• Biotechnology and Bioprocess Engineering:BBE
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• 제10권2호
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• pp.142-148
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• 2005

This paper proposes a cell age model for Penicillium chrysogenum fed-batch cultivation to supply a qualitative insight into morphology-associated dynamics. The average ages of the segregated cell populations, such as growing cells, non-growing cells and intact productive cells, were estimated by this model. A combined model was obtained by incorporating the aver-age ages of the cell sub-populations into a known but modified segregated kinetic model from literature. For simulations, no additional effort was needed for parameter identification since the cell age model has no internal parameters. Validation of the combined model was per-formed by 20 charges of industrial-scale penicillin cultivation. Meanwhile, only two charge-dependent parameters were required in the combined model among approximately 20 parameters in total. The model is thus easily transformed into an adaptive model for a further application in on-line state variables prediction and optimal scheduling.

• Journal of Microbiology and Biotechnology
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• 제16권4호
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• pp.590-596
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• 2006

Clavulanic acid (CA) produced by Streptomyces clavuligerus is degraded during the bacterial cultivation. The degradation was examined in three different aspects, including physical, chemical, and enzymatic effects, in order to understand the degradation during the cultivation. The result showed that CA was unstable in the production medium containing ammonium salts and amino acids, owing to ammonium ions and amine groups. In addition, the degradation was not only due to instability of CA by metabolites and proteins, but also enzymes from S. clavuligerus such as $\beta-lactamase$ and penicillin-binding proteins. However, the degradation caused by these enzymes was not highly significant compared with the degradation during the cultivation, owing to irreversible reactions between CA and enzymes.

• Journal of Plant Biotechnology
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• 제42권2호
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• pp.111-116
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• 2015

Interest and great demand for blueberry (Vaccinium corymbosum) have increased, as V. corymbosum is now one of the most economically important crops in Korea. It is expected that blueberry production and the area planted for cultivation will increase consistently in the years ahead because of high profitability and the consumer's demand for healthy ingredients. Effective mass production of blueberry is urgently needed for commercial cultivation establishment, but a main limitation is lack of a propagation system that produces a disease-free plant material for commercial plantation. A large amount of research has focused entirely on developing tissue culture techniques for blueberry propagation. However, controlling fungal and bacterial contamination of woody plant material is extremely difficult. Our study was conducted to investigate the effect of biocide addition during the in vitro culture of blueberry on plantlet growth and contamination occurrence. Four biocides, including Plant Preservative Mixture ($PPM^{TM}$), vancomycin, nystatin and penicillin G, were used in varying concentrations during the in vitro propagation of blueberry. When nystatin was added into the medium at low concentrations, the overall growth of blueberry plantlets was retarded. Addition of vancomycin and penicillin G in high concentrations decreased contamination but induced plantlet mortality. On the other hand, when 1ml/L $PPM^{TM}$ was added, the growth characteristics of blueberry plantlets did not significantly differ from non-treatment (control), and the contamination occurrence rate was very low. From these results, we found that the addition of the appropriate biocide could provide an effective method to reduce contamination in the culture process, thereby raising in vitro production efficiency.

• 한국식품위생안전성학회지
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• 제30권1호
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• pp.51-58
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• 2015

본 연구는 들깻잎과 들깻잎 생산환경을 대상으로 31개의 시료를 채취하여 S. aureus 를 분리 하였다. 분리된 S. aureus 31주의 toxicity를 평가하고자 독소유전자와 항생제 내성을 검색하였다. 그 결과 분리된 균주에서 4개의 서로 다른 독소유전자 패턴은 확인하였으며 sea와 sed유전자를 동시에 보유하는 균주는 4균주 (12.9%)였고, sed 유전자를 보유하는 균주는 9균주 (29.0%)였으며 see 유전자를 보유하는 균주는 1균주 (3.2%)였다. 한편 seb와 sec 유전자를 보유하는 균주는 없었다. 항생제 내성평가결과, 12제의 항생제(penicillin, ampicillin, oxacillin, amoxicillin-clavulanic acid, cefazolin, cephalothin, imipenem, gentamicin, tetracycline, ofloxacin, norfloxacin, and erythromycin)에 내성을 보인 균주는 7균주 (22.6%)였다. 또한 분리된 균주의 2균주 (2.6%)는 5제의 항생제(penicillin, ampicillin, amoxicillin-clavulanic acid, gentamycin, and telithromycin)에 내성을 보였고 MRSA (Methicilline Resistant Staphylococcus aureus)는 포장지, 장갑, 들깻잎에서 발견되었다. 따라서 본 연구결과는 들깻잎에 오염된 S. aureus에 의하여 독소형 식중독이 발생할 가능성을 시사하며 들깻잎과 생산 환경에서 항생제 저항성 S. aureus가 검출되어 의약계뿐만 아니라 농업현장에서도 항생제내성균주 출현을 예방하는 대책이 요구된다.

