• Journal of Microbiology
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• v.45 no.6
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• pp.553-557
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• 2007

In order to detect and identify the most toxic Bacillus thuringiensis strains against pests, we isolated a B. thuringiensis strain (Bn1) from Balaninus nucum (Coleoptera: Curculionidae), the most damaging hazelnut pest. Bn1 was characterized via morphological, biochemical, and molecular techniques. The isolate was serotyped, and the results showed that Bn1 was the B. thuringiensis serovar, kurstaki (H3abc). The scanning electron microscopy indicated that Bn1 has crystals with cubic and bipyramidal shapes. The Polymerase Chain Reactions (PCRs) revealed the presence of the cry1 and cry2 genes. The presence of Cry1 and Cry2 proteins in the Bn1 isolate was confirmed via SDS-PAGE, at approximately 130 kDa and 65 kDa, respectively. The bioassays conducted to determine the insecticidal activity of the Bn1 isolate were conducted with four distinct insects, using spore-crystal mixtures. We noted that Bn1 has higher toxicity as compared with the standard B. thuringiensis subsp. kurstaki (HD-1). The highest observed mortality was 90% against Malacosoma neustria and Lymantria dispar larvae. Our results show that the B. thuringiensis isolate (Bn1) may prove valuable as a significant microbial control agent against lepidopteran pests.

• The Korean Journal of Pesticide Science
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• v.15 no.2
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• pp.166-176
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• 2011

Bacillus thuringiensis CAB530 was isolated from dead Anomata albopilosa (Rutelidae: Coleoptera) and soil of green tea field, and confirmed its insecticidal activities. CAB530 isolate showed a high insecticidal activity against the beet armyworm among the many lepidopteran insects that are difficult to control. $LC_{50}$ value of CAB530 isolate against the second larva of Spodoptera exigua was $1.49{times}10^4$ spore concentration (cfu/$m{\ell}$). SDS-PAGE result of insecticidal toxin protein of CAB530 isolate showed a band at 130 kDa that is similar pattern with B. thuringiensis subsp. kurstaki that took insecticidal activity against S. exigua. Otherwise, the crystal protein of the CAB530 isolate was conformed at 65 kDa level after 30 minute of incubation in S. exigua midgut juice. Six crystal genes (cry1Aa, cry1Ab, cry1C, cry1D, cry1F and cry1I) were identified by PCR. It different from genes of B. thuringiensis subsp. kurstaki. Crystal shape and pattern of toxin protein was similar with B. thuringiensis subsp. kurstaki, however, insecticidal activity and PCR result of CAB530 isolate was similar with B. thuringiensis subsp. aizawai.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.168-173
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• 1988

Immunological analysis between endotoxin proteins produced by Bacillus thuringiensis serovar. kurstaki HD1 and HD73 have been investigated by using polyclonal antibodies. The antisera against the endotoxin proteins were prepared from rabbits injected with the endotoxin protein antigens. When about 2mg/$m\ell$ of the antigens were injected for 7 times, the titers were highest. The stability of the antigens was reduced to about 50% after 9 days incubation at 4$^{\circ}C$. The sensitibity of endotoxin protein from B. thuringiensis HD1 and HD73 by indirect ELISA was 50ng/$m\ell$ and 400ng/$m\ell$, respectively. The cross reaction of antiserum appeared that anti-HD1 partialy reacted with crystal protein but anti-HD73 reacted with HD1 endotoxin about 100%.

• Microbiology and Biotechnology Letters
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• v.14 no.4
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• pp.325-328
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• 1986

The lethality to Pieris raepi larvae and the safety tests to mice of the endotoxin crystals and the pesticide of B thuringiensis serovar kurstaki were carried out. The LD50 of the endotoxin (3.9$\times$10$^9$ spores/$m{\ell}$) appeared to be 1$\mu$g and that of the pesticide (2.6$\times$10$^9$ spores/$m{\ell}$) was about 9 $\mu$g. When the endotoxin solution and the pesticide were injected to the mice's intraperitoneal cavities, 10 to 20% of the mite were dead. Also in the case of intracerebral injection 20% of the mice were dead at the doses of 7.8$\times$10$^3$ spores and 100% of the mice were dead at the concentration of 7.8$\times$10$^8$ spores. but the mice in the oral, subcutaneous and inhalation treatment groups were safe and healthy. When the pesticide applied to the cabbage field for raepi larvae, 52 to 65% of the larvae in the field were killed in 4 days postapplication.

• Journal of Sericultural and Entomological Science
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• v.36 no.2
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• pp.157-161
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• 1994

To investigate the morphology of Bacillus thuringiensis crystal proteins, two type crystal protein genes, cryIA(c) gene under the control of cryIA(b) gene promoter and cryIIA gene under the control of its own promoter, were transformed in B. thuringiensis acrystalliferous mutant strain and the transformants were characterized by SDS-PAGE and scanning electron microscopy. The expression and formation of crystal proteins in B. thuringiensis subsp. kurstaki Cry-B revealed that crystal proteins appear to have same molecular weight and morphology to those of wild type strain's, suggesting that the expression and formation of crystal proteins affected not by host cell or recombination of cryIA(e) gene under the control of cryIA(b) gene promoter but by only structural fragment of protoxin.

