• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.658-662
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• 2008

Effects of pH shock on the secretion system of S. coelicolor A3(2) have been investigated at a transcriptional level by using DNA microarrays. Actinorhodin secretion was observed to be highly enhanced when an acidic-pH shock was applied to surface grown cultures of S. coelicolor A3(2). In this culture, a gene of actVA-orf1 encoding a putative efflux pump or transporter protein for actinorhodin was strongly upregulated. A major number of efflux pumps for other metabolites and a major number of secretion proteins for protein secretion were also observed to be upregulated with pH shock. The secretion of actinorhodin was observed to be remarkably enhanced in liquid culture as well.

• Asian-Australasian Journal of Animal Sciences
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• 제25권8호
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• pp.1083-1088
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• 2012

The breeding value of marbling score in skeletal muscle is an important factor for evaluating beef quality. In the present study, we investigated proteins associated with the breeding value of the marbling score for bovine sirloin to select potential biomarkers to improve meat quality through comparative proteomic analysis. Proteins isolated from muscle were separated by two-dimensional gel electrophoresis. After analyzing images of the stained gel, seven protein spots for the high marbling score group were identified corresponding to changes in expression that were at least two-fold compared to the low marbling score group. Four spots with increased intensities in the high marbling score group were identified as phosphoglycerate kinase 1, triosephophate isomerase, acidic ribosomal phosphoprotein PO, and capping protein (actin filament) Z-line alpha 2. Spots with decreased intensities in the high marbling score group compared to the low score group were identified as 14-3-3 epsilon, carbonic anhydrase II, and myosin light chain 1. Expression of myosin light chain 1 and carbonic anhydrase 2 was confirmed by Western blotting. Taken together, these data could help improve the economic performance of cattle and provide useful information about the underlying the function of bovine skeletal muscle.

• Biomolecules & Therapeutics
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• 제21권1호
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• pp.1-9
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• 2013

Polyamines, putrescine, spermidine and spermine, are ubiquitous in living cells and are essential for eukaryotic cell growth. These polycations interact with negatively charged molecules such as DNA, RNA, acidic proteins and phospholipids and modulate various cellular functions including macromolecular synthesis. Dysregulation of the polyamine pathway leads to pathological conditions including cancer, inflammation, stroke, renal failure and diabetes. Increase in polyamines and polyamine synthesis enzymes is often associated with tumor growth, and urinary and plasma contents of polyamines and their metabolites have been investigated as diagnostic markers for cancers. Of these, diacetylated derivatives of spermidine and spermine are elevated in the urine of cancer patients and present potential markers for early detection. Enhanced catabolism of cellular polyamines by polyamine oxidases (PAO), spermine oxidase (SMO) or acetylpolyamine oxidase (AcPAO), increases cellular oxidative stress and generates hydrogen peroxide and a reactive toxic metabolite, acrolein, which covalently incorporates into lysine residues of cellular proteins. Levels of protein-conjuagated acrolein (PC-Acro) and polyamine oxidizing enzymes were increased in the locus of brain infarction and in plasma in a mouse model of stroke and also in the plasma of stroke patients. When the combined measurements of PC-Acro, interleukin 6 (IL-6), and C-reactive protein (CRP) were evaluated, even silent brain infarction (SBI) was detected with high sensitivity and specificity. Considering that there are no reliable biochemical markers for early stage of stroke, PC-Acro and PAOs present promising markers. Thus the polyamine metabolites in plasma or urine provide useful tools in early diagnosis of cancer and stroke.

