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Polyvinyl Alcohol 이용 공생균 Pseudomonas J2W 균주와 Xanthomonas J2Y 균주의 배양조건
조윤래 영남대학교 자원문제연구소 1992 資源問題硏究 Vol.11 No.-
In preceding studies, two strains Pseudomonos J2W and Xanthomonos J2Y which can utilize polyvinyl alcohol (PVA) as a sole carbon source were isolated from soil. PVA was utilized symbiotically by mixed culture of these two strains. The optimum culture conditions for the symbiotic PVA utilization of the strains were investigated. The optimum temperature and pH for growth of the mixed culture were 28-30℃ and 7.0, respectively. And then, when the agitation speed of the mixed culture of symbionts was increased, the degradation rate of PVA of theirs was increased. Ammonium sulfate, urea and peptone. were effective as nitrogen sources. and then, vitamin BH was appeared to have a significant influence on growth of the mixed culture of symbionts. When the symbionts were cultivated in the rich medium, the degradation rate of PVA of theirs was significantly decreased.
Bacillus sp. SSA3 균주의 Expression Vector 開發
조윤래,김종규,권대준 한국산업미생물학회 1992 한국미생물·생명공학회지 Vol.20 No.6
한국 재래식 된장·간장 발효균 Bacillus sp. SSA3 균주의 expression vector를 개발하기 위해 Bacillus sp. SSA3의 chromosomal DNA로부터 유전자의 promoter 부위를 cloning하였다. Recombinant plasmid를 제작하기 위해 Bacillus sp. SSA3의 chromosomal DNA을 HindⅢ로 절단한 단편을 pGR71 plasmid의 CAT gene과 pUC18 plasmid의 β-galactosidase gene의 전방에 삽입시킨 후, E. coli JM109에 형질 전환하였다. E. coli JM109의 chloramphenicol 내성 clones으로부터 6 recombinant plasmid를 선별하였다. 이들 선별된 plasmid는 Bacillus sp. SSA3와 E. coli JM109에서 promoter의 활성이 확인되었다. 그리고 실제 간장·된장 발효시 여러 농도의 염이 첨가되므로 이 선별된 재조합 plasmid를 Bacillus sp. SSA3의 expression vector로 사용시 각 재조합 plasmid 중에 삽입된 promoter의 염에 대한 영향의 정도를 확인하기 위해 10% NaCl이 첨가된 LB medium상에서 배양하였을 때, 이들 중 Bacillus sp. SSA3의 4 clones은 융합 CAT gene의 발현이 강하게 감소되었으나 2 clones은 약하게 저해되었다. The promoter regions from chromosomal DNA of Bacillus sp. SSA3 which is responsible for fermentation of Korean traditional soy sauce, were cloned for construction of expression vector of Bacillus sp. SSA3. Recombinant plasmids were constructed by insertion of HindⅢ-cleaved Bacillus sp. SSA3 chromosomal DNA fragments in front of the CAT gene of pGR71 plasmid and β-galactosidase gene of pUC18 plasmid. 6 recombinant plasmids were isolated from chloramphenicol resistant E. coli JM109 clones. All these plasmids were found to have promoter activity in Bacills sp. SSA3 and E. coli JM109. When these 6 clones of Bacills sp. SSA3 were cultivated in LB agar medium supplemented with 10% NaCl, fused CAT gene expression of 4 clones was significantly decreased in common. But the others were poorly inhibited.
Polyvinyl alcohol 이용 공생균 Pseudomonas sp . J2W 와 Xanthomonas sp . J2Y 의 특성
조윤래 한국농화학회 1992 Applied Biological Chemistry (Appl Biol Chem) Vol.35 No.1
Two strains J2W and J2W which were isolated from soil can utilize polyvinyl alcohol(PVA) as a sole carbon source. PVA was utilized symbiotically by the mixed culture of these two strains which could not utilize PVA in each respective pure culture. Effect of degree of PVA polymerization on the its utilization was examed, and there was remarkable difference among three kind of PVA(PVA 500, 1500 and 2000). The reconstruction of there two strains was carried out with other symbionts Pseudomonas sp. PW and Pseudomonas sp. G5Y which were able to utilize PVA. PVA utilization occured in each remixed culture of J2Y strain with Pseudomonas sp. PW J2W strain with Pseudomonas sp. G5Y, respectively. Identification of bacteria was based on morphological and biological chatacteristics, J2W and J2Y strain were similar to a strain of Pseavdomonas Pseudimallei and Xanthomonas campestris, respectively.