• Journal of Applied Biological Chemistry
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• 제59권2호
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• pp.165-169
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• 2016

The quantitative analytical method for the bioactive substance, 3-cyano-4-methoxy-N-methyl-2-pyridone (ricinine) and an index compound, ricinoleic acid in castor plant (Ricinus communis) extract or oil was developed. For the determination of a pyridone alkaloid compound, ricinine, successive cartridge cleanup method combined with ultra-performance liquid chromatography was set up with $ENVI-Carb^{TM}$ (0.5 g) and $C_{18}$ SPE cartridges. Accuracy and precision were evaluated through fortification studies of one biopesticide (PE) at 10 and $100mg\;kg^{-1}$. Mean recoveries of ricinine were 98.7 and 96.0 % associated with less than 10 % RSD, respectively. For the determination of ricinoleic acid in castor extract and oil, saponification and methylation were optimized using gas chromatography-time of flight mass spectrometry. Recovery was more than 84.8 % associated with 6.2 % RSD after derivatization procedure. Both methodologies developed were applied to analyze real samples including three castor oil products and six commercially available biopesticides containing R. communis, collected at Korean market. The contents of ricinine and ricinoleic acid in most commercial biopesticides were less than the oil or extract contents indicated by label.

• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.452-458
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• 2015

The biocatalytic efficiency of recombinant Corynebacterium glutamicum ATCC 13032 expressing the secondary alcohol dehydrogenase of Micrococcus luteus NCTC2665 was studied. Recombinant C. glutamicum converts ricinoleic acid to a product, identified by gas chromatography/mass spectrometry as 12-ketooleic acid (12-oxo-cis-9-octadecenoic acid). The effects of pH, reaction temperature, and non-ionic detergent on recombinant C. glutamiucm whole cell bioconversion were examined. The determined optimal conditions for production of 12-ketooleic acid are pH 8.0, 35℃, and 0.05 g/l Tween80. Under these conditions, recombinant C. glutamicum produces 3.3 mM 12-ketooleic acid, with a 72% (mol/mol) maximum conversion yield, and 1.1 g/l/h volumetric productivity in 2 h; and 3.9 mM 12-ketooleic acid, with a 74% (mol/mol) maximum conversion yield, and 0.69 g/l/h maximum volumetric productivity in 4 h of fermentation. This study constitutes the first report of significant production of 12-ketooleic acid using a recombinant Corynebacterium glutamicum-based biocatalyst.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.621-634
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• 2001

피마자유와 메탄올의 transesterification 반응 (methanolysis)에서 Novozym 435가 가장 좋은 활성을 나타냈다. Novozym 435의 경우 유기용매를 사용하지 않을 경우에도 좋은 활성을 유지하였고 사용되는 효소량에 따라 최적의 유기용매가 다르다는 것을 확인하였다. 0.5 wt% Novozym 435를 사용한 경우 DIPE를 사용하였을 매 가장 높은 활성을 나타내었으나, 1.0 wt% 의 경우에는 heptane을 이용하였을 때 가장 높은 활성을 보였다. Novozym 435를 이용한 피마자유의 methanolysis에서 methanol 에 의한 저해 반응이 나타나고 oil과 methanol의 몰 비 가 약 1:7.37 일 경우 최대의 ricinoJeic acid methyl ester 를 얻을 수 있었다. 1.0wt%의 Novozym 435를 이용한 methanolysis를 통하여 하루동안 피마자유로부터 ricinoleic acid methyl ester 를 86.1% 의 수율로 얻을 수 있었다.

