• Title/Summary/Keyword: High temperature incubation

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Modification of the commercial silkworm eggs adequate for Bluemoon0silkworm transgenesis (누에 형질전환에 적합한 실용품종 누에알의 제조)

  • Kim, Sung-Wan;Kang, Min-Uk;Kang, Seok-Woo;Yun, Eun-Young;Choi, Kwang-Ho;Kim, Seong-Ryul;Park, Seung-Won;Nho, SiKab;Goo, Tae-Won
    • Journal of Sericultural and Entomological Science
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    • v.51 no.1
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    • pp.73-77
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    • 2013
  • Silkworm transgenesis scientists have done some genetic modification work on multivoltine silkworms, but that type of silkworms is less commercial feasible. They are easy to manipulate, because they breed all year round. But the commercial silkworm variety must undergo hydrochloric acid treatment at a high temperature to be artificially hatched. Hydrochloric acid penetrates through the holes in the silkworm eggs, fatally damaging their reproduction. So it had been thought that altering the properties of the commercial silkworm variety would be very difficult. So we have tried to make from diapause to non-diapause eggs using diapauses varieties, 'Backokjam' and 'Jam 124'. At present, our group has establishing the conditions for non-diapause eggs. Oviposited eggs after 40 ~ 60 hours were incubated for 24 hours at $15{\sim}20^{\circ}C$ with dark condition. Non-diapause eggs were completely induced. The hatching rate, molting rate and pupation rate of non-diapause 'Jam 124' and 'Backokjam' eggs showed no differences compared to diapause eggs. When transgenic silkworm using the non-diapause eggs, the hatching rate showed that non-diapause eggs induced from diapause were 40 ~ 70%, diapause eggs treated with artificial incubation were 10 ~ 30%, and polyvoltine strains, HM eggs were 30 ~ 50%. Therefore, we suggest that modification techniques of the commercial silkworm eggs adequate for silkworm transgenesis can be used to develop transgenic silkworms more easily.

Characterization of Phenanthrene Degradation by Sphingomonas sp. HS362 (Sphingomonas sp. HS362에 의한 Phenanthrene 분해특성)

  • Kim Su Hwa;Hong Seung-Bok;Kang Hee Jeong;Ahn Jin-Chul;Jeong Jae Hoon;Son Seung-Yeol
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.201-207
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    • 2005
  • A phenanthrene-degrading bacterium HS362, which is capable of using phenanthrene as a sole carbon and energy source, was isolated from oil contaminated soil. This strain is a gram negative, rod shaped organism that is most closely related to Sphingomonas paucimobilis based on biochemical tests, and belongs to the genus Sphingomonas based on fatty acids analysis. It exhibited more than $99.2{\%}$ nucleotide sequence similarity of 16S rDNA to that of Sphingomonas CF06. Thus, we named this strain as Sphingomonas sp. HS362. It degraded $98{\%}$ of phenanthrene after 10 days of incubation when phenanthrene was added at 500 ppm and $30{\%}$ even when phenanthrene was added at 3000 ppm. Sphingomonas sp. HS362 could also degrade low molecular weight PAHs(Polycyclic aromatic hydrocarbons) such as indole and naphthalene, but was unable to degrade high molecular weight PAHs such as pyrene and fluoranthene. The optimum temperature and pH for phenanthrene degradation were $30^{\circ}C$ and $4{\~}8$, respectively. Sphingomonas sp. HS362 could degrade phenanthrene effectively in the concentration range of NaCl of up to $1{\%}$. Its phenanhrene degrading ability was enhanced by preculture, suggesting the possibility of induction of phenanthrene degrading enzymes. Starch and surfactants such as SDS, Tween 85, and Triton X-100 were also able to enhance phenanthrene degradation by Sphingomonas sp. HS362. It carries five plasmids and one of them, plasmid p4, is considered to be involved in the degradation of phenanthrene according to the plasmid curing experiment by growing at $42^{\circ}C$.