• 한국미생물·생명공학회지
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• 제9권1호
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• pp.29-34
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• 1981

대장균으로부터 페니실린 아미다제의 생산을 최적화하기 위하여 배지조성 및 발효 중 효소생산에 미치는 제반영향들에 대하여 연구하였다. 최적배지조성은 3.5% 트립톤, 1.5% 글루타민산 소듐, 0.5% 이스트추출물 그리고 0.15%의 페닐초산이었다. 효소생산 유도물질로서 0.15%의 페닐초산을 발효초기에 가해줄 때 효소생산성이 약5배 증가하였다. 최적화된 배지 및 발효조건에서 발효 16시간만에 최대의 효소생산을 나타내었고 이때의 생산성은 약 102.5 units/l 발효액이었다. 이러한 최적조건에서 발효할 때 현재까지 보고된 효소생산성과 비교하여 상당한 생산능력의 향상을 보였다.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.231-236
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• 2004

The inexpensive large-scale production of pure PGA (Penicillin G Acylase) has been a commercial goal. PGA has been used as a model enzyme in the development of simple one-step purification methods. In this study, the purification of poly-His tagged PGA protein secreted into the periplasmic space was carried out by using immobilized metal-ion affinity chromatography (IMAC). The PGA gene was obtained from E. coli ATCC 11105. Codons encoding histidines were fused at the C-terminus of the PGA gene by PCR. E. coli JM109 harboring pPGA-HIS6 vector produced active his-tagged acylases in the presence of lac promoter during cultivation at $26^{\circ}C$. The maximum specific activity of the acylase purified by using one-step chromatography after osmotic shock was 38.5 U/mg and was recovered with the yield of 70％. Both 23 kDa ($\alpha$) and 62 kDa ($\beta$) subunits were recovered by using IMAC with just C-terminus tagging of the $\beta$ subunit. The purification of the periplasmic fraction by osmotic shock and that of purified acylase was increased by 2.6-fold and 19-fold, respectively, compared to the crude extract.

• 한국미생물·생명공학회지
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• 제27권5호
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• pp.399-403
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• 1999

For the production of trehalose, microorganisms capable of producing trehalose extracellularly were screened from the stock cultures in our laboratory. among them, Micrococcus luteus IFO 12708 showed the highest productivity of trehalose. For the increase of productivity, the mutant strai Hs-208 having higher trehalose production was selected with NTG(N-methyl-N'-nitrosoguanidine) mutagenesis, which led to the decrease of the specific activity of trehalose phosphorylase(3.2-fold) as compared to the wild strain. The optimum condition for the trehalose production was established as follows: 20g/l of glucose and 6g/l of tryptone were used as a sole carbon source and nitrogen source, respectively, and cultivations were carried out at 3$0^{\circ}C$ and pH 6.0. After 20hrs cultivation, addition of 20unit/ml penicillin G led to the higher conversion yield of trehalose. Under the optimum condition, 6.547g/l trehalose was produced with conversion yield of 32.7%.

• 한국미생물·생명공학회지
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• 제3권1호
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• pp.7-15
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• 1975

