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황토로부터 분리한 Bacillus licheniformis의 항진균 chitinase 생산과 효소 특성
한귀환(Gui Hwan Han),봉기문(Ki Moon Bong),김종민(Jong Min Kim),김평일(Pyoung Il Kim),김시욱(Si Wouk Kim) 한국생물공학회 2014 KSBB Journal Vol.29 No.3
In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod (2~3×0.3~0.4 μm), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLSCU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were 40oC and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.
미생물제(Sphingobium sp. Cam5-1) 처리에 따른 토양 중 카두사포스의 분해효과
연제형 ( Jehyeong Yeon ),정준휘 ( Joon-hui Chung ),최한석 ( Han Suk Choi ),고영준 ( Young-joon Ko ),김다연 ( Dayeon Kim ),안시현 ( Sihyun An ),안재형 ( Jae-hyung Ahn ),한귀환 ( Gui Hwan Han ),원항연 ( Hang-yeon Weon ) 한국환경농학회 2023 한국환경농학회지 Vol.42 No.4
Cadusafos, an organophosphorus insecticide, has been commonly used against various pests worldwide. Organophosphorus pesticides have shorter half-lives and lower toxicities than organochlorine pesticides. However, excessive use of Cadusafos can increase pest resistance and issues with acetylcholine biomagnification, potentially resulting in human toxicity. In this study, we investigated the effect of a Cadusafos-degrading microbial agent (CDMA) prepared using Sphingobium sp. Cam5-1, which was previously reported to effectively degrade residual Cadusafos in soil. Experiments were conducted under both controlled laboratory and greenhouse field conditions. Under laboratory conditions, CDMA (10<sup>6</sup> cfu/g soil application rate) decomposed 97% of Cadusafos in the soil in the untreated control after 21 days. Additionally, when CDMA (10<sup>6</sup> cfu/g soil) was mixed with quicklime, 99% of Cadusafos was decomposed within 3 days. Under greenhouse field conditions, the combined effect of CDMA (10<sup>6</sup> cfu/g soil) and quicklime was not observed. However, CDMA (10<sup>6</sup> cfu/g soil) application alone was capable of decompos- ing 91% of Cadusafos after 3 days. These results indicate that CDMA can effectively decompose high residual levels of Cadusafos in soils under field conditions using a low inoculum rate.
복합기능미생물 Bacillus velezensis GH1-13을 이용한 미생물제형 개발 및 생물학적 특성
박준경(Jun-Kyung Park),서선일(Sun-Il Seo),한귀환(Gui Hwan Han),김공민(Kong-Min Kim),김대혁(Dae-Hyuk Kim),송재경(JaeKyeong Song),김평일(Pyoung Il Kim) 한국생물공학회 2018 KSBB Journal Vol.33 No.4
Biological control of plant diseases including bacterial and fungal pathogens has been deliberated as a viable alternative method to harmful and expensive chemical control. Bacillus velezensis GH1-13 strain was reported as a multifunctional biological agent which inhibits the plant pathogens and utilized in sustainable agriculture for improving growth of the crops. The objective of this study is to evaluate the biological potential of Bacillus velezensis GH1-13 for effective use in agriculture and industrialization. The microbial formulations of GH1-13 strain were performed into liquid, powder, granule and pellet types. Total viable cells of four microbial formulations were verified more than 107 cfu/mL(g) during 8 weeks at 40℃, and the storage stability was considered under severe condition. Plant growth promoting of four formulation types were examined to pepper and tomato plants. The powder type showed the best effect such as stem length 25.6±0.72 cm, 10 petiole and stem length 35.3±0.46 cm, 11 petiole of pepper and tomato plant, respectively. Biocontrol effect of liquid and powder types was also tested against Colletotrichum gloeosporioides causing pepper anthracnose. The resulting, 100 fold diluted liquid type exhibited control effect of 65%, and side effect on pepper and tomato plants were not detected. From the results, we suggest that microbial formulations have significant effects as biological agents such as plant growth promoting and controlling the phytopathogenic fungi. In addition, we expect that microbial agent may be useful in agriculture, although further study is needed.
미생물제(Rhodococcus sp. 3-2) 처리에 따른 토양 중 카벤다짐의 분해효과
연제형 ( Jehyeong Yeon ),김현수 ( Hyeon-su Kim ),안재형 ( Jae-hyung Ahn ),한귀환 ( Gui Hwan Han ),오영곤 ( Young Goun Oh ),조일규 ( Il Kyu Cho ),박인철 ( In-cheol Park ) 한국환경농학회 2021 한국환경농학회지 Vol.40 No.4
BACKGROUND: The fungicide of benomyl, a benzimidazole group, has been commonly used for pesticides against fungal diseases in the world. However, benomyl is rapidly hydrolyzed in the environment after using to control plant diseases and has adverse effects by generating carbendazim, which is toxic to plants, humans, and the environment. METHODS AND RESULTS: In this study, the decomposition effect of carbendazim, a degradation product of benomyl was conducted in pot and field after making a prototype of benomyl-degrading microbial agent (BDMA). We found that the carbendazim-degrading microbial agent (CDMA) (10<sup>5</sup>, 10<sup>6</sup>, and 10<sup>7</sup> cfu/g soil) decomposed carbendazim by 50% or more in all the treatments, compared to the untreated control in the pot tests after four weeks. The effect of 100% decomposition of carbendazim was observed at 7 days after treatment, when the prototype of BDMA was apllied at 10-folds dilution in the field. The decomposition effect at more than 60% and plant growth promoting effect were observed after 7 days of the treatment, compared with the untreated group in the second field experiment,treated with commercially available concentrations of 500-folds and 1,000-folds. CONCLUSION(S): These results might represent that the BDMA would decompose carbendazim effectively, a decomposition product of the fungicide benomyl, remaining in agricultural area, and it could be utilized practically by using a low dilution rate.
카벤다짐 분해 미생물인 Rhodococcus sp. 3-2의 대량 배양 및 미생물 제제 개발
박준경 ( Jun-kyung Park ),임성훈 ( Seonghun Im ),김정원 ( Jeong Won Kim ),지정환 ( Jung-hwan Ji ),김공민 ( Kong-min Kim ),박해성 ( Haeseong Park ),윤영석 ( Yeong-seok Yoon ),원항연 ( Hang-yeon Weon ),한귀환 ( Gui Hwan Han ) 한국환경농학회 2023 한국환경농학회지 Vol.42 No.4
Rhodococcus sp. 3-2 strain has been reported to degrade benzimidazole-based pesticides, such as benomyl and carbendazim. Therefore, this study aimed to optimize culture medium composition and culture conditions to achieve cost-effective and efficient large-scale production of the Rhodococcus sp. 3-2 strain. The study identified that the optimal media composition for mass culture comprised 0.5% glucose, 0.5% yeast extract, 0.15% NaCl, 0.5% K<sub>2</sub>HPO<sub>4</sub>, 0.5% sodium succinate, and 0.1% MgSO<sub>4</sub>. Additionally, a microbial agent was developed using a 1.5-ton fermenter, with skim milk (20%), monosodium glutamate (15%), and vitamin C (2%) as key components. The storage stability of the microbial agent has been confirmed, with advantages of low temperature conservation, which helps to sustain efficacy for at least six months. We also assessed the benomyl degradation activity of the microbial agent within field soil. The results revealed an over 90% degradation rate when the concentration of viable cells exceeded 2.65 × 10<sup>6</sup> CFU/g after a minimum of five weeks had elapsed. Based on these findings, Rhodococcus sp. 3-2 strain can be considered a cost-effective microbial agent with diverse agricultural applications.