Inhibition of Listeria monocytogenes by Low Concentrations of Ethanol (저농도의 Ethanol에 의한 Listeria monocytogenes의 증식억제)

  • 박찬성;김미림
    • Korean journal of food and cookery science
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    • v.11 no.4
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    • pp.379-385
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    • 1995
  • The effect of low concentrations of ethanol (3-7%, v/v) in tryptic soy broth (TSB) as an antibacterial agent against Listeria monocytogenes was tested at -20, 5, 35, 45, 50 and 55$^{\circ}C$. Increasing concentrations of ethanol progressively inhibited initial growth of L. monocytogenes at 35$^{\circ}C$. Growth occured at 5% ethanol, but only after a prolonged lag period. The number of viable cells of L. monocytogenes declined during incubation at 7% ethanol. TSB containing 3-7% ethanol was inoculated with 10$\^$5/-10$\^$6/ cells/$m\ell$ or L. monocytogenes and incubated at low temperatures (5$^{\circ}C$, -20$^{\circ}C$). In the presence of 3% of ethanol at 5 or -20$^{\circ}C$, bacterial growth was inhibited more than 90% of control cells. TSB containing 3-7% ethanol was inoculated with 10$\^$6/-10$\^$7/ cells/$m\ell$ of L. monocytogenes and incubated at high temperatures (45$^{\circ}C$, 50$^{\circ}C$, 55$^{\circ}C$). Decrease in viability of the cells incubated at 45 or 50$^{\circ}C$ was slow and the survival of L. monocytogenes was not affected so much in the presence of 3% of ethanol. The viability of L. monocytogenes was decreased with increasing concentration of ethanol and temperature. Decimal reduction times (D-values) based on tryptic soy agar plates at 55$^{\circ}C$ were 20.1, 12.6, 7.4 and 4.2 min in 0, 3, 5 and 7% ethanol, respectively.

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Psychrophilic oyster mushroom variety 'Seolan' (느타리버섯 신품종 육성 연구 -저온성 느타리버섯 신품종 '설안'-)

  • Chi, Jeong-Hyun;Choi, Jong-In;Ha, Tai-Moon;Ju, Young-Cheul
    • Journal of Mushroom
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    • v.4 no.3
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    • pp.116-121
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    • 2006
  • 'Seoln', a new variety of oyster mushroom was developed by Mushroom Research Institute, Gyonggi Province Agricultural Research and Extension Services in 2005. It was bred with mating between monokaryotic strains isolated from KME35124 and $F{\cdot}X3$. The major characteristics of the mushroom was shown a lot of primordia, the gray-colored and bunch pileus. The optimum temperature for the mycelial growth was around $26{\sim}30^{\circ}C$ and that for the primordia and growth of fruit-body was around $10{\sim}15^{\circ}C$. Incubation period was required around 25 days with bottle culture and about 23 days in Poly Prophylene(P.P) plastic bag culture. The yield was shown high by 138.8g/bottle and 250.9%/P.P bag.

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Expression and Characterization of Calcium- and Zinc-Tolerant Xylose Isomerase from Anoxybacillus kamchatkensis G10

  • Park, Yeong-Jun;Jung, Byung Kwon;Hong, Sung-Jun;Park, Gun-Seok;Ibal, Jerald Conrad;Pham, Huy Quang;Shin, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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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.

Ruminal Behavior of Protein and Starch Free Organic Matter of Lupinus Albus and Vicia Faba in Dairy Cows

