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한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.315-321
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Rabbit anti-bovine heart PDH antiserum was raised against El(a, b) isolated from PDC, and then applied to detect Ela and Elb. Appropriate amounis of El were fractionated by SDS-PAGE and electrophoretically transferred to nitrocellulose membrane. The Ela and Elb on the membrane were incubated with anti-El antiserum and identified by GAR-HRP system. It has been found that the immunodetection sensitivity of Ela and Elb were directly proportional to the amount of antigen and transfer time. The lengthy transfer times increased the immunodetection sensitivity of Ela and Elb. The maximal detection sensitivity of Western blotting of Ela and Elb was achieved at 3.5 V/cm for 16-hour transfer under these experimental conditions.
곡물류의 형질전환에 관한 연구.II. Electroporation에 의해 벼 원형질체로 도입된 유전자의 발현
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.323-327
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Protoplasts isolated from embryogenic cell suspensions were electroporated in buffered solutions containing plasmid DNA of pBI121. Transient GUS (beta-glucuronidase) activity measurement and selection for kanamycin resistent showed that expression of foreign genes and stable loransformation were achieved. GUS transient gene expression was increased by increasing DNA concentration of pBI121 plasmid and affected by the level of the applied voltage. An optimal level of GUS activity was obtained after electroporation with a pulse of 200 voltage/1180 uF. Protoplast viability was up to the 60% at the optimal voltage. Cell colonies resistent to 200ug/ml kanamycin were selected in agar medium and identified by histochemical GUS assay.
역원추형 발효조에서의 동시당화발효에 의한 전분으로 부터의 연속 에탄올 발효
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.329-334
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In an attempt to develop a novel process for ethanol production from starch, a simultaneous saccharification and fermentation (SSF) process using Zymomonas mobilis and amyloglucosidase (AMG) was studied in continuous modes. Compared with a conventional cylindrical column type of fermentor, the tapered column type of fermentor was found to be superior in terms of reactor performance for ethanol fermentation. The tapered columm fermentor packed with coimmobilized Z. mobilis and AMG alleviated the problems which were associated with CO2 evolution and provided a significantly better flow pattern for both liquid and gas phases in the fermentor without channelling. However, the fluidized bed type of tapered column fermentor using flocculent strain of Z. mobiles and immobilized AMG showed lower productivity (5.2g/1/h) than that of packed bed type of tapered column fermentor(9.2g/l/h).
균체재순환 및 동시당화발효에 의한 전분으로 부터의 반회분식 에탄올 발효
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.335-339
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전분으로 부터 에탄올을 생산하기 위한 경제적인 공정을 개발하기 위하여 Zymomonas mobilis와 당화효소(AMG)를 사용한 반 회분식 동시 당화 발효공정올 연구 하였다. 응집성 에탄올 균주인 Z. mobilis ZM40l과 침전조를 사용한 균체 재순환 방식에 의한 반회분식 동시 당화발효 공정에서는 에탄올 생산성이 제2차 및 제3차 발효에서 각각 4.1g / I / h 및 4.3 g / I / h이었다. 이에 비해 미세여과막(microfiltration) 장치에 의한 Z. mobilis ZM4의 재순환 방식을 사용하는 공정에서는 에탄올 생산성이 제2차 및 제3차 발효에서 모두 5.4 g / l / h로 더 높았다. 에탄올 생산 시설이 large-seale임을 고려할 때 미세여과막을 사용하는 반회분식 공정이 에탄올 생산성과 seale-up의 용이성 및 운전의 간편성등의 관점에서 가장 개발 가능성이 높은 공정인 것으로 판단되었다
In order to develop economic processes for ethanol production from starch, a simultaneous saccharification and fermentation(SSF) process using Zymomonas mobilis and amyloglucosidase (AMG) was studied in semibatch modes using cell recycle. The cell recycle was carried out by adopting two different methods; microfiltration and settling. The cell recycle using microfiltration revealed higher productivity(5.4 g/l/h) than that using a settler(4.3 g/l/h). Taking the large-scale ethanol fermentation into account, the semibatch process using microfiltration system appeared most promising among others with respect to ethanol productivity, feasibility of scale-up and simplification of operation.
