• Journal of Plant Biology
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• 제38권1호
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• pp.1-10
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• 1995

Photoinhibition and its recovery of spectrally adapted Anacystis nidulans were studied. Phycocyanin and Chl content and phycocyanin/Chl ratio were increased in cells grown under blue-green light compared with those grown in white light. Photosynthetic activities of white light and blue-green light grown cells were reduced by 50% after 15 min and 10 min of photoinhibitory light treatment (1.2 mmol·m-2s-1), respectively, largely due to the decline of PSII activities. However, their activities were recovered fully after 30 min incubation under weak light. Treatment of rifampicin and chloramphenicol magnified the photoinhibitory effects and suppressed the recovery with disappearance of susceptibility to photoinhibition and delayed the recovery process, indicating no significant differences in phosphorylation, dephosphorylation and protease activity between two cells. Therefore, it is suggested that the increased sensitivity of blue-green adapted cells might be attributed to the decline of protein synthesis, and phosphorylation-dephosphorylation of protein and protease activity might be involved in the recovery process.

• 생명과학회지
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• 제15권1호
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• pp.9-14
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• 2005

난황 단백질의 효소에 의한 가수분해에서 에탄올의 농도가 증가할수록 고형물과 단백질의 회수가 증가함을 알 수 있었다. 이것은 유화층을 에탄올의 증가가 감소시켰기 때문이다. 그러나 가수분해물에서 단백질 함량이나 sialic acid의 함량은 에탄을 농도와는 관계없이 일정하였다. 한외여과 후에 retentate에 대한 에탄올의 영향을 조사하였다. 마찬가지로 고형물의 회수는 에탄을 농도의 증가와 함께 증가하였다. 그리고 retentate에서 sialic acid의 함량은 대략 $2.5\%$정도로 일정하고 에탄올의 농도에 영향을 받지 않았다 이상의 결과로부터 난황 단백질의 효소 가수분해를 통해 sialic acid가 함유된 제품을 얻고자 할 때는 원료 난황단백질에 포함된 에탄올의 함량을 증가시킬수록 높은 수율의 제품을 얻을 수 있다. 본 실험에서는 원료 난황 단백질 중에서 $40\%$의 에탄을 함량까지는 제품 수율이 계속 증가하는 경향을 보였다. 난황 단백질 가수분해물의 한외여과에서 농축단계에서는 막 모듈의 MWCO의 차이에도 불구하고 retentate에서 총 고형물의 회수율은 비슷함을 나타내었으며, 투석에서 회수율은 MWCO가 작을수록 높아지지는 않았다. 제품에서의 sialic acid의 함량은 사용한 모듈에서 약 $2.0\%$를 나타내었다. 이것은 단백질 가수분해물에 비해 5배정도 상승한 결과이다. 본 연구에서 사용된 막 모듈 가운데서 Amicon 모듈이 제품의 회수율과 함량면에서 가장 우수한 특성을 보였다. 결론적으로 난황 단백질의 protease에 의한 가수분해에서 한외여과에 의해 순도를 높일 때 MWCO, 막 모듈의 type 그리고 운전조건 등을 잘 고려해 줄 때 최적의 조업조건을 얻을 수 있었다.

• 멤브레인
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• 제12권3호
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• pp.165-170
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• 2002

씻어나온 쌀(무세미) 생산공정 중 발생하는 쌀뜨물 중 함유된 유효성분, 특히 단백질 회수를 위한 분리막 공정에 관하여 검토하였다. 먼저 dead-end형 Amicon 여과셀을 이용하여 단백질 농축에 적절한 분리막을 선정하였으며 이를 토대로 중공사형 한외여과 모듈 또는 가정용 정수기에 사용되는 나권형 나노여과 및 역삼투 모듈을 이용한 투과실험을 행하였다. 그 결과, 분획분자량이 10,000 dalton인 한외여과막은 쌀뜨물내 유효성분 내지는 단백q질을 농축하기에 적절하지 않았다. 그러나 역삼투 또는 나노여과 모듈에 9% 가량의 단백질이 함유된 원료용액을 250%까지 농축할 경우, 역삼투 모듈 농축액중에 단백질의 농도는 22%로서 약 2.4배가 농축되었으며 나노여과 모듈의 경우는 약 2배까지 단백질을 농축할 수 있었다.