  • Yu, P.;Leury, B.J.;Egan, A.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.974-981
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    • 2002
  • Faba beans (vicia faba) (FB) and lupin seeds (Lupinus Albus) (LS) were dry roasted at three temperatures (110, 130, $150^{\circ}C$) for 15, 30 or 45 min to determine the effects of dry roasting on rumen degradation of crude protein and starch free organic matter ($^{PSF}OM$). Rumen degradation characteristics of $^{PSF}OM$ were determined by the nylon bag incubation technique in dairy cows fed 60% hay and 40% concentrate. Measured characteristics of $^{PSF}OM$ were undegradable fraction (U), degradable fraction (D), soluble fraction (S), lag time (T0), and the rate of degradation (Kd). Based on the measured characteristics, rumen availability ($RA^{PSF}OM$) and bypass $^{PSF}OM$ ($B^{PSF}OM$) were calculated. Dry roasting did not have a greater impact on rumen degradation characteristics of $^{PSF}OM$ (p>0.05). S varied from 32.1 (raw) to 30.0, 27.8, 30.8% (LS) and 15.4 (raw) to 14.4, 20.8, 20.9% (FB); D varied from 65.4 (raw) to 66.3, 66.9, 55.9% (LS) and 54.9 (raw) to 55.0, 51.0, 64.7% (FB); U varied from 2.6 (raw) to 7.3, 7.0, 7.7% (LS) and 29.7 (raw) to 30.6, 28.2, 14.4% (FB); Kd varied from 6.0 (raw) to 7.3, 7.0, 7.7% (LS) and 22.4 (raw) to 24.4, 21.1, 7.9% (FB); $B^{PSF}OM$ varied from 35.5 (raw) to 33.8, 36.6, 38.2% (LS) and 41.3 (raw) to 41.5, 39.7, 47.6% (FB) at 110, 130 and $150^{\circ}C$, respectively. Therefore dry roasting did not significantly affect $RA^{PSF}OM$, which were 353.7, 367.9, 349.6, 336.9 (g/kg DM) (LS) and 12.82, 127.0, 133.7, 117.1 (g/kg DM) (FB) at 110, 130 and $150^{\circ}C$, respectively. These results alone with our previously published reports indicate dry roasting had the differently affected pattern of rumen degradation characteristics of various components in LS and FB. It strongly increased bypass crude protein (BCP) and moderately increased starch (BST) with increasing temperature and time but least affected $^{PSF}OM$. Such desirable degradation patterns in dry roasted LS and FB might be beneficial to the high yielding cows which could use more dry roasted $^{PSF}OM$ as an energy source for microbial protein synthesized in the rumen and absorb more amino acids and glucose in the small intestine.

Immobilization of Cellulases from Fomitopsis pinicola and Their Changes of Enzymatic Characteristics (흡착법에 의한 Fomitopsis pinicola 유래 cellulase의 고정화와 그에 따른 효소특성 변화)

  • Shin, Keum;Kim, Tae-Jong;Kim, Young-Kyoon;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.3
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    • pp.251-261
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    • 2010
  • Cellulase from Formiptosis pinicola KMJ812 is an efficient cellulose degradation enzyme complex, especially with a high ${\beta}$-glucosidase activity. In this study, the change in enzymatic characteristics by immobilization and the reduction of immobilized enzyme activity by repeated usages were evaluated using cellulases from F. pinicola KMJ812. Among tested four resins, Duolite A568 resin had the best enzyme activity yield with 61.7% cellulase activity and 64.4% ${\beta}$- glucosidase activity during the cellulase immobilization. The best reaction temperature was $55^{\circ}C$ for both cellulase and ${\beta}$-glucosidase activities which were higher than the unimmobilized soluble cellulases. The best reaction pH was 4.0 for cellulase activity which was a little more basic than a soluble form and 4.5 for ${\beta}$-glucosidase activity. The immobilized cellulase activity was remained 98% of the beginning activity after 72 h incubation at $50^{\circ}C$ and 50% of the beginning activity after eight times usage at $50^{\circ}C$.

Characteristics of a new oyster mushroom variety 『Dajoa』 for the bottle and poly prophylene plastic bag culture (느타리버섯의 신품종 육성 연구 -병·봉지재배용 신품종 『다조아』 느타리버섯의 특성-)

  • Chi, Jeong-Hyun;Choi, Jong-In;Ju, Young-Cheul
    • Journal of Mushroom
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    • v.3 no.2
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    • pp.60-64
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    • 2005
  • 'Dajoa', a new variety of oyster mushroom was developed by Mushroom Research Institute, Gyonggi Province Agricultural Research and Extension Services in 2004. It was bred with mating between monokaryotic strains isolated from Boonli 89-1 and ASI 2018-249. The major characteristics of the mushroom are showing a lot of pinheadings, the gray-colored and infundibuliform pileus. The optimum temperature for the mycelial growth was around $26{\sim}28^{\circ}C$ and that for the pinheading and growth of fruitbody was around $15{\sim}18^{\circ}C$. Incubation period was required around 24 days with bottle culture and about 21days in Poly Prophylene(P.P) plastic bag culture. The yields were shown high by 140.7g/850cc bottle and 260.3g/1kg P.P. plastic bag.