• Bulletin of the Korean Chemical Society
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• 제31권12호
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• pp.3521-3524
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• 2010

MTH1880 is a hypothetical protein derived from Methanobacterium thermoautotrophicum, thermophilic methanogen. The solution structure determined by NMR spectroscopy showed that it has a novel $\alpha+\beta$-fold with a highly acidic ligand binding pocket. Since MTH1880 maintains its ultra-stable structural characteristics at both high temperature and pressure, it has been considered as an excellent model for studying protein folding. To initiate the structural and folding study of MTH1880 in proving its unusual stability, we performed the site directed mutagenesis and biochemical analysis of MTH1880 mutants. Data from circular dichroism and NMR spectroscopy suggest that the point mutations perturbed the structural stability of protein even though the secondary structure is retained. This study will provide the useful information in understanding the role of participating residues during folding-unfolding process and our result will be used in designing further folding experiments for hyper-thermopile proteins like MTH1880.

• Molecules and Cells
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• 제42권10호
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• pp.729-738
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• 2019

Autophagy is an important process for protein recycling. Oligomerization of p62/SQSTM1 is an essential step in this process and is achieved in two steps. Phox and Bem1p (PB1) domains can oligomerize through both basic and acidic surfaces in each molecule. The ZZ-type zinc finger (ZZ) domain binds to target proteins and promotes higher-oligomerization of p62. This mechanism is an important step in routing target proteins to the autophagosome. Here, we determined the crystal structure of the PB1 homo-dimer and modeled the p62 PB1 oligomers. These oligomer models were represented by a cylindrical helix and were compared with the previously determined electron microscopic map of a PB1 oligomer. To accurately compare, we mathematically calculated the lead length and radius of the helical oligomers. Our PB1 oligomer model fits the electron microscopy map and is both bendable and stretchable as a flexible helical filament.

• 한국축산식품학회지
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• 제37권2호
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• pp.162-167
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• 2017

Functional and nutritional soluble proteins can be recovered from surimi (and surimi-like material) processing wastewater, reducing environmental problems and the cost of an irresponsible waste disposal. Recovered proteins may be added back at a low percentage to surimi products. The aim of this work was to evaluate the effect of the addition of soluble recovered proteins (RP), obtained from mechanically separated chicken meat surimi-like material (MSCM-SLM) processing wastewater by acidic pH-shifting, on the composition and texture of RP-MSCM-SLM, with RP contents of 0, 10, 20 and 30% (w/w) in the mixture. For that, proximate composition and gel properties were evaluated. The fat content of the MSCM-SLM was significantly reduced to 11.98% and protein increased to 83.64% (dry basis) after three washing cycles. The addition of 30% RP in the MSCM-SLM significantly augmented the protein content to 93.45% and reduced fat content from to 2.78%. On the other hand, the addition of RP was responsible for a drastic decrease in texture parameters, reaching 252.36 g, 185.23 g.cm, and 6.97 N for breaking force, gel strength and cutting strength, respectively, when 30% of RP was included in the MSCM-SLM. It was concluded that the obtained intermediary product (RP-MSCM-SLM) is a good option to applications in processed meat products where high texture parameters are dispensable, e.g., emulsified inlaid frankfurter-type sausages, but high protein content and low fat content desired.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.280-287
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• 2015

Current influenza vaccines are produced in embryonated chicken eggs. However, egg-based vaccines have various problems. To address these problems, recombinant protein vaccines have been developed as new vaccine candidates. Unfortunately, recombinant proteins frequently encounter aggregation and low stability during their biogenesis. It has been previously demonstrated that recombinantly expressed proteins can be greatly stabilized with high solubility by fusing stabilizing peptide (SP) derived from the C-terminal acidic tail of human synuclein (ATS). To investigate whether SP fusion proteins can induce protective immunity in mice, we produced influenza HA and SP fusion protein using a baculovirus expression system. In in vitro tests, SP-fused recombinant HA1 (SP-rHA1) was shown to be more stable than recombinant HA1 (rHA1). Mice were immunized intramuscularly with baculovirus-expressed rHA1 protein or SP-rHA1 protein ($2{\mu}g/mouse$) formulated with aluminum hydroxide. Antibody responses were determined by ELISA and hemagglutination inhibition assay. We observed that SP-rHA1 immunization elicited HA-specific antibody responses that were comparable to rHA1 immunization. These results indicate that fusion of SP to rHA1 does not negatively affect the immunogenicity of the vaccine candidate. Therefore, it is possible to apply SP fusion technology to develop stable recombinant protein vaccines with high solubility.

