtilis KU201-7 was obtained when 0.5%(w/v) of yeast extract and 0.5%(w/v) of CMC were used as nitrogen and carbon sources, respectively.

Increase of Bioactive Flavonoid Aglycone Extractable from Korean Citrus Peel by Carbohydrate-Hydrol-ysing Enzymes (당 분해효소를 이용한 감귤 Flavonoid 무배당체 함량의 증가)

  • Ahn Soon-Cheol;Kim Min-Soo;Lee Sun-Hi;Kang Ju-Hyung;Kim Bo-Hye;Oh Won-Keun;Kim Bo-Yeon;Ahn Jong-Seog
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.288-294
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    • 2005
  • Flavonoid compounds show several biological activities and generally exist in the forms of glycones linking sugar moiety to main structure. Flavonoid glycones such as naringin and hesperidin in korean citrus peel are slower absorbed and consequently less active than their aglycone, naringenin and hesperetin, respectively. Therefare to increase the content of flavonoid aglycone in korean citrus peel, we used commercial carbohydrate-hydrolysing enzymes, AMG 300 L, Pectinex 100 L, and Viscozyme for transforming flavonoid glycones to aglycones. Optimal conditions of enzyme reaction were pH 5.0-7.0, $5\%$ enzyme, and 24-48 hrs. The content of naringenin and hesperetin as flavonoid aglycones in untreated citrus peel is $100\~200\;ng/g$ of dried citrus peel. In case of enzyme-treated citrus peel the content of naringenin and hesperetin increased to $1,539\∼6,674\;ng/g\;and\;1,974\∼8,906\;ng/g$ of dried citrus peel, respectively. Finally the content of flavonoid aglycones could be extracted to 10-80 times. Now enzyme-treated citrus peel may be applied to use for functional food because of its higher flavonoid aglycones as more active compounds.