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Isolation and characterization of a novel gossypol-degrading bacteria Bacillus subtilis strain Rumen Bacillus Subtilis

  • Zhang, Yunhua;Zhang, Zhengyou;Dai, Li;Liu, Ying;Cheng, Maoji;Chen, Lijuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.63-70
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    • 2018
  • Objective: The aim of the study was to isolate gossypol-degrading bacteria and to assess its potential for gossypol degradation. Methods: Rumen liquid was collected from fistulated cows grazing the experimental pasture. Approximately 1 mL of the rumen liquid was spread onto basal medium plates containing 2 g/L gossypol as the only source of carbon and was then cultured at $39^{\circ}C$ to isolate gossypol-degrading bacteria. The isolated colonies were cultured for 6 h and then their size and shape observed by microscope and scanning electron microscope. The 16S rRNA gene of isolated colonies was sequenced and aligned using National Center for Biotechnology Information-Basic Local Alignment Search Tool. The various fermentation conditions, initial pH, incubation temperature, inoculum level and fermentationperiod were analyzed in cottonseed meal (CSM). The crude protein (CP), total gossypol (TG), and free gossypol (FG) were determined in CSM after fermentation with isolated strain at $39^{\circ}C$ for 72 h. Results: Screening results showed that a single bacterial isolate, named Rumen Bacillus Subtilis (RBS), could use gossypol as a carbon source. The bacterium was identified by 16S rDNA sequencing as being 98% homologous to the sequence of Bacillus subtilis strain GH38. The optimum fermentation conditions were found to be 72 h, $39^{\circ}C$, pH 6.5, moisture 50%, inoculum level $10^7cell/g$. In the optimum fermentation conditions, the FG and TG content in fermented CSM decreased 78.86% and 49% relative to the control. The content of CP and the essential amino acids of the fermented CSM increased respectively, compared with the control. Conclusion: The isolation of a gossypol-degrading bacterium from the cow rumen is of great importance for gossypol biodegradation and may be a valuable potential source for gossypol-degradation of CSM.

Investigation of Fungicides Inhibitory Effect of on Summer Patch Disease, Caused by Magnaporthiopsis poae, in Kentucky bluegrass (여름잎마름병(Summer patch) 병원균에 대한 살균제의 억제효과 조사)

  • Lee, Jung Han;Shim, Gyu Yul;Kim, Jeong Ho;Jeon, Chang Wook;Kwak, Youn-Sig
    • Weed & Turfgrass Science
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    • v.6 no.2
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    • pp.151-156
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    • 2017
  • Summer patch is the most serious disease at turfgrass field or golf course established with Kentucky bluegrass during high temperature season in Korea. Nevertheless, chemicals for the summer patch control are not yet registered in Korea. We isolated the pathogens from the turfgrass showing typical summer patch symptoms and identified as Magnaporthiopsis poae by using the internal transcribed spacer ITS1 and ITS4 sequences of rDNA. The inhibition rates of the pathogen were investigated for 10 fungicides. As results, the pathogen growth was suppressed when chemicals concentration increased and negatively correlated with incubation period with the chemicals. In triazole group, all chemicals (metconazole, myclobutanil, propiconazole and tebuconazole) treated showed the inhibition rates by 100%. Thiophanate-methyl showed the next highest inhibition effect against a summer patch pathogen. In strobilurin group, pyraclostrobin was the highest suppression effect compared with azoxystrobin and trifloxystrobin. Inhibition effect of fludioxonil and fluxapyroxad on pathogen was similar to the trifloxystrobin. Based on the results, triazole and carboxamide groups are strongly recommended due to the highest inhibition effect on the summer patch pathogen, Magnaporthiopsis poae.