• 환경생물
• /
• 제22권1호
• /
• pp.167-170
• /
• 2004

Phytopathogenic fungi들에 대해 매우 공격적인 Trichoderma hurzianum은 다른 곰팡이들의 세포벽 주성분인 chitin을 분해하는 extracellualr chitinase를 분비하기 때문에 매우 효율적인 biocontrol agent로 사용할 수 있다. Trichoderma harzianum의 생물학적 제어 기능을 이용하기 위하여 extracellualr chitinase의 생산성을 높일 수 있는 batch모드에서의 적정 환경을 찾는 일은 매우 복잡하다. 이러한 문제를 해결할 수 있는 방법이 바로 self-directing optimization이다 이 방법을 이용하여 실험한 결과 6∼7차례의 실험만으로 최적의 온도 혹은 pH 및 폭기율과 교반양을 구할 수 있었다. 그 결과 온도 $32^{\circ}C$, pH4.9, 폭기율 3.22 ι, 교반율 225 rpm에서 chitinase의 최대 생산량 16.825 U를 나타내었다. 이는 온도 $25^{\circ}C$, pH 5, 폭기율 1l, 교반율 150 rpm에서 나타난 chitinase 생산량 4.221 U에 비교하면 3.99배 증가한 효과를 나타내고 있다. 이로서 self-directing optimization방법을 통한 최적화 기술이 효소 생산 기술에 있어 그 최적화 과정을 단축시키고 최종 과정에 도달하는 새로운 기술로 적용할 수 있음을 알 수 있었다.

• Journal of Microbiology and Biotechnology
• /
• 제14권2호
• /
• pp.237-242
• /
• 2004

Cell growth and the production of pullulan by Aureobasidium pullulan HP-2001, the UV-induced mutant of A pullulans ATCC 42023, increased with increased concentration of glucose up to 15.0% (w/v). Maximal production of pullulan in the flask scale was 27.65 g/l, when concentrations of glucose and yeast extract were 15.0 and 0.25% (w/v), respectively. Maximal conversion rate of pullulan from glucose as the carbon source was 0.37, when those of glucose and yeast extract were 5.0 and 0.15% (w/v), respectively. On the basis of total amount of pullulan, the conversion rate of pullulan from glucose, and utilization rate of glucose to cell mass and pullulan by A. pullulans HP-2001, the optimal concentrations of glucose and yeast extract for the mass production of pullulan were determined to be 10.0 and 0.25% (w/v), respectively, at which concentrations the production of pullulan and its conversion rate were 27.14 g/l and 0.27, respectively. Maximal production of pullulan with optimized concentrations of carbon and nitrogen sources by A. pullulans HP-200l in a 7-1 bioreactor was 32.12 g/l for 72 h culture, and that in a 100-1 bioreactor with the inner pressure of $0.4 kgf/cm^2$ was 36.87 g/l. Increased inner pressure of a 100-1 bioreactor resulted in a higher concentration of dissolved oxygen in the medium, which might enhance the production of pullulan by A. pullulans HP-2001.

• Journal of Microbiology and Biotechnology
• /
• 제29권4호
• /
• pp.587-595
• /
• 2019

Pharmacological research on (CHA), a marine-derived quinazolinone alkaloid with significant cytotoxic activity, is restricted by low yields and is a problem that needs to be settled urgently. In this work, the selection of additional nitrogen sources and the optimization of additional concentrations and longer fermentation times using ammonium acetate, were investigated. CHA production was optimized to 62.1 mg/l with the addition of 50 mM ammonium acetate at 120 h of the fermentation in the shaker flask. This feeding strategy significantly increased 3-deoxy-arabino-heptulosonate-7-phosphate synthase activity and transcript levels of critical genes (laeA, dahp, and trpC) in the shikimate pathway compared with the non-treatment group. In addition, the selection of the feeding rate (0.01 and $0.03g/l{\cdot}h$) was investigated in a 5-L bioreactor. As a result, CHA production was increased by 57.9 mg/l with a $0.01g/l{\cdot}h$ ammonium acetate feeding rate. This work shows that the strategy of ammonium acetate supplementation had an effective role in improving CHA production by Aspergillus fumigatus CY018. It also shows that this strategy could serve as an important example of large-scale fermentation of a marine fungus in submerged culture.