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.341-345
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잣기름과 호도기름의 트리글리세리드 조성을 규명하기 위하여 HPLC의 Tri-analysis column을 사용 분석한 결과는 다음과 같다. 1. 잣기름중 트리글리세리드 조성은 총 14종이 함유되었으며 탄소수별로 확인된 종류가 10종이고, 미확인수는 4종이고, 그 포함 순서는 C52=31.55%, C44=20.72%, C54=17.52%, C34=15.24%, C56=12.50%의 순으로 조성되어 있다. 2. 호도기름중 트리글리세리드 조성은 총 12종이 함유되어 있으며 탄소수별로 확인된 종류가 10종이고, 미확인된 종류는 2종이고, 그 포함 순서는 C44=38.66%, C52=29.47%, C54=16.53%, C34=11.12%, C50=1.51% 순으로 조성되어 있다.
The composition of triglyceride in the oil of walnut and pine-nut were analysed quantitatively by Tri-analysis column of HPLC. The results were as follows: 1. The compositions of triglyceride of pine-nut oil were C52=31.55%, C44=20.72%, C54=17.52%, C34=15.24%, C56=12.50% in order, respectively. 2. The compositions of triglyceride of walnut oil were C44=38.66%, C52=29.47%, C54=16.53%, C34=11.12%, C50=1.51% in order, respectively
Daucus carota 현탁 배양에서 배지 조성 및 세포 응집 크기가 Carotenoid 생합성에 미치는 영향
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.347-353
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The effects of nutrients and cell aggregate size on the cell growth of D. carota and the biosynthesi of carotenoid were investigated. Highest carotenoid content was obtained with sucrose as a carbon source and the equal ratio of ammonium to nitrate. High phosphate concentration stimulated the carotenoid biosynthesis in D. corota. 2,4-D inhibited the cell growth but stimulated the specific carotenoid content at high concentration. By modifying the medium composition based on these findings, three times higher specific carotenoid content and 2.5 times higher total carotenoid content were obtained as compared with the results obtained with basic MS media. Biosynthesis of carotenoid was found to be affected by cell aggregate size; high carotenoid production was obtained from the large aggregated cells resulted from high sucrose concentration.
SCP 생산용 메탄올 자화균주의 연속배양에 의한 배지 최적화
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.355-363
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서울 인근의 생활하수에서 메탄올을 이용하는 박테리아를 분리하였다. 이의 최적 성장을 위한 온도 및 pH는 각각 33℃ 및 7.1이었다. 최대비성장율은 0.42hr-1을 나타내었다. 최소 배지조성은 기본배지의 양을 안정상테에서 연속발효법으로 구하였으며, 그 조성은 다음과 같다(g / l); Methanol 40, 최적 배지 조건에서 구한 최대 세포생산성은 3.8g / l / hr였으며, 이때의 회석율은0.23hr-1 이었다. 최대 세포농도와 그대의 단백질의 함량은 각각 19.5g / l 및 70%였으며, 이때의 회석율은 0.1hr-1이었다
Methanol-utilizing bacterium isolated from sewage samples in Seoul showed optimal temperature and pH of 33℃ and 7.1 for growth, respectively. The maximum specific growth rate was 0.42hr-1. The minimum medium composition was reconstituted depending on the surplus and the deficit of each component in the basal medium at steady state. The optimal composition was given as(g/l); Methanol 40, Under the continuous culture with optimum medium the maximum cell productivity was 3.8g/1/hr at dilution rate . Maximum cell concentration and its protein content were 19.5g/l and 70% at dilution rate of 0.1hr-1, respectively.
