• Korean Journal of Microbiology
• /
• v.28 no.4
• /
• pp.297-303
• /
• 1990

Secretion of Serratia marcescens nuclease by E. coli harboring pNUC4 was investigated. 29.2, 54.2 and 16.6% of total nuclease were observed in culture medium, periplasm, and cytoplasm of E. coli, respectively. To investigate the secretion mechanism of Serratia nuclease by E. coli, secretion kinetics of nuclease was examined in the presences of sodium azide, and energy metabolism inhibitor; procaine, an exoprotein processing inhibitor; and chloramphenicol, a protein synthesis inhibitor. In the presence of sodium azide, periplasmic unclease was gradually decreased and the extracellular nyclease was linearly increased according to the incubation time. Similar results were obtained in presences of procaine and chloramphenicol. From these results, we concluded that two transport processes are involved in nuclease secretion: secretion of nuclease through the inner membrane is occurred by an energy-dependent process and probably requiring precusor processing: secretion of nuclease through outer membrane does not require energy, de novo protein synthesis, and precursor processing.

• Journal of Life Science
• /
• v.11 no.2
• /
• pp.108-110
• /
• 2001

The E. coli expression system to overproduce a harmful protein, carboxypeptidase Taq was developed. Since expression plasmid pCK305N containing the colicin promoter already has the initiation codon on the restriction site, the initiation codon of the CPase Taq gene was removed. Expression plasmid pCP4-col includes the entire CPase Taq gene, which is directed by the colicin promoter. E. coli cells harboring pCP-col produced a high amount of the enzyme when they were cultured in the present of mitomycin C (0.4 ${\mu}g$/ml). An amount of purified enzyme produced by pCP4-col directed by the colicin promoter was 10.5 mg. This result indicated that the novel E. coli expression system controlled by the colicin promoter could produce almost twice amounts of CPase Taq than the conventional system controlled by the tart promoter.

• Food Science and Preservation
• /
• v.3 no.2
• /
• pp.187-193
• /
• 1996

To investigate the antimicrobial activities and effect of grapefruit seed extract(GFSE) on the physiological function of microorganism, antimicrobial activity, fatty acids of bacterial cell lipid and amino acids of bacterial cell protein were measured. The change of cell morphotype was observed by transmission electron microscope. GFSE was very stable on the wide range temperture and pH. The growth rate of E. coli and B. suvtilis were decreased above 40ppm GFSE There fore, minimum inhibitory concentration (MIC) of the E. coli and B. subtilis to GFSE were determined around 40ppm. In the change of fatty acids quantities, hexadecanoate was significantly decreased on the treatment compared with control in case of E. coli, whereas tridecanoate was not detected in case of B. subtilis. In the change of amino acids quantities, alanine, glutamic acid, glycine, lysine were decreased on the treatment compared with control in case of E. coli and B. subtilis Transmission electron microsgraphs(TEM) showed the microbial cells were destroyed by GFSE.

• The Korean Journal of Food And Nutrition
• /
• v.8 no.1
• /
• pp.37-42
• /
• 1995

The amino acid sequence of basic isozyme 55 of Horseradish Peroxidase (HRP E5) was determined by protein sequencing. HRP E5 consisted about 300 residues, and has a molecular weight of approximately 36,000 $\pm$ 500 dalton. The protein was rich In aspartic acid (14%), arginine(13%), and leucine(11%). The primary structure of HRP E5 was established by sequencing its tryptic (T1-T19) and lysylendopeptic (Al-A3) peptides. The sequence homology between HRP E5 and HRP C (neutral isozyme of horseradish peroxidase) is found to be more than 66%. The highest concentration of identical residues are found on residues 29~56, 90~123, and 155~173, but relatively low on 174~271.

• Animal Bioscience
• /
• v.35 no.2
• /
• pp.260-271
• /
• 2022

Objective: We aimed to investigate the effect of the differing amino acid (AA) release dynamics of two protein sources on the growth performance, nitrogen deposition, plasma biochemical parameters, and muscle synthesis and degradation of piglets when included in their diets at normal and low concentrations. Methods: Forty-eight piglets (Duroc×Landrace×Large White) with initial body weight of 7.45±0.58 kg were assigned to six groups and fed one of 6 diets. The 6 dietary treatments were arranged by 3×2 factorial with 3 protein sources and 2 dietary protein levels. They are NCAS (a normal protein content with casein), NBlend (a normal protein content with blend of casein and corn gluten meal), NCGM (a normal protein content with corn gluten meal), LCAS (a low protein content with casein), LBlend (a low protein content with blend of casein and corn gluten meal), LCGM (a low protein content with corn gluten meal). The release dynamics of AA in these diets were determined by in vitro digestion. The digestibility, utilization and biological value of nitrogen in piglets were determined by micro Kjeldahl method. Plasma insulin was measured by enzyme-linked immunosorbent assay kits. The protein expression of mediators of muscle synthesis and degradation was determined by western blotting. Results: Although the consumption of a low-protein diet supplemented with crystalline AA was associated with greater nitrogen digestion and utilization (p<0.05), the final body weight, growth performance, nitrogen deposition, and phosphorylation of ribosomal protein S6 kinase 1 and eIF4E binding protein 1 in the muscle of pigs in the low-protein diet-fed groups were lower than those of the normal-protein diet-fed groups (p<0.05) because of the absence of non-essential AA. Because of the more balanced release of AA, the casein (CAS) and Blend-fed groups showed superior growth performance, final body weight and nitrogen deposition, and lower expression of muscle ring finger 1 and muscle atrophy F-box than the CGM-fed groups (p<0.05). Conclusion: We conclude that the balanced release of AA from CAS containing diets and mixed diets could reduce muscle degradation, favor nitrogen retention, % intake and improve growth performance in pigs consuming either a normal- or low-protein diet.