• 한국축산식품학회지
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• 제35권1호
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• pp.50-57
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• 2015

Fish sarcoplasmic protein (SP) is currently dumped as waste from surimi industry and its recovery by practical method for being the non-meat ingredient in meat industry would be a strategy to utilize effectively the fish resource. This study was aimed to apply pH treatment for fish SP recovery and evaluated its effect on pork myofibrillar protein (MP) gel. The pH values of fish SP were changed to 3 and 12, and neutralized to pH 7 before lyophilizing the precipitated protein after centrifugation. Acid-treated fish SP (AFSP) showed about 4-fold higher recovery yield than that of alkaline-treated SP and water absorption capacity was also about 1.2-fold greater. Because of the high recovery yield and water absorption capacity, AFSP was selected to incorporate into MP with/without microbial transglutaminase (MTG). The effects of AFSP and MTG on the physicochemical and rheological characteristics of MP and MP gel were evaluated. MTG induced an increase shear stress of the MP mixture and increase the breaking force of MP gels. MP gel lightness was decreased by adding AFSP. MP gel with MTG showed higher cooking loss than that without MTG. A reduction of cooking loss was observed when the AFSP was added along with MTG, where the insoluble particles were found. Therefore, AFSP could be contributed as a water holding agent in meat protein gel.

• 한국식품저장유통학회지
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• 제22권2호
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• pp.256-260
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• 2015

본 연구에서는 알칼리 사용에 따른 갈변을 최소화하고 영양적으로 우수한 천마 단백질을 보다 효율적으로 추출하기 위하여 용출 및 침전 pH에 따른 단백질의 갈변도와 함량을 측정함으로써 탈지 천마 단백질 알칼리 추출의 최적 pH 조건을 설정하고자 하였다. 천마의 수분은 약 15%로 측정되었으며, 대부분 탄수화물로 구성되어 있었고, 단백질의 함량이 높은 것으로 확인되었다. 이러한 단백질을 다양한 pH에서 용출시킨 결과 pH가 증가함에 따라 용출된 단백질의 양이 증가하였으며, 갈변도 또한 증가함을 보였다. 침전 pH에 따른 단백질 함량은 pH 4에서 침전된 pellet이 가장 많은 함량을 나타내었으며, 상등액의 단백질 함량 또한 대부분 pH 4로 침전시킨 경우 가장 적은 것으로 확인되었다. 뿐만 아니라 단백질의 회수율도 침전 pH가 4일 때 가장 높은 값을 나타내었다. 따라서 갈변도와 단백질 함량을 고려한 탈지 천마 단백질 추출을 위한 최적조건은 용출 pH 9와 침전 pH 4로 결정되었다.

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

• 한국응용과학기술학회지
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• 제24권4호
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• pp.347-353
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• 2007

Fleshing scrap is a kind of wastes produced during leather making process and used in the test of manufacturing biodiesel. The early step of manufacturing biodiesel is fat recovery from fleshing scrap. Hence, we investigated the influence of the way of fat recovery on the fatty acid composition. We used three different recovery ways, that is chemical method by protein decomposition with acid/fat recovering, physical method by protein denaturalization with heat and vacuum/fat pressing, and biodiesel method by protein decomposition/fat recovering. The biological method yielded the best results in terms of appearance transparency. It was most effective to lower acid value. Also the recovered fat by biological method would be favorable methyl-ester reaction raw material for biodiesel because it contains more than 5% of oleic acid among unsaturated fatty acid.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권6호
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• pp.419-425
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• 2001