• Applied Biological Chemistry
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• 제25권4호
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• pp.248-251
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• 1982

3가지 단백질과 CBG250과의 반응에 의해서 생성되는 복합체(複合體)의 분광학적(分光學的) 성질(性質)과 결합당량(結合當量)을 측정(測定)하였다. CBG250은 사용한 몇 가지 용매(溶媒)에 따라 최대흡광파장(最大吸光波長)$({\lambda}_m)$이 이동(移動)하였으며, ethanol용액 중 $({\lambda}_m=610nm)$에서 흡광계수(吸光係數)는 82.4, 몰 흡광계수(吸光係數)는 $70.4{\times}10^3$이었다. CBG250은 ethanol―인산(燐酸)―수용액(水溶液)에서 갈색$({\lambda}_m=465nm)$을 가지나 일단 단백질과 결합하면 청색(靑色)$({\lambda}_m=590nm)$으로 변환되었으며, 파장(波長) 590nm에서의 흡광도(吸光度)와 단백질 함량(含量) 사이에는 제한된 범위에서 비례관계를 나타냈다. 반응조건에서 단백질과 CBG250은 신속하게 복합체(複合體)를 형성하였다. 단백질의 CBG250에 대한 결합당량(結合當量)은 단백질의 종류(種類)와 함량(含量)에 따라 현저하게 변하였다. BSA, Cytochrome C, ${\gamma}-globulin$의 그것은 각각 110, 103, $88{\mu}g\;$CBG250/100{\mu}g$ protein이었다.

• KSBB Journal
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• 제14권6호
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• pp.677-681
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• 1999

계란의 난황으로부터 유래한 IgY의 분리와 정제과정이 흐름을 정리한 결과, 난황에서의 수용성 단백질의 분리단계가 단백질의 수율을 증가시키는데 가장 큰 영향을 미치는 것으로 판단되었다. 본 연구에서는 자연 침전방법을 통하여 실험하였는데 수율의 증가를 위해서는 원심분리와 같이 수용성 단백질의 양을 많이 얻을 수 있는 방법이 필요하고, 인지질이나 lipoprotein등의 성분의 제거효율을 높이기 위해서는 x-carrageenan등과 같은 natural gum성분의 첨가와 같은 방법이 필요하다. 그러므로 자연침전을 이용하여 수율의 증가측면과 비용의 절감의 효과를 얻을 수 있었다. 보다 효율적인 IgY의 분리를 위해서는 초기 단계에서 수율의 극대화와 불순성분 제거의 최대화가 동시에 요구되고 있다. 전처리 후의 시료를 이용하여 이온교환 크로마토그래피와 gel filtration chromatography를 통해 순도 90% 이상의 IgY를 얻을 수 있었다.

• 한국식품과학회지
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• 제25권4호
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• pp.360-365
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• 1993

Phytate 함량이 다른 두 분리 대두단백(HSPI; high-phytate soy protein isolate, LSPI; low-phytate soy protein isolate)을 제조하여 pH 및 phytate 첨가량이 그들의 용해도와 소화율에 미치는 영향을 알아보았고 분리 대두단백을 용해도에 따라 분별한 후 그들의 특성을 전기영동을 통하여 알아보았다. LSPI와 HSPI 모두 분자량이 $13.6{\sim}81.1kDa$에 이르는 단백질 성분으로 이루어져 있으며 산성 pH에서 $18.0{\sim}35.0kDa$의 단백질은 phytate와 잘 결합하지 않는 특성을 가졌다. LSPI를 용해도에 다라 분별하였을 때 gliadin을 구성하는 단백질은 pH2에서 phytate와 쉽게 결합하지 않았다. 대두 단백질은 pepsin 소화율은 phytate 첨가량이 증가할수록 크게 저하되었다.