• Molecules and Cells
• /
• v.23 no.1
• /
• pp.115-121
• /
• 2007

Root nodule formation is controlled by plant hormones such as auxin. Auxin-repressed protein (ARP) genes have been identified in various plant species but their functions are not clear. We have isolated a full-length cDNA clone (EuNOD-ARP1) showing high sequence homology to previously identified ARP genes from root nodules of Elaeagnus umbellata. Genomic Southern hybridization showed that there are at least four ARP-related genes in the genome of E. umbellata. The cDNA clone encodes a polypeptide of 120 amino acid residues with no signal peptide or organelle-targeting signals, indicating that it is a cytosolic protein. Its cytosolic location was confirmed using Arabidopsis protoplasts expressing a EuNOD-ARP1:smGFP fusion protein. Northern hybridization showed that EuNOD-ARP1 expression was higher in root nodules than in leaves or uninoculated roots. Unlike the ARP genes of strawberry and black locust, which are negatively regulated by exogenous auxin, EuNOD-ARP1 expression is induced by auxin in leaf tissue of E. umbellata. In situ hybridization revealed that EuNOD-ARP1 is mainly expressed in the fixation zone of root nodules.

• Korean Journal of Food Science and Technology
• /
• v.30 no.2
• /
• pp.372-378
• /
• 1998

Two types of formaldehyde-treated soy protein isolate (SPI) films, formaldehyde-incorporated and formaldehyde-adsorbed films, and control SPI films were prepared. Cross-linking effect of formaldehyde on selected film properties such as color, tensile strength (TS), elongation at break (E), water vapor permeability (WVP), and water solubility (WS) were determined. Physical properties of formaldehyde-incorporated films were not geneally different from those of control films, while almost all of those among formaldehyde-adsorbed films were significantly different. Through cross-linking development within formaldehyde-adsorbed films, WS decreased significantly (P<0.05) from 26.1% to 16.6%, and TS increased two times while E decreased two times compared with control films. This was caused by insolubilization and hardening of protein by cross-linking most likely attributed to the significant changes in properties of protein films reacted with formaldehyde.

• Journal of Microbiology and Biotechnology
• /
• v.8 no.6
• /
• pp.663-668
• /
• 1998

Several neurodegenerative diseases such as Huntington's disease, dentatorubralpallidoluysian atrophy, spinobulbar muscular atrophy, Machado-Joseph disease, and spinocerebellar ataxias type 1 are associated with the aggregation of expanded glutamine repeats within their proteins. Generally, in clinically affected individuals, the expansion of the polyglutamine sequences is beyond 40 residues. To address the length of polyglutamine that forms aggregation, we have constructed plasmids encoding glutathione S-transferase (GST) Machado-Joseph disease gene fusion proteins containing polyglutamine and investigated the formation of aggregates in E. coli. Surprisingly, even $(Gin)_8$, in the normal range as well as $(Gin)_{65}$ in the pathogenic range enhanced the formation of insoluble protein aggregates, whereas $(Ser)_8$, and $(Aia)_8$, did not form aggregates. Our results indicate that the formation of protein aggregates in GST-polyglutamine proteins is specifically mediated by the polyglutamine repeat sequence within their protein structure. Our study may contribute to the understanding of the molecular mechanism of the formation of protein aggregates in neurodegenerative disorders and the development of preventative strategies.

• Journal of the Korean Magnetic Resonance Society
• /
• v.15 no.2
• /
• pp.175-186
• /
• 2011

Amyloid fibrils have long been known to be the well known ${\alpha}$-helix to ${\beta}$-sheet transition characterizing the conversion of cellular to scrapie forms of the prion protein. A very short sequence of Yeast prion-like protein, GNNQQNY (SupN), is responsible for aggregation that induces diseases. KSI-fused tandem repeats of SupN vector are constructed and used to express SupN peptide in Escherichia coli (E.Coli). A method for a production, purification, and cleavage of tandem repeats of recombinant isotopically enriched SupN in E. coli is described. This method yields as much as 20 mg/L of isotope-enriched fusion proteins in minimal media. Synthetic SupN peptides and $^{13}C$ Gly labeled SupN peptides are studied by Congo Red staining, Birefringence and transmission electron microscopy to characterize amyloid fibril formation. To get a better understanding of aggregation-structure relationship of 7 residues of Yeast prion-like protein, the change of a conformational structure will be studied by $^{13}C$ solid-state nmr spectroscopy as powder of both amorphous and fibrillar forms.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.11
• /
• pp.1479-1483
• /
• 2004

Milk protein polymorphisms were genotyped by polyacrylamide gel electrophoresis (PAGE) from 109 Maiwa and 100 Jiulong yaks. The relationships between milk protein polymorphisms and 3 milking traits were studied. The results showed that $\beta$-CN, $\kappa$-CN and $\alpha$-La were monomorphic, and ${\alpha}_{s1}$-CN and $\beta$-Lg were polymorphic, with ${\alpha}_{s1}$-CN D and $\beta$-Lg E as dominant genes, respectively. The frequencies of ${\alpha}_{s1}$-CN D were 0.8073 and 0.6000 in two populations and $\beta$-Lg E were 0.9770 and 0.9700. The mean heterozygosities were 0.1021 and 0.1867 in the two populations. No significant effects on milking traits and milk protein compositions were observed except for ${\alpha}_{s1}$-CN locus on fat percentage in Jiulong yak.