A dense pellicular solid matrix has been fabricated by coating 4% agarose gel on to dense zironia-silica(ZS) spheres by watr-in-oil emulsification . The agarose evenly laminated the ZS bead to a depth of 30㎛, and the resultin gpellicular assembly was characterised by densities up to 2.39g/mL and a mean particle dimeter of 136 ㎛. In comparative fluidisation tests, the pellicular solid phase exhibited a two-fold greater flow velocity than commercial benchmark ad-sorbents necessary to achieve common values of bed expansion. Furthermore, the perlicular parti-cles were characterised by improved qualities of chromatographic behaviour, particularly with re-spect to a three-fold increase in the apparent effective diffusivity of lysozyme within a pellicular assembly modified with Cibacron Blue 3GA. The properties of rapid protein adsorption/desorp-tion were attributed to the physical design and pellicular deployment of the reactive surface in the solid phase. When combined with enhanced feedstock throughput, such practical advantages recommend the pellicular assembly as a base matrix for the selective recovery of protein products from complex, particulate feedstocks(whole fermentation broths, cell disruptates and biological extracts).

• Journal of Microbiology and Biotechnology
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• 제2권2호
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• pp.135-140
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• 1992

The partitioning of recombinant human interleukin-2(rhII-2) in PEG 8000-dextran 38800 aqueous two-phase system has been investigated using three different sources of rhIL-2. In the case of pure rhIL-2, the solubility in a PEG-dextran two-phase system was low and most of rhIL-2 was partitioned into the bottom phase. For the recovery of rhIL-2 from insoluble protein aggregates, the inclusion bodies of recombinant E. coli were solubilized by the treatment with sodium dodecyl sulfate (SDS). The addition of SDS significantly enhanced not only the solubility of rhIL-2 but also the partitioning of rhIL-2 to the top phase. When the ratio of SDS to rhIL-2 was 2.0, the partition coefficient(K) and the recovery yield(Y) at the top phase were 4.5 and 88%, respectively, at pH 6.8. In order to reduce the recovery steps further, SDS was directly added to the intact recombinant E. coli cells and then partitioned into the PEG/dextran aqueous two-phase system. The observed partition coefficient ($K{\cong{3.0$) and recovery yield ($Y{\geq}80%$ )of this method were comparable to the rhIL-2 recovery from insoluble protein aggregates. The results obtained in this work indicate that PEG-dextran two-phase partitioning might provide a simple way for the recovery and partial purification of recombinant proteins which are produced as inclusion bodies.

• 산업식품공학
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• 제15권2호
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• pp.143-147
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• 2011

알칼리를 이용하여 탈지미강에서 단백질을 추출하기 위해 용출 pH(8, 9, 10, 11, 12) 5개 구간과 등전점 침전 pH(2, 4, 6) 3개 구간을 설정하고 각 pH 구간에서의 회수된 단백질 함량과 갈변, 전기영동 패턴 및 최종 수용성 단백질의 회수율을 비교 확인하였다. 용출 pH가 증가할수록 수용성 단백질의 회수율은 증가하였으나 동시에 갈변현상도 증가하였다. 등전점 침전 pH는 4에서 가장 많은 단백질을 회수할 수 있었다. SDS-PAGE 결과, 사용한 pH에 따라 추출된 단백질의 주요 패턴은 뚜렷한 차이가 없었으나, 높은 알칼리조건이 35 kDa 이하의 단백질 추출에 좀 더 용이하였다. 각 pH 구간별 용출 대비 수용성 단백질 회수율은 평균 31.5% 이었다. 용출 pH 11 및 침전 pH 4 구간에서 46.95%의 가장 높은 회수율을 나타냈으나 갈변이 급격히 증가하였다. 따라서 37.65%의 회수율을 나타낸 용출 pH 10 과 침전 pH 4 구간이 미강 단백질의 최적 추출조건임을 확인하였다.