• 한국생명과학회:학술대회논문집
• /
• 한국생명과학회 2006년도 제47회 학술심포지움 및 추계국제학술대회
• /
• pp.111.2-111.2
• /
• 2006

• Journal of Microbiology and Biotechnology
• /
• 제15권1호
• /
• pp.7-13
• /
• 2005

Chitosan-alginate capsules were formed by electrostatic interactions and exhibited an appropriate mechanical strength, permeability, and stability for the culture of hepatocytes. Pig hepatocytes were isolated and hepatocyte spheroids formed and immobilized in chitosan-alginate capsules. An encapsulation procedure of 3 min and spheroid formation period of 24 h were the optimum conditions for the best liver functions. Pig hepatocytes with a cell density of $6.0{\tomes}10^6$ cells/ml in the capsules were found to be most suitable for application in a bioartificial liver support system. The encapsulated pig hepatocyte spheroids exhibited stable ammonia removal and urea secretion rates in a bioreactor for 2 weeks. Accordingly, chitosan-alginate encapsulated hepatocyte spheroids in a packed-bed bioreactor would appear to have potential as a bioartificial liver.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제7권3호
• /
• pp.163-170
• /
• 2002

This report describes the use of a transtubular bioreactor to study the relative effects of diffusion versus perfusion of medium on antibody production by a hybridoma cell line. The study was performed with a high-density cell culture maintained in a serum-free, low-protein medium for 77 days. It was determined that the reactor possessed a macro-mixing pattern residence time distribution similar to a continuous stirred tank reactor (CSTR), However, due to the arrangement of the medium lines in the reactor, the flow patterns for nutrient distribution consist of largely independent medium path lengths ranging from short to long. When operated with cyclic, reversing, transtubular medium flow, some regions of the reactor (with short residence times) are more accessible to medium than others (with long residence times). From this standpoint, the reactor can be divided into three regions: a captive volume, which consists of medium primarily delivered via diffusion; a lapped volume, which provides nutrients through unilateral convection; and a swept volume, which operates through bilateral convection. The relative sizes of these three volumes were modified experimentally by changing the period over which the direction of medium flow was reversed from 15 min (larger captive volume) to 9 h (larger swept volume). The results suggest that antibody concentration increases as the size of the diffusion-limited (captive) volume is increased to a maximum at around 30 min with a sharp decrease thereafter. As reflected by changes in measured consumption of glucose and production of lactate, no significant difference in cellular metabolism occurred as the reactor was moved between these different states. These results indicate that the mode of operation of the transtubular bioreactor may influence antibody productivity under serum-free, low-protein conditions with minimal effects on cellular metabolism.

• Journal of Microbiology and Biotechnology
• /
• 제9권4호
• /
• pp.443-449
• /
• 1999

Bacillus subtilis BK-17 which produces a novel protease with fibrinolytic activity was isolated from soybean paste. Bioreactor production of the enzyme was studied in order to optimize fermentation conditions such as medium concentration, pH, agitation speed, and temperature. Under most cultural conditions, enzyme production initially began when the cell growth stopped. The onset of the enzyme production was indicated by rapid increase in both dissolved oxygen (DO) and pH. Two- to three-times more concentrated medium than the flask optimum medium yielded higher enzyme production in the bioreactor fermentation. When the medium pH was controlled constant, pH 6.5 exhibited the highest activity in the range of 6.0 to 7.5, but the activity was similar to the case when the pH was initially adjusted to 7.5 and subsequently maintained within a relatively wide range of 6.4 to 7.8. Agitation speed did not affect the enzyme production with an exception of DO reaching zero. Fermentation time was reduced when temperature increased within the range of $25^{\circ}C$ to$37^{\circ}C$. However, the highest activity, along with the slow decrease of the enzymatic activity after reaching the maximum value, was observed at $25^{\circ}C$. By shifting the temperature from $37^{\circ}C$ to $25^{\circ}C$immediately after DO reached the minimum level, the high enzyme production of 1,100 U/ml along with the short fermentation period of 13 h could be obtained.

• Environmental Engineering Research
• /
• 제21권2호
• /
• pp.196-202
• /
• 2016

Membrane bioreactor (MBR) technology has previously been used by water industry to treat high salinity wastewater. In this study, an anoxic-oxic biofilm-membrane bioreactor (AOB-MBR) system has been developed to treat mustard tuber wastewater of 10% salinity (calculated as NaCl). To figure out the effects of operating conditions of the AOB-MBR on membrane fouling rate ($K_V$), response surface methodology was used to evaluate the interaction effect of the three key operational parameters, namely time interval for pump (t), aeration intensity ($U_{Gr}$) and transmembrane pressure (TMP). The optimal condition for lowest membrane fouling rate ($K_V$) was obtained: time interval was 4.0 min, aeration intensity was $14.6 m^3/(m^2{\cdot}h)$ and transmembrane pressure was 19.0 kPa. And under this condition, the treatment efficiency with different influent loads, i.e. 1.0, 1.9 and $3.3kgCODm^{-3}d^{-1}$ was researched. When the reactor influent load was less than $1.9kgCODm^{-3}d^{-1}$, the effluent could meet the third discharge standard of "Integrated Wastewater Discharge Standard". This study suggests that the model fitted by response surface methodology can predict accurately membrane fouling rate within the specified design space. And it is feasible to apply the AOB-MBR in the pickled mustard tuber factory, achieving satisfying effluent quality.

