• Korean Journal of Microbiology
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• v.5 no.2
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• pp.61-68
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• 1967

In the process of the incorporation of orthophsphate into protein and other cell constituents, the role of inorganic polyphosphate and RNA-polyphosphate complex and the correlation between them were pursued by analyzing the contents of $^{32}P$ and total P in various fractions of Chlorella cells, which had been uniformly labeled with $^{32}P$ before the inoculation in a normal "cold" medium or P-free medium during the culture. The effects of ionizing radiation and various micronutritional-element deficiencies on the phosphate incorporation into, and biosynthesis of, protein and other introgenus compounds in the cells were also observed. When the uniformly $^{32}P$-labeled algae were grown in a normal "cold" medium the contents of $^{32}$ P in the fractions of protein, DNA and RNA-polyphosphate complex increased, but those in the fraction of acid-insoluble polyphosphate decreased. On the other hand, amount of $^{32}P$in the fraction of RNA was almost unchanged in spite of rapid increase of the total P. In the growing period of $^{32}P$-labeled algae in a P-free medium, amounts of $^{32}P$ in the fractions of DNA, protein and lipid increased, while those in the fractions of RNA-polyphosphate and inorganic polyphosphates decreased. When the algal cells were irradiated with about 70, 000r of gamma-rays before the inoculation in the medium, amounts of phosphate in the fractions of DNA, RNA, nucleotides and protein decreased during the culture, compared with those of the control. However, the phosphate content in the fraction of acid-insoluble polyphosphate of the irradiated cells increased than those of the control. In the growing period of the algae in a Mo-free, medium, amounts of acid-soluble total phosphate and nucleotides of the cells increased, while the amounts of residual protein and RNA decresed compared with those of the normal cells. Amounts of alkali-labile protein and phospholipid of the cells grown in a B-free medium decreased, whereas amount of phosphate in acid-soluble fraction increased compared with the control. In general, the contents of protein and RNA in each microelement deficient cells decreased more or less, compared with those in the normal cells.in the normal cells.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.274-279
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• 2015

Receptor activator of nuclear factor-kappa B ligand (RANKL) is a critical factor in osteoclastogenesis. It makes osteoclasts differentiate and multinucleate in bone remodeling. In the present study, RANKL was expressed as a soluble maltose binding protein (MBP)-fusion protein using the Escherichia coli maltose binding domain tag system (pMAL) expression vector system. The host cell E. coli DH5α was cultured and induced by isopropyl β-_D-1-thiogalactopyranoside for rRANKL expression. Cells were disrupted by sonication to collect soluble MBP-fused rRANKL. The MBP-fusion rRANKL was purified with MBP Trap affinity chromatography and treated with Tobacco Etch Virus nuclear inclusion endopeptidase (TEV protease) to remove the MBP fusion protein. Dialysis was then carried out to remove binding maltose from the cleaved rRANKL solution. The cleaved rRANKL was purified with a second MBP Trap affinity chromatography to separate unsevered MBP-fusion rRANKL and cleaved MBP fusion protein. The purified rRANKL was shown to have biological activity by performing in vitro cell tests. In conclusion, biologically active rRANKL was successfully purified by a simple two-step chromatography purification process with one column.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.5
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• pp.377-382
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• 1993

Effects of leaf and pod removal on changes in dry weight and on the contents of soluble sugar, starch, protein and oil in leaves and seeds of soybean [Glycine max(L.) Merr.] cultivar ‘Hwangkeumkong’ were measured at the research farm of Korea University in 1992. The upper 40% and lower 60% of leaves and pods were subjected to treatments at the growth stage of beginning pod(R3). Upper leaf-lower pod removal showed the highest leaf and the lowest seed dry weights. Soluble sugar content was no different among treatments in leaves and seeds. The highest starch content was found in leaves of upper leaf-lower pod removal. Protein content was higher in lower leaves than upper leaves and the lowest in seeds of lower leaf-upper pod removal which had the highest oil content in leaves and seeds. These results apparently indicated that photoassimilates were mobilized from upper leaves to lower seeds, and protein sources were moved from lower to upper parts but weak in remobilization from leaves for the long distance translocation during the reproductive growth period.