• The Korean Journal of Pesticide Science
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• v.14 no.4
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• pp.446-455
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• 2010

The characteristics of parasporal inclusion body from Bacillus thuringiensis subsp. kurstaki KB099 isolate which is high bioactive to the tobacco cutworm, Spodoptera litura, were examined. Parasporal inclusion of B. thuringiensis subsp. kurstaki KB099 isolate showed only 1 band at 130 kDa compared with B. thuringiensis subsp. kurstaki HD-l isolate producing 2 protein bands at 130 kDa and 60 kDa from by SDS-PAGE analysis without any enzyme treatment. Also, we confirmed that gut extract of sensitive S. litura KB099 isolate had digested only 60 kDa ${\delta}$-endotoxin protein. When the digestive enzyme of sensitive insect responsible for parasporal inclusion from KB099 and HD-l isolate was treated to each of them, protein band 60 KDa of KB099 was maintained up to 12 hours but all bands of HD-l were disappeared within 6 hours. In KB099 isolate, 6 genes (Cry1Aa, Cry1Ab, Cry1Ac, Cry1C, Cry1D and Cry1I) were identified by PCR analysis. Also, $Cry^-$ mutant of KB099 isolate was investigated by phase- contrast microscope, SDS-PAGE and PCR.

• Korean journal of applied entomology
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• v.28 no.2
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• pp.69-75
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• 1989

The extracellular amylase production by Bacillus thuringiensis subsp. kurstaki HD-l in amylase production media and its characteristics were investigated. The amylase production was highest in the medium composed of 0.2% soluble starch, 1.0% Bacto-peptone, 0.3% beef extract, 0.3% yeast extract, 0.5% NaCl, 0.3% $K_2HPO_4$, 0.1% $KH_2PO_4$, 0.012% $CaCl_2$.$2H_2O$, 0.005% $MnSO_4$.$H_2O$, and 0.03% $MgSO_4$.$7H_2O$. The amylase activity was inhibited by 50mM EDT A. The enzyme was optimally active from pH 6.5 to 7.0 at $55^{\circ}C$, The specific activity of the enzyme in the ethanol precipitate was 2.01 units/mg, and the Km value was approxi-mately 0.8 mg/ml.

• Journal of Sericultural and Entomological Science
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• v.36 no.2
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• pp.162-167
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• 1994

Surface changes of tissues caused by B. thuringiensis var. kurstaki-$\delta$-endotoxin intoxication of Helicoverpa assulta were observed by scanning electron microscopy. Bt-endotoxin crystals induced the erosion and disruption on the surface of all tissues tested. The results revealed that the toxin binds to all exposed plasma membranes without apparent specificity for particular membrane domains.

• Korean journal of applied entomology
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• v.47 no.2
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• pp.175-184
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• 2008

To screen highly active Bacillus thuringiensis isolates against Spodoptera litura (Lepidoptera, Noctuidae), 46 B. thuringiensis was isolated from 115 samples obtained from several crop soils. Especially, B. thuringiensis subsp. kurstaki CAB133 and CAB162 isolates showed 100% mortality against S. litura. $LD_{50}$ values of CAB 133, CAB162 and HD-1 strains of B. thuringiensis subsp. kurstaki were 0.089, 3.144 and $0.513{\mu}g/ml$ against 2nd larva of S. litura, respectively. The weight of 3rd larva of S. litura which were fed crystal inclusion protein $(1.267{\mu}g/ml)$ with B. thuringiensis subsp. kurstaki CAB133 was about 30 times lass than control group. CAB133 and CAB 162 strains of B. thuringiensis subsp. kurstaki which were taken a highly toxity against S. litura were analyzed by SDS-PAGE, and estimated the molecular weight of the Cry proteins. Their serological identification by H serotypes were showed B. thuringiensis subsp. kurstaki (3abc) type.

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1836-1840
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• 2008

Bacteriocins are antimicrobial peptides that are produced by bacteria and toxic to bacterial strains closely related to the producer strain. It has previously been reported that Bacillus thuringiensis strain NEB17 and Bacillus thuringiensis subsp. kurstaki BUPM4 produce the bacteriocins thuricin 17 (3,162 Da) and bacthuricin F4 (3,160.05 Da), respectively. Here, we demonstrate that these bacteriocins have functional similarities and show a similar spectrum of antimicrobial activities against indicator strains. We also studied the effects of sterilization methods on the recovery and biological activities of these bacteriocins. They were completely degraded by autoclaving and the two were similarly affected by the tested filter membranes. Polyvinylidene fluoride (PVDF), polyestersulfone (PES), and cellulose acetate (CA) are suitable for filter sterilization of these bacteriocins. The two bacteriocins were stable across a range of storage conditions. These data will facilitate their utilization in food preservation or agricultural applications.