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Bacillus thuringiensis 생장과 살충성 결정단백질 생성에 대한 탄소원의 영향
김무기,안병구,Kim, Moo-Key,Ahn, Byung-Koo 한국응용생명화학회 1996 Applied Biological Chemistry (Appl Biol Chem) Vol.44 No.1
Effects of 14 carbohydrates supplied as carbon sources on cell growth and sporulation of, and the production of insecticidal crystal proteins by Bacillus thuringiensis strains were investigated in liquid cultures. Strains grew well in media containing any one of the 14 carbohydrates supplied, reaching maximum cell densities of $10^7{\sim}10^8\;cells/ml$ in 16.7 to 22 hours after inoculation depending on the strain. Spores first appeared in 16.7 to 24.7 hours after inoculation, and 80% sporulation was reached in 28 to 51.3 hours after inoculation depending on the strain. No change in pH of media was observed after cell multiplication. The production of total protein was highest when supplied with sucrose and was lowest with starch. More insecticidal crystal proteins were produced when supplied with glucose, lactose, maltose, or sucrose. The amount of insecticidal crystal proteins produced by the strains was proportional to that of the total protein. The relative amount of individual insecticidal crystal protein species produced by B.t. kurstaki and B.t. israelensis was not influenced by the carbohydrates supplied. 액체배양 실험으로 14가지 탄수화물을 사용하여 B. thuringiensis의 생장, 포자형성 및 살충성 결정단백질 생성에 대한 탄소원의 영향을 조사하였다. 최대 세포밀도는 B. thuringiensis 균주에 따라 접종 $16.7{\sim}22$시간 후에 모든 탄소원배지에서 $10^7{\sim}10^8\;cells/ml$ 수준으로 나타났고, 접종 $16.7{\sim}24.7$시간 후에 포자가 나타나기 시작하여 포자형성율이 80%에 이르는 시간은 균주에 따라 $28{\sim}51.3$ 시간이 소요되었다. 배양에 따른 배지의 pH변화는 없었고, 단백질 총량은 sucrose를 사용한 배지에서 가장 높았고, 전분을 첨가했을때 가장 낮았다. Glucose, lactose, maltose 또는 sucorse를 탄소원으로 사용한 배지에서 살충성 결정단백질 생성량이 많았고, 단백질 총량과 살충성 결정단백질량은 비례관계에 있었다. B.t. kurstaki와 B.t. israelensis에서 생성되는 서로 다른 종류의 살충성 결정 단백질의 양은 사용한 모든 탄소원의 경우 그 개별적 증감의 경향이 같았다.
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Bacillus thuringiensis 생장과 살충성 결정단백질 생성에 대한 탄소원의 영향
김무기(Moo Key Kim),안병구(Byung Koo Ahn) 한국응용생명화학회 1996 Applied Biological Chemistry (Appl Biol Chem) Vol.39 No.3
Effects of 14 carbohydrates supplied as carbon sources on cell growth and sporulation of, and the production of insecticidal crystal proteins by Bacillus thuringiensis strains were investigated in liquid cultures. Strains grew well in media containing any one of the 14 carbohydrates supplied, reaching maximum cell densities of 10^7∼10^8 cells/㎖ in 16.7 to 22 hours after inoculation depending on the strain. Spores first appeared in 16.7 to 24.7 hours after inoculation, and 80% sporulation was reached in 28 to 51.3 hours after inoculation depending on the strain. No change in pH of media was observed after cell multiplication. The production of total protein was highest when supplied with sucrose and was lowest with starch. More insecticidal crystal proteins were produced when supplied with glucose, lactose, maltose, or sucrose. The amount of insecticidal crystal proteins produced by the strains was proportional to that of the total protein. The relative amount of individual insecticidal crystal protein species produced by B.t. kurstaki and B.t. israelensis was not influenced by the carbohydrates supplied.
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Bacillus thuringiensis 의 CryI 과 CytA 단백질의 형태와 용해도에 대한 숙주의 영향
안병구,김무기 ( Byung Koo Ahn,Moo Key Kim ) 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.41 No.1
The cryIB, truncated cryIB[cryIB(α)], cryIA(b), and cytA genes, encoding 135-, 89-, 131-, and 27-kDa proteins, respectively, from Bacillus thuringiensis were cloned into a shuttle vector pBES and expressed in E. coli and Bacillus species. The morphology and solubility in alkaline buffer of the insecticidal crystal proteins were investigated. Transformation of intact cells of E. coli and Bacillus species was achieved by electroporation. High field strength of 11.0 kV/㎝ and resistance of 129 ohms were required for efficient transformation of E. coli strains and 4.5 kV/㎝ and 48 ohms for Bacillus species. Strains of recombinant E. coli and Bacillus species produced the insecticidal crystal proteins and accumulated as the same bipyramidal and irregular structures as those of CryIB and IA(b) and CytA of B. thuringiensls, respectively. The insecticidal crystal proteins accumulated in recombinant E. coli wire smaller in size than those in recombinant Bacillus species. The solubility in alkaline buffer of the insecticidal crystal proteins of recombinant E. coli increased gradually as the pH increased, whereas in the case of Bacillus species the solubility increased gradually as the pH increased up to 9 and then the solubility increased greatly up to two times higher than that of E. coli proteins.
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Bacillus thuringiensis 의 살충성 결정단백질 생성에 대한 질소원의 영향
안병구,김무기 전북대학교 농업과학기술연구소 1996 農大論文集 Vol.27 No.1
Effects of various nitrogenous materials supplied as nitrogen sources on cell growth and sporulation of, and on the production of insecticidal crystal proteins by Bacillus thuringiensis strains were investigated in liquid cultures. Strains grew well in media containing any one of the nitrogenous materials supplied except ammonium sulfate, reaching maximum cell densities of 107~l08 cells/mL in 14.7 to 20.7 hours after inoculation depending on the strain. The growth of the strains, however, was much limited in medium containing ammonium sulafte, reaching maximum cell densities of 105~ 106 cells/mL. Cell growth was extremely slow in medium containing cotton seed hydrolysate: maximum cell density, however, was very high with the highest spore density. Spores first appeared in 18 hours after inoculation, and a minimum of 70% sporulation was reached in 34 hours after inoculation depending on the strain. Production of both total and individual insecticidal crystal proteins was highest when supplied with casamino acid. The amount of insecticidal crystal proteins produced by the strains was proportional to total protein contents and rate of sporulation. The total amount of different insecticidal crystal proteins produced by .B.t. kurstaki and B.t.kurstaki and B.t. israelensis was also proportional to that of the total protein, and the relative amount of individual insecticidal crystal protein species was not influenced by the nitrogenous materials supplied.
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