• Journal of The Korean Society of Grassland and Forage Science
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• v.21 no.2
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• pp.89-96
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• 2001

Carbohydrate and soluble protein reserves and regrowth characteristics in response to cutting height were investigated over four regrowth cycles of turf-type perennial ryegrass(Lolium perenne L. cv. preludeII). When the plants were at the full-vegetative stage (twelve weeks-old), three sequential defoliations at 3, 6 and 9 cm above the root base were imposed at 2-week intervals. Shoot dry weight in all three treatments continuously decreased with progressing regrowth cycle and the decreasing rate was higher as cutting height was lowered. TNC (total non-structural carbohydrate) in stubble at the end of the fourth regrowth cycle in 3, 6 and 9 cm cutting height decreased by 98%, 82% and 27%, respectively, comparing the initial content. TNC in roots also largely decreased with similar pattern in response to cutting height, whereas the absolute amount was much less compared to stubble. Soluble protein in stubble in 3, 6 and 9 cm cutting height decreased by 98%, 82% and 57%, respectively, at the end of fourth regrowth. A significant correlations between TNC (r=0.906) or protein (r=0.879) at the fourth defoliation and dry weight of regrowing shoots at the end of fourth regrowth were observed. these results indicated that cutting height closely influences the levels of organic reserves available for new growth, and that the levels of reserves might provide a useful tool as a determinant for regrowth dynamics.

• Molecules and Cells
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• v.26 no.1
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• pp.34-40
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• 2008

To express an increased level of recombinant Mefp1 (marine mussel adhesive protein) in soluble form, we constructed expression vectors encoding truncated OmpA signal peptide-Mefp1 fusion proteins. OmpA signal peptide (OmpASP) is the 21 residue peptide fragment of the 23 residue OmpA signal sequence cleavable by signal peptidase I. We successfully produced increased levels of soluble recombinant Mefp1 (rMefp1) with various deletions of OmpASP, and found that the increased expression was caused by the increased pI of the N-terminus of the fusion proteins (${\geq}10.55$). All the OmpA signal peptide segments of 3-21 amino acids in length had the same pI value (10.55). Our results suggest that the pI value of the truncated OmpASP ($OmpASP_{tr}$) play an important role in directional signaling for the fusion protein, but we found no evidence for the presence of a secretion enhancer in OmpASP. For practical applications, we increased the expression of soluble rMefp1 with $OmpASP_{tr}$ peptides as directional signals, and obtained rMefp1 with the native amino terminus (nN-rMefp1) using an $OmpASP_{tr}$ Xa leader sequence that contains the recognition site for Xa protease.

• The Korean Journal of Mycology
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• v.8 no.3
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• pp.159-166
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• 1980

Aspergillus niger van Tieghem was cultured by the method of submerged and synchronized culture for the study of differentiation. Acid-phenol soluble cell proteins of the fungus were extracted from four stages during development. Those acid-phenol soluble proteins were separated by polyacrylamide gel electrophoresis to determine the protein patterns. A new protein band was observed from the pre-sporulation body, color density of the stained protein bands in four tubes differed according to the differentiation stages. The number of protein bands in 10% gels varied from 18 to 16, 17, and 19 according to the course of spore germination stage, conidiophore stage, phialide maturation stage and sporulation stage.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.11a
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• pp.143-145
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• 1994

Chimeric fusion proteins involving IgG have proven valuable in studying protein-protein interactions and may possess therapeutic applications as well. For example, three receptor subtypes for the natriuretic peptides, when fused to the Fc portion of human IgG ${\gamma}$ chain, were quantitatively and qualitatively indistinguishable from the native receptor, thus allowing detailed structure-function studies of the receptor. In an attempt to block human immunodeficiency virus infectivity with soluble derivatives of CD4, a CD4/IgG Fc chimeric molecule was shown to increase the plasma half life of soluble CD4 and possessed the added advantage of IgG Fc-mediated placental transfer. In the case of the KGFR, this approach provided a framework for dissection of its ligand binding domains and made it possible to demonstrate that high affinity binding sites for two ligands, aFGF and KGF, reside within different receptor Ig-like domains. Chimeric molecules fused to immunoglobulins would have the advantages of secretion from transfected cells as well as detection and purification from medium utilizing Staphylococcus aureus Protein A. In addition, where highly related receptors make their discrimination very hard due to the difficulties in generating specific immunochemical probes, IgG fusion protein with tailor-made specificities confers particular advantages to elucidate patterns of receptor distribution and expression. The approach described here may have general applications in defining ligand-receptor interactions as well as searching for specific agonists and antagonists of receptor function.

