• KSBB Journal
• /
• 제16권6호
• /
• pp.527-532
• /
• 2001

공업용 유기용매나 그리스 제거제, 전자기판 세척제로 널리 사용되어 온 TCE는 적절한 처리 방법이 개발되지 못하여 환경오염을 일으키고 있으며, 유전자변이 및 암을 유발하는 것으로도 알려져 효율적인 처리기술 개발이 요구된다. 미생물막을 이용한 처리 방법은 TCE를 무해한 최종산물로 분해시킬 수 있으며, 운전비 및 설비비도 낮아 경쟁력 있는 처리 기술로 평가되고 있다. TCE 자체는 미생물의 성장기질로 사용되지 못하므로 공동대사 과정을 거쳐 분해된다. TCE 및 TCE 분해산물의 높은 독성으로 인하여 TCE 분해효소 및 세포 분해산물의 높은 독성으로 인하여 TCE 분해효소 및 세포 불활성화가 일어나서 장기간 안정된 반응기 운전이 어렵다는 단점과 성장기질을 공급하는 경우 유발되는 경쟁저해로 인한 분해효율 감소를 막을 수 있는 여러 종류의 생물반응기 시스템들이 개발되었다. 이러한 생물반응기 시스템의 개발과 우수한 분해능을 가지는 균주 선별 및 개량 등을 통해 실제 현장에 적용시킬 수 있는 생물학적 TCE 처리 기술의 실용화가 가능할 것이다.

• KSBB Journal
• /
• 제31권4호
• /
• pp.284-290
• /
• 2016

In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

• Asian-Australasian Journal of Animal Sciences
• /
• 제23권6호
• /
• pp.806-813
• /
• 2010

Animal bioreactors have been regarded as alternative tools for the production of limited human therapeutic proteins. The mammary glands of cattle are optimal tissues to produce therapeutic proteins that cannot be produced in large amounts in traditional systems based on microorganisms and eukaryotic cells. In this study, two knock-in vectors, pBCTPOKI-6 and pBCTPOKI-10, which target the hTPO gene on the bovine beta-casein locus, were designed to develop cloned transgenic cattle. The pBCTPOKI-6 and pBCTPOKI-10 vectors expressed hTPO protein in culture medium at a concentration of 774 pg/ml and 1,867 pg/ml, respectively. Successfully, two targeted cell clones were obtained from the bovine fibroblasts transfected with the pBCTPOKI-6 vector. Cloned embryos reconstructed with the targeted nuclei showed a lower in vitro developmental competence than those with the wild-type nuclei. After transfer of the cloned embryos into recipients, 7 pregnancies were detected at 40 to 60 days of gestation, but failed to develop to term. The results are the first trial for targeting of a human gene on the bovine milk protein gene locus, providing the potential for a large-scale production of therapeutic proteins in the animal bioreactor system.

• Journal of Microbiology and Biotechnology
• /
• 제20권3호
• /
• pp.447-459
• /
• 2010

Bacterial diversity of two distinct wastewater treatment systems, conventional activated sludge (CAS) and membrane bioreactor (MBR), of petroleum refineries were investigated through 16S rRNA gene libraries. Sequencing and phylogenetic analysis showed that the bacterial community composition of sludge samples was distinct between the two wastewater treatment systems. MBR clones belonged predominantly to Class Betaproteobacteria, represented mainly by genera Thiobacillus and Thauera, whereas CAS clones were mostly related to Class Alphaproteobacteria, represented by uncultured bacteria related to Order Parvularculales. Richness estimators ACE and Chao revealed that the diversity observed in both libraries at the species level is an underestimate of the total bacterial diversity present in the environment and further sampling would yield an increased observed diversity. Shannon and Simpson diversity indices were different between the libraries and revealed greater bacterial diversity for the MBR library, considering an evolutionary distance of 0.03. LIBSHUFF analyses revealed that MBR and CAS communities were significantly different at the 95% confidence level ($P{\leq}0.05$) for distances $0{\leq}D{\leq}0.20$. This work described, qualitatively and quantitatively, the structure of bacterial communities in industrial-scale MBR and CAS processes of the wastewater treatment system from petroleum refineries and demonstrated clearly differentiated communities responsible for the stable performance of wastewater treatment plants.