• 미생물과산업
• /
• 제19권4호
• /
• pp.14-26
• /
• 1993

Optimizing protein production in recombinant E. coli strains involves manipulation of genetic and environmental factors. In designing a production system, attention must be paid to gene expression efficiency, culture conditions and bioreactor configuration. Although not much emphasis was given to the physiology of host strains in this review, an understanding of the relationship between the physiology of host cell growth and the overproduction of a cloned gene protein is of primary importance to the improvement of the recombinant fermentation processes. Sometimes it is desirable to make use of gene fusion systems, e.g. protein A, polypeptide, gutathione-S-transferase, or pneumococcal murein hydrolase fusion, to facilitate protein purification.

• 한국미생물·생명공학회지
• /
• 제48권3호
• /
• pp.344-357
• /
• 2020

Streptococcus zooepidemicus 유래의 세포외 고분자물질인 히알루론산(hyaluronic acid) (HA)을 대량 생산하기 위해, 균주 개량, 생산배지 및 배양공정 개발에 관한 연구를 수행하였다. HA 고생산성 변이주를 선별하기 위해 약 99%의 사멸률을 보이는 ethylmethane sulfonate (EMS) 처리조건을 적용해서, 지속적인 random screening 방법으로 고생산성, 고안정성의 변이주들을 선별할 수 있었다. HA를 고농도로 생산하기 위해서는, 이 균주의 생화학 및 배양생리적 특성에 기반한 최적 배지개발이 필수적이라고 판단하여, one-factor-at-a-time (OFAT), full factorial design (FFD), steepest ascent method (SAM) 및 response surface method (RSM) (반응표면분석법)을 순차적으로 적용하여 통계적 배지 최적화 실험을 수행하였다. 최적 배지조성에서 플라스크 배양에 의한 HA 생산성은 5.38 g/l로서, 이전 배지(3.54 g/l)에 비해 약 52% 향상된 생산량을 얻을 수 있었다. 또한 선별된 우량균주와 최적화된 생산배지를 이용하여 5 L 발효조에서 배양공정 최적화 연구를 수행하였다. 이 균주의 생리학적 특성을 고려할 때, HA 생산성을 높이기 위해서는 (배양 중 HA 축적으로 인해 고점도를 띠는) 배양액으로의 충분한 용존산소 공급이 매우 중요한 요인인 것으로 판단되었다. 따라서 용존산소 공급과 밀접하게 관련있는 발효조의 교반시스템(교반 날개 종류, 크기 및 배치 등) 및 교반속도에 대한 최적화 연구를 수행하였다. 그 결과, 교반축 하부에는 Rushton turbine-type, 상부에는 marine-type의 확장된 교반날개(기존 대비 직경 1.3배 확장)가 설치된 경우, 450 rpm에서 강화된 혼합력과 충분한 용존산소 공급으로 인해 HA 생산성이 기존 플라스크 배양 대비 약 1.8배(9.79 vs. 5.38 g/l) 더 높은 것으로 확인되었다. 최종적으로 HA 배양공정의 scale-up 가능성을 확인하기 위해, pilot 규모의 50 L 발효조 배양을 최대 300 rpm의 교반속도에서 수행하였다. 처음으로 시도한 50 L 배양임에도 불구하고, HA 최대 생산성 면에서 볼 때, 5 L 발효조 결과와 거의 동일한 수준(98.5%) (9.11 vs 9.25 g/l)의 생산량을 얻을 수 있었다. 반면 지수기 성장단계인 배양 15시간까지의 50 L 배양의 HA 평균생산속도(r_p)는 0.46 g/l/hr로서 0.62 g/l/hr인 5 L 배양 대비 약 74% 정도에 머무는 것으로 나타났다. 따라서 생산 발효조의 scale-up 시, 생산균주의 전단응력 민감성(shear damage)을 함께 고려하면서, 산소전달계수(k_La)를 기반으로 하는 교반시스템에 대한 체계적인 연구가 진행된다면, HA 생산속도도 증가될 수 있는 긍정적인 결과를 얻을 수 있을 것으로 기대된다.