The possibilities of utilizing acid-hydrolyzate of "Sliced and dried sweet potatoes" as a carbon source for the microbial production of L-Glutamic Acid(L-GA) with Micrococcus glutamicus were investigated and the results showed as follows： 1) The highest hydrolysis rate, 74.6% of the reducing sugar based on the weight of dry matter, was obtained when the sweet potatoes were hydrolyzed with 0.8% of HCI at 2.0kg/$cm^2$ for 30 minutes. The most favorable hydrolyzate for the growth of the cells, however, was found to be the one obtained by treating the sweet potatoes with 0.5% HCI at 2. 0kg/$cm^2$ for 10 minutes. Reducing sugar content of the hydrolyzate was 10% as glucose. 2) Biotin content of the hydrolyzate was 25$\mug$/1 and it was proved to be excess in amount for the L-GA production. 3) The effects of addition of antibiotics, alcohols and fatty acid esters on the L-GA production were tested in the biotin excess medium. The production of L-GA was most increased to 32.5g/l with the addition of 10 I. U. of penicillin per ml. to the culture medium at 4 hours after inoculation. But the addition of alcohols, especially fatty acid esters, showed no significant effects. 4) Among the organic nutrients tested. " Gluten acid hydrolyzate" greatly enhanced the production of L-GA adding it＇s concentration of 1.0% to the medium. 5) The maximum production of L-GA resulted in 35g/1 when the cells were grown for 48 hours in the hydrolyzate medium supplemented with 1.0% of "Gluten acid hydrolyfate" and with 10 I. U. of penicillin per ml added at 4 hours after cultivation.

• 한국식품과학회지
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• 제43권2호
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• pp.134-141
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• 2011

본 연구는 들깻잎과 들깻잎 생산환경을 대상으로 276개의 시료를 채취하여 B. cereus를 분리 하였다. 분리된 B. cereus 200주의 toxicity를 평가하고자 독소유전자와 항생제내성을 검색하였다. 그 결과 분리된 균주에서 11개의 서로 다른 독소유전자 패턴은 확인하였으며 5개의 설사형독소와 구토형 독소를 모두 생성할 수 있는 균주는 21%였다. 가장 빈번하게 검출되는 독소유전자는 nheA(100%), enFM(100%), hblA, C, D(66.5%)였으며 EM은 가장 낮은 빈도(21.0%)로 검출되었다. 항생제 내성평가결과 분리된 대부분의 B. cereus는 18종의 항생제 중 10개의 항생제에 대해서는 감수성이었으나 ${\beta}$-lactam계 항생제인 penicillin(100%), ampicillin(100%), oxacillin(94.9%), amoxicillin-clavulanic acid(95.6%), cefazolin(78.2%)과 비${\beta}$-lactam계 항생제 rifampicin(58.0%)에 대해서 저항성을 보이는 것으로 나타났다. 들깻잎과 들깻잎 생산환경에서 분리된 B. cereus의 독소유전자와 항생제내성 패턴은 서로 유사하였다. 따라서 본 연구결과는 들깻잎에 오염된 B. cereus에 의하여 설사형 뿐만 아니라 구토형 식중독이 발생할 가능성을 시사하며 들깻잎과 생산환경에서 항생제 저항성 B. cereus가 검출되어 의약계뿐만 아니라 농업현장에서도 항생제내성균주 출현을 예방하는 대책이 요구된다.

• 한국미생물·생명공학회지
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• 제25권4호
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• pp.339-345
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• 1997

Under the light intensity of 25-72$\mu$E/ m$^{2}$/s Botryococcus braunii UTEX 572 grew faster than Botryococcus sp. GE 24 isolated from a freshwater lake. The specific growth rate ($\mu$) of B. braunii UTEX 572 was highest at 0.260 (1/day) on a dry weight basis in Chu 13 medium from 1 to 9 days of incubation and then continuously decreased. Carbohydrate concentration and cellular nitrogen and phosphorus contents of B. braunii UTEX 572 gradually decreased with light intensity over a range of 25-72$\mu$E/m$^{2}$2/s, whereas the concentrations of protein and cellular N:P ratio increased with light intensity. Chlorophyll-a concentration showed a decreasing tendency with light intensity. The dry weight of B. braunii UTEX 572 increased in the highest rate of 83 mg/l/day at pH 8.0. When the N: P ratio of Chu 13 medium was adjusted to 50: 1 by addition of nitrogen source, dry weight increasing rate was 115 mg/l/day between 20 and 28 days of incubation which was the highest value during the cultivation. Cell growth in an open culture of B. braunii UTEX 572 was highest with Chu 13 medium, whereas that with Chu 13 medium adjusted to pH 7.0, containing 250 mg/l penicillin, or containing 1% glucose was reduced on a large scale. However, this result shows the possibility of the mass cultivation of B. braunii UTEX 572 in an open system competing with other microorganisms.