• Journal of Microbiology and Biotechnology
• /
• 제30권5호
• /
• pp.762-769
• /
• 2020

Vitamin K2 (menaquinone) is an essential vitamin existing in the daily diet, and menaquinone-7 (MK-7) is an important form of it. In a recent work, we engineered the synthesis modules of MK-7 in Bacillus subtilis, and the strain BS20 could produce 360 mg/l MK-7 in shake flasks, while the methylerythritol phosphate (MEP) pathway, which provides the precursor isopentenyl diphosphate for MK-7 synthesis, was not engineered. In this study, we overexpressed five genes of the MEP pathway in BS20 and finally obtained a strain (BS20DFHG) with MK-7 titer of 415 mg/l in shake flasks. Next, we optimized the fermentation process parameters (initial pH, temperature and aeration) in an 8-unit parallel bioreactor system consisting of 300-ml glass vessels. Based on this, we scaled up the MK-7 production by the strain BS20DFHG in a 50-l bioreactor, and the highest MK-7 titer reached 242 mg/l. Here, we show that the engineered strain BS20DFHG may be used for the industrial production of MK-7 in the future.

• KSBB Journal
• /
• 제25권5호
• /
• pp.471-477
• /
• 2010

In order to improve bio-ethanol productivity by various cultivation methods in this paper, the culture modes using food wastes, such as batch culture, high-cell-density fermentation, SSF (simultaneous saccharification and fermentation) by fill & draw, continuous culture by fill & draw were performed and their productivities were compared. SSFs by fill & draw were performed by continuous decompression using 1 L evaporator system, and by 10 L bioreactor without decompression. In addition, the continuous cultures by fill & draw mode using SFW (saccharafied food wastes) medium were performed by changes of 40% culture broth with intervals of 12 h (0.03 $h^{-1}$), 6 h (0.07 $h^{-1}$), 3 h (0.13 $h^{-1}$). Consequently, productivities of bio-ethanol were 2.52 g/L-h and 1.30 g/L-h in batch culture and high- cell-density fermentation, respectively. The productivities of SSF by fill & draw showed 2.24 g/L-h and 2.03 g/L-h in continuous decompression with 1 L evaporator and 10 L bioreactor without decompression, respectively. Also, the productivities in continuous culture by fill & draw modes showed 2.02 g/L-h, 4.07 g/L-h and 6.25 g/L-h by medium change with intervals of 12 h, 6 h, and 3 h, respectively. In conclusion, the highest ethanol productivity was obtained in the continuous culture mode by fill & draw with dilution rate of 0.13 $h^{-1}$.

• 한국항해항만학회:학술대회논문집
• /
• 한국항해항만학회 2010년도 춘계학술대회
• /
• pp.395-396
• /
• 2010

본 연구에서는 막결합형 연속회분석공정(MSBR)을 이용한 크루즈선 오 폐수처리장치 개발을 위하여 기초 실험을 수행하였다. MSBR 공정을 평가하기 위하여 SBR, MBR, MSBR system을 Lab scale로 제작하여 처리 효율과 크루즈선이라는 특수환경과의 접목성을 비교 운영하였으며, 검토 결과 MSBR 공정은 처리효율과 장치 운영면에서 선박이라는 특수한 환경에 가장 적합한 공정으로 평가되었다. MSBR의 system의 BOD 제거 효율은 99%, COD 제거 효율은 98%, SS 제거 효율은 99%로 나타내었으며 유기물, 영양염류, 병원성 미생물 처리에 있어 모두 안정적인 효율을 나타내며 IMO의 규제 기준을 모두 만족하였다.

• Journal of Microbiology and Biotechnology
• /
• 제17권1호
• /
• pp.116-122
• /
• 2007

An efficient enzyme system for the synthesis of L-tyrosine was developed using a fed-batch reactor with continuous feeding of phenol, pyruvate, and ammonia. A thermo- and chemostable tyrosine phenol-lyase from Symbiobacterium toebii was employed as the biocatalyst in this work. The enzyme was produced using a constitutive expression system in Escherichia coli BL21, and prepared as a soluble extract by rapid clarification, involving treatment with 40% methanol in the presence of excess ammonium chloride. The stability of the enzyme was maintained for at least 18 h under the synthesis conditions, including 75 mM phenol at pH 8.5 and $40^{\circ}C$. The fed-batch system (working volume, 0.51) containing 1.0 kU of the enzyme preparation was continuously fed with two substrate preparations: one containing 2.2 M phenol and 2.4 M sodium pyruvate, and the other containing 0.4 mM pyridoxal-5-phosphate and 4M ammonium chloride (pH 8.5). The system produced 130g/I of L-tyrosine within 30h, mostly as precipitated particles, upon continuous feeding of the substrates for 22 h. The maximum conversion yield of L-tyrosine was 94% on the basis of the supplied phenol.