• Preventive Nutrition and Food Science
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• v.20 no.4
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• pp.276-283
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• 2015

Hydrocolloids have many applications in foods including their use in dysphagia diets. We aimed to evaluate whether hydrocolloids in foods affect the digestibility of starch and protein, and their effects on antioxidant capacity. The thickening hydrocolloids: locust bean gum and carboxymethyl cellulose, and the gel-forming agents: agar agar, konjacglucomannan, and Hot & Soft Plus were blended with corn starch and soy protein, skim milk, or grape juice and were examined for their in vitro-digestability by comparing the reducing sugar and trichloroacetic acid (TCA)-soluble peptide, for antioxidant capacity by total polyphenol contents and the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity. The hydrocolloids resulted in a decrease in starch digestibility with the gel-forming agents. Hydrocolloids diminished TCA-soluble peptides in skim milk compared to soy protein with the exception of locust bean gum and decreased free radical scavenging capacities and total phenolic contents in grape juice. Our findings may provide evidence for the use of hydrocolloids for people at risk of nutritional deficiencies such as dysphagia patients.

• Journal of Animal Science and Technology
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• v.58 no.9
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• pp.33.1-33.7
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• 2016

Background: The continuing growth of the ethanol industry has generated large amounts of various distillers grains co-products. These are characterized by a wide variation in chemical composition and ruminal degradability. Therefore, their precise formulation in the ruminant diet requires the systematic evaluation of their degradation profiles in the rumen. Methods: Three distillers grains plus soluble co-products (DDGS) namely, corn DDGS, high-protein corn DDGS (HP-DDGS), and wheat DDGS, were subjected to an in situ trial to determine the degradation kinetics of the dry matter (DM) and crude protein (CP). Soybean meal (SBM), a feed with highly degradable protein in the rumen, was included as the fourth feed. The four feeds were incubated in duplicate at each time point in the rumen of three ruminally cannulated Hanwoo cattle for 1, 2, 4, 6, 8, 12, 24, and 48 h. Results: Wheat DDGS had the highest filterable and soluble A fraction of its DM (37.2 %), but the lowest degradable B (49.5 %; P < 0.001) and an undegradable C fraction (13.3 %; P < 0.001). The filterable and soluble A fraction of CP was greatest with wheat DDGS, intermediate with corn DDGS, and lowest with HP-DDGS and SBM; however, the undegradable C fraction of CP was the greatest with HP-DDGS (41.2 %), intermediate with corn DDGS (2.7 %), and lowest with wheat DDGS and SMB (average 4.3 %). The degradation rate of degradable B fraction ($%\;h^{-1}$) was ranked from highest to lowest as follows for 1) DM: SBM (13.3), wheat DDGS (9.1), and corn DDGS and HP-DDGS (average 5.2); 2) CP: SBM (17.6), wheat DDGS (11.6), and corn DDGS and HP-DDGS (average 4.4). The in situ effective degradability of CP, assuming a passage rate of $0.06h^{-1}$, was the highest (P < 0.001) for SBM (73.9 %) and wheat DDGS (71.2 %), intermediate for corn DDGS (42.5 %), and the lowest for HP-DDGS (28.6 %), which suggests that corn DDGS and HP-DDGS are a good source of undegraded intake protein for ruminants. Conclusions: This study provided a comparative estimate of ruminal DM and CP degradation characteristics for three DDGS co-products and SBM, which might be useful for their inclusion in the diet according to the ruminally undegraded to degraded intake protein ratio.

• Animal cells and systems
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• v.15 no.1
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• pp.29-36
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• 2011

Insect lysozymes are basic, cationic proteins synthesized in fat body and hemocytes in response to bacterial infections and depolymerize the bacterial cell wall. The c-type lysozyme of the insect Spodoptera litura (SLLyz) is a single polypeptide chain of 121 residues with four disulfide bridges and 17 rare codons and is approximately 15 kDa. The full-length SLLyz cDNA is 1039 bp long with a poly(A) tail, and contains an open reading frame of 426 bp long (including the termination codon), flanked by a 54 bp long 5' UTR and a 559 bp long 3' UTR. As a host for the production of high-level recombinant proteins, E. coli is used most commonly because of its low cost and short generation time. However, the soluble expression of heterologous proteins in E. coli is not trivial, especially for disulfide-bonded proteins. In order to prevent inclusion body formation, GST was selected as a fusion partner to enhance the solubility of recombinant protein, and fused to the amplified products encoding mature SLLyz. The expression vector pGEX-4T-1/rSLLyz was then transformed into E. coli BL21(DE3)pLysS for soluble expression of rSLLyz, and the soluble fusion protein was purified successfully. Inhibition zone assay demonstrated that rSLLyz showed antibacterial activity against B. megaterium. These results demonstrate that the GST fusion expression system in E. coli described in this study is efficient and inexpensive in producing a disulfide-bonded rSLLyz in soluble, active form, and suggest that the insect lysozyme is an interesting system for future structural and functional studies.