• 한국약용작물학회지
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• 제16권5호
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• pp.301-305
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• 2008

수집종 아주까리 40종의 유전자원을 농촌진흥청 농업생명공학연구원 유전자원과로부터 분양받아 바이오디젤 가능 작물로서 조지방 및 지방산 분석결과는 다음과 같다. 수집종 아주까리 종자의 gas chromatogram은 6개의 피크를 보였고, ricinoleic acid의 retention time은 12.7분대이었다. 아주까리 수집종의 평균 기름 함량은 $44.6{\sim}49.4%$이었으며, 최저 41.4%와 최고 52.2%로 10.8%의 차이가 있었다. 지방산 조성은 불포화지방산이 97.6%로 대부분이었으며, 포화지방산은 2.4% 정도로 낮은 함량을 보였다. 지방산 조성은 87.3%가 ricinoleic acid이었으며, oleic acid와 linoleic acid가 각각 4.6%, 5.2%이었다. palmitic acid와 stearic acid의 함량은 1% 내외로 차이가 미미하였고, linolenic acid의 함량은 0.6% 정도로 가장 낮았다.

• Journal of Applied Biological Chemistry
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• 제49권4호
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• pp.131-134
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• 2006

The in vitro anti-fungal activity of hydroxylated fatty acids obtained from microbial conversion by Psuedomonas aeruginosa PR3 using ricinoleic acid(RA), eicosadienoic acid(EDA) and conjugated linoleic acid(CLA) as substrates, was investigated. Bioconverted hydroxylated fatty acids showed different anti-fungal activities potentials against the range of phytopathogenic fungi such as Botrytis cinerea, Rhizoctonia solani, Fusarium oxysporum, Sclerotonia sclerotiorum, Colletotricum capsici, Fusarium solani and Phytophthora capsici. RA and EDA showed up to 50% fungal mycelial inhibition at the concentration of $5{\mu}l\;ml^{-1}$. RA, EDA and CLA also exhibited anti-fungal activities with minimum inhibitory concentration(MIC), ranging from 500 to $1000{\mu}g\;ml^{-1}$. Screening was also carried out using varied concentrations of bioconverted RA and EDA for determining the anti-fungal effect on the spore germination of different fungi. Bioconverted RA and EDA showed a considerable degree of spore germination inhibition.

• Journal of Microbiology and Biotechnology
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• 제29권7호
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• pp.1071-1077
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• 2019

Natural gamma-decalactone (GDL) produced by biotransformation is an essential food additive with a peach-like aroma. However, the difficulty of effectively controlling the concentration of the substrate ricinoleic acid (RA) in water limits the biotransformation productivity, which is a bottleneck for industrialization. In this study, expanded vermiculite (E-V) was utilized as a carrier of RA to increase its distribution in the medium. E-V and three commonly used organic compounds were compared with respect to their effects on the biotransformation process, and the mechanism was revealed. Scanning electron microscopy, Fourier transform infrared spectroscopy and thermogravimetric analysis indicated that RA was physically adsorbed onto the surface of and inside E-V instead of undergoing a chemical reaction, which increased the opportunity for interactions between microorganisms and the substrate. The highest concentration of GDL obtained in the medium with E-V was 6.2 g/l, which was 50% higher than that in the reference sample. In addition, the presence of E-V had no negative effect on the viability of the microorganisms. This study provides a new method for producing natural GDL through biotransformation on an industrial scale.

• Journal of Microbiology and Biotechnology
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• 제25권5호
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• pp.704-708
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• 2015

For the analysis of multiple-reaction intermediates for long-chain dicarboxylic acid biotransformation, simple and reproducible methods of extraction and derivatization were developed on the basis of gas chromatography with flame ionization detector (GC-FID) instead of mass spectrometry. In the derivatization step, change of the ratio of pyridine to MSTFA from 1:3 to 9:1 resulted in higher peak intensity (p = 0.021) and reproducibility (0.6%CV) when analyzing 32 g/l ricinoleic acid (RA). Extraction of RA and ω-hydroxyundec-9-enoic acid with water containing 100 mM Tween 80 showed 90.4-99.9% relative extraction efficiency and 2-7%CV compared with those with hydrophobic ethyl acetate. In conclusion, reduction of the pyridine content and change of the extraction solvent to water with Tween 80 provided compatible derivatization and extraction methods to GC-FID-based analysis of longchain carboxylic acids.