• 청정기술
• /
• 제12권2호
• /
• pp.67-77
• /
• 2006

본 연구에서 소규모 하수고도처리를 위한 이중슬러지(Dual sludge) $KNR^{(R)}$ (Kwon's nutrient removal) 시스템이 개발되었다. $KNR^{(R)}$ 시스템은 부유성장식 탈질미생물과 부착성장식 질산화미생물을 분리시킨 이중슬러지 공정으로 최초침전조, 혐기조, 무산소조, 농축조의 복합기능을 수행하는 UMBR (Upflow multi-layer bioreactor)과 펠렛형 담체가 충진된 호기성 담체조로 구성되어 있다. 소규모 하수처리시 본 개발공정의 안정성과 처리성능을 평가하기 위해 처리용량 $50m^3/d$ 규모의 파일럿 플랜트를 고도처리 공정으로 개선공사 중인 처리용량 $50m^3/d$ 규모의 실제 소규모 마을하수처리장에 적용하였다. UMBR과 담체조의 체류시간은 각각 4.7 h와 7.2h이었으며, 반응조 수온은 $18.1{\sim}28.1^{\circ}C$이었다. 유입 하수량과 유입수의 BOD/N의 변동폭이 컸음에도 불구하고 파일럿 플랜트는 안정된 처리성능을 보였다. 전체 실험기간 중 처리수의 $COD_{cr}$, $COD_{Mn}$, $BOD_5$, TN, TP의 평균 농도는 11.0 mg/L, 8.8 mg/L, 4.2 mg/L, 3.5 mg/L, 9.8 mg/L, 0.87/0.17 mg/L (poly aluminium chloride(PAC) 투입/미투입)이었으며, 제거율을 각각 95.3%, 87.6%, 96.3%, 96.5%, 68.2%, 55.4/90.3% 이었다. 잉여슬러지 발생량은 $A_2O$와 Bardenpho 등과 같은 단일슬러지를 이용하는 고도처리공정과 비교시 약 1.9~3.8배 낮은 $0.026kg-DS/m^3$ and 0.220 kg-DS/kg-BOD로 나타났다.

• 대한환경공학회지
• /
• 제22권6호
• /
• pp.1149-1158
• /
• 2000

염색폐수를 생물학적으로 처리하여 방류하는 연속으로 연결되어있는 6개의 단으로 구성된 폐수처리시설에서 폐수 유동을 관찰하고 처리공정에 영향을 줄 수 있는 요인들을 찾기 위해 동위원소를 이용한 추적자 실험을 수행하였다. 추적자로 약 20mCi의 $^{131}I$를 첫 번째 단의 폐수 유입부에 투입하고 각 단에 방사선 검출기를 설치하여 시간에 따른 추적자 농도변화를 측정하였다. 측정된 데이터를 Perfect Mixers in Series Model을 이용한 분석을 통하여 평균체재시간과 시스템에서의 유동 특성 파라미터들을 계산하였다. 시스템의 평균체재시간은 설계치인 22.3시간의 76%인 17시간으로 계산되었다. 시스템을 구성하고 있는 6개의 단 중에서 제1단은 완전혼합기(perfect mixer)로서의 특성을 보이지 않았으나, 나머지 5개의 단은 완전혼합기와 같이 작동하였다. 제1단의 출력은 상호 교환이 일어나는 탱크를 가진 완전혼합기 모델로 모사한 출력과 좋은 일치를 보였다. 이 교환되는 탱크의 크기는 전체의 약 4분의 1인 것으로 분석되었다. 체재시간분포 실험의 결과는 유입 폐수의 방사선 전처리를 위해 설치된 전자선가속기의 가동조건 변경 후 그 영향에 대한 연구를 위한 적정 시료채취시간 산정에 사용되었다.