농축배지에서 Glucose와 Glutamine 농도가 하이브리도마 증식과 간염표면항원에 대한 단일클론항체 생산에 미치는 영향
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.365-371
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To improve the growth of mouse hybridoma 2c3.1 secreting anti-Hepatitis B surface antigen monoclonal antibody (anti-HBsAg MAb), we had constructed an enriched medium and observed the effects of fetal bovine serum and serum-free supplements including human serum albumin, 'insulin and transferrin', and monoethanolamine. For further enhancement of growth, the concentrations of two major energy sources, glucose and glutamine, were strengthened with various ratios in the enriched medium. Maximum cell growth and monoclonal antibody production obtained in various ratios of glucose/glutamine with an inoculation concentration of 2x105 cells/ml were 0.73x106-4.62x106 cells/ml and 65.1-422.6 ug/ml, respectively. Glutamine was round to be a major energy source and a limiting nutrient in comparison to glucose for 2c3.1 cell cultivation in enriched media with low serum.
Enhancing Effect of Egg Albumin on Ethanol Production and Its Function
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.373-376
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Saccharomyces sake를 이용한 알콜발효에서 배지의 첨가물로 egg albumin 및 phoshatidylcholine이 최종 알콜 농도에 미치는 영향과 그의 기작에 대햐여 분석하였다. Phosphatidycholine-egg albumin이 첨가될 때 최종 알콜 농도는 120g / l 정도 얻어졌으며, 첨가물이없는 경우에 비해 대략 30% 정도, 첨가물 linoleic acid-ergosterol에 비하여 20% 정도 증가되었다. 최종 알콜농도가 높을수록 균체의 알콜내성도 비례적으로 증가하였으며, 두 첨가물 사이의 차이는 지방산보다는 albumin과 ergosterol의 영향으로부터 주로 비롯되었다. 열반성 혹은 가수분해된 albumin을 첨가한 결과로 부터 albumin은 균체외부에 보호막을 형성하기보다는 영양분으로 균체내에 흡수되어 균체성질을 변화시켜 알콜내성을 증진시키며, 이로부터 궁극적으로 최종 알콜 농도를 증가시키는 것으로 판단되었다
In ethanol fermentations with Saccharomyces sake, phosphatidylcholine-egg albumin as a supplement in fermentation media was much more effective in enhancing ethanol production than linoleic acid-ergosterol. It came from the differences in alcohol-tolerance between egg albumin and ergosterol. The egg albumin was supposed to function as a nutrient rather than to form protective layers around the cells against ethanol.
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.377-382
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The characteristics controlled drug release have been studied for biodegradable polymer matrix. Polymer matrix was prepared from glycerine, prednisolone and dextran. Mathematical analysis of the data showed that the release behavior actually conformed to the Higuchi's diffusion controlled model. The release time was increased as drug loading doses increased, whereas decreased as glycerine concentration increased. The release rate did not change by varying molecular weight of dextran.
주정증류 폐기물의 당화 및 구연산 발효에 관한 연구(I)
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.383-390
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주정공장에서 배출되는 쌀보리 주정증류 폐기물을 효소와 산으로 가수분해하여 구연산 발효를 행한 결과를 요약하면 다음과 같다. 폐기물을 농축하여 당의 농도를 50g / l로 하였을때 글루코오즈, 구연산 및 균체농도는 각각 5.7g / l, 0.8g / l 및 3.86g / l가 얻어졌고 a- 및 B-amylase에 의한 폐기물의 효소 가수분해에 의해서 글루코오즈, 구연산 및 균체농도가 각각 81g / l, 1g / l 및 5.33g / l가 얻어졌으며 25% 염산에 의한 산가수분해에 의해서는 각각 21.5g / l, 1.75g / l 및 4.9g / l가 얻어졌다. 10시간동안 계속된 2차 산 가수분해를 행한 결과 글루코오즈의 농도가 3.44g / l로 장기간의 산 가수분해는 생성된 글루코오즈를 분해시키는 속도가 3당류 이상의 올리고 환원당의 분해속도보다 빨랐다. 폐기물내 균체 접종량을 0.5%에서 2%로 증가시킴에 따라 균체농도는 1.35g / l에서 3.71g / l 로 증가 하였으나 구연산 농도는 0.4g / l에서 0.12g / l로 감소하였으며 또한 질산암모늄을 3g / l를 첨가할 경우 균체 농도는 3.7g / l에서 7.3g / l로 증가하였으나 구연산 농도는 0.12g / l에서 0.08g / l로 감소되었다. MnSO4ㆍ45H2O의 농도가 0.0125g / l 일때 구연산 생산량이 가장 많았으나 농도가 증가함에 따라 구연산 생산이 감소하였으므로 금속이온이 과다하게 존재할때는 구연산 생산이 저해됨을 알았다
Alcoholic distillery waste was utilized as dual purposes to produce citric acid and to reduce the amount of waste to be treated. Enzyme and acid hydrolysis of this waste were studied to suggest effective way of present purpose. Enzymatic hydrolysis of this naked barley alcoholic distillery waste by a-and B-amylase gave glucose as 8g/l concentration at for 6 hours, which produced 1g/l citric acid and 5.33g/l mycelial. This waste material hydrolyzed with 25% HCl at showed 21.5g/l glucose and produced 1.75g/l citric acid with 4.9g/1 mycelial. The glucose concentration was decreased to 3.44g/l by further 2nd acid hydrolysis because the monosugars were decomposed at prolonged hydrolysis conditions. The addition of 3g/l NH4NO3 increased the mycelial growth but reduced the amount of citric acid formed. The formation of citric acid was increased at low concentration of manganese ion.