• 한국환경농학회지
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• 제36권1호
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• pp.17-21
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• 2017

BACKGROUND: Castor oil and wormseed extract are important active ingredients for biopesticide, and ricinoleic acid in castor oil and three monoterpenes (ascaridole, carvacrol and p-cymene) in wormseed extract are known bioactive substances. However, their stabilities had not been studied, even though the stability was the core property for estimation of shelf-life of biopesticide. Aimed to investigate the thermal stabilities of the bioactive substances in castor oil and wormseed extracts. METHODS AND RESULTS: The contents of ricinoleic acid and three monoterpenes (ascaridole, carvacrol and p-cymene) were analyzed by gas chromatography (GC). The thermal stabilities of the bioactive substance were measured at $0^{\circ}C$, $23^{\circ}C$, $30^{\circ}C$, $40^{\circ}C$, $45^{\circ}C$ and $54^{\circ}C$ for 84 d. The half-lives of ricinoleic acid in biopesticides was ranged from 28.9 d to 57.8 d at $30^{\circ}C$, and the stability of pure castor oil were located in the range ($t_{1/2}$=46.2d for Indian product and 27.7 d for Korean product) at the same temperature. The half-lives of the total monoterpenes in biopesticides were ranged from 3.9 d to 27.7 d at $30^{\circ}C$. Among the monoterpenes, the stability ascaridole and p-cymene were decreased in acidic condition. All the bioactive substances showed similar stability on the different thermal conditions. CONCLUSION:The half-lives of most bioactive substance from castor oil and wormseed extracts were less than 100 d. To increase the stability of bioactive substance in biopesticide, stabilizing additives like antioxidant and oxygen remover should be considered to extend of the shelf-life.

• 한국응용과학기술학회지
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• 제7권1호
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• pp.25-30
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• 1990

The seeds of 16 domestic plants were studied for their moisture, total lipids and fatty acid composition. Of the 16 seeds, chestnut, corn, mungbean and ginko nut yielded less than 9% by weight of total lipids compared to others that gave 20-73%. The identified fatty acids from the seed lipids ranged from lauric acid (12:0) to lignoceric acid(24:0). It was intended in this study to classify the seed lipids according to their major fatty acids: Group t-Oleic acid; Group 2-0leic acid and linoleic acid; Group 3-linoleic acid; Group 4-linolenic acid ; Group 5-erucic acid ; Group 6-ricinoleic acid. The saturated fatty acid content of mungbean (33%) was the highest among the seed lipids studied. The highest value for the P/S fatty acid ratio(10) was in perilla.

• KSBB Journal
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• 제14권6호
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• pp.696-701
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• 1999

본 연구에서는 lipase를 이용해서 ricinoleic acid를 대량생산하기 위해 피마자유의 완전가수분해 조건을 찾고자 하였다. 널리 알려진 lipase CR, lipase CC, lipase PP를 대상으로 피마자유의 가수분해의 가능성을 시험하고, 유기용매를 사용함으로써 가수분해도를 향상시킬 수 있음을 확인하였다. 일반적으로 lipase의 활성을 감소시키는 극성용매의 경우 피마자유의 가수분해에 있어서도 효소의 활성을 감소시켰고, 물과 섞이지 않는 hydrophobic solvent가 피마자유의 가수분해도를 크게 증가시키는 것을 확인하였다. 본 연구에서는 isopropyl ether의 효과가 가장 크며, 조건에 따라 가수분해도를 두 배 이상 증가시킨다는 것을 확인하였다. 그리고 유기용매를 사용함으로써 pH의 영향을 바꾸거나 감소시킬 수 있다는 사실도 확인하였다. 용매와 물의 부피비에 의해서 가수분해가 영향을 받는다는 사실과 특히 유기용매보다는 물의 양에 절대적으로 영향받는다는 사실을 발견하였다. 하지만, 물과 유기용매의 부피비와 함께 lipase와 피마자유의 무게비도 매우 중요하다는 것을 확인하였다. 30$^{\circ}C$에서 isopropyl ether를 사용할 경우 무게 비로 2 wt%일 때는 약 82%, 4 wt% 이상의 lipase CC나 lipase CR을 사용하면 피마자유가 완전히 가수분해되는 사실을 발견하였다.