인간 신장세포로부터 새로운 배양공법에 의한 scu-PA의 생산성 향상
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.391-396
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4.010-3 (IU/cells/day) of maximum specific scu-PA production, which was higher than those by a 75 ㎠T-flask and conventional perfusion systems, was maintained by tube-type bioreactor at 0.35 (1/h) of perfusion rate corresponding to 0.15 (dynes/) of shear stress. The production of scu-PA is also increased as shear stress was slowly increased, which is similar to natural human blood circulation. The tube reactor proves that there may be no limitation of oxygen supply by showing 1.0 (1/h) of oxygen transfer coefficient at steady state and this system yields much lower shear stress of 0.3(dynes/ ㎠) than that of 5-8(dynes/ ㎠) by conventional agitation systems.
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.397-402
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Hydrolysis of inulin was investigated employing various commercially available resin catalysts for the production of high grade fructose syrup. The particle size and porosity of the resin significantly affected the distribution of the products, indicating that the intraparticle diffusion of reactants controls thc selectivity as well as the reaction rate. To confirm the effect of the intraparticle diffusion, two different types of resin catalysts were prepared: the one having sulfonic acid group distributed uniformly throughout genular microparticles (A-type) and the other having sulfonic acid group located mainly at the exterior surface of genular microparticles (E-type). The results were found that the reaction rate and the selectivity of the E-type catalyst were higher than those of the A-type catalyst.
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.403-410
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Application of the Rheocatch Hotwire Monitoring System for food biotechnology process was evaluated. The growth of microogranism, E coli (JM 83 and Sigma) and Corynesccfertun glutamicum, were monitored. in the fermentor. The cell growth could not be detected the temperature differences between the hotwire and samples(ΔT) as indicated by the monitoring system during the fermentation processes. The cell concentration of less than 2g/dl was not sufficient to generate the measurable temperature difference in the fermentor. In order to calibrate the Rheocatch Monitoring System, the temperature difference as a function of solute concentration (microbial cells, sodium cholide, sucrose and dextran) was studied. The relationship between ΔT and the concentration of microbial cells, sucrose and dextran can be expressed in a power series. Further studied with dextran indicated that viscosity and/or kinematic viscosity increase exponentially with an increase in ΔT This is regardless of the concentration and molecular weight of dextran. ΔT linearly increases with the logarithm of molecular weight, while the logarithm of viscosity and the logarithm of kinematic viscosity increase with the logarithm of molecular weight.
역미셀을 이용한 Alkaline Protease의 추출분리
한국생물공학회 KSBB Journal 제5권 제4호 1990.12 pp.411-414
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The extraction behavior of an alkaline protease using reversed micelles was investgated. The reversed micellar solution consisted of AOT in isooctane. It was found that distribution of arkaline protease into the organic phase increased at lower pH, lower ionic strength, and higher AOT concentration. When the real fermentation broth was extracted of alkaline protease, an activity yield of 20% and a purification factor of 2.0 were obtained.
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