• Asian-Australasian Journal of Animal Sciences
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• v.4 no.4
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• pp.369-375
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• 1991

A series of in vitro incubation studies with washed rumen bacteria were conducted to determine the influence of incubation time and concentrations of peptides, alanine, ammonia nitrogen and carbohydrate on the rate of peptide disappearance and on bacterial growth. Disappearance rate of tri-alanine (ala3) under various conditions was between 30.6 and $58.2mg\;hr^-$ per gram bacterial dry matter. Ala3 was removed from the incubation medium in an almost linear fashion as incubation time and ala3 concentration was increased. Washed rumen bacteria utilized ala3 faster than di-l-alanine (ala2) at all concentrations. Adding 9mM carbohydrate significantly increased ala3 disappearance, but level of ammonia nitrogen had no influence on ala3 disappearance. The presence of alanine in the medium significantly lowered ala3 utilization by rumen bacteria. Bacterial dry matter and nitrogen growth yield were not influenced by alanine and peptides when incubation medium already contained a sufficient level of ammonia nitrogen. Increased ammonia nitrogen in the presence of ala3 did not stimulate bacterial growth. Carbohydrate significantly increased bacterial dry matter and nitrogen growth as expected. Results indicate that the rate of peptide utilization by rumen bacteria may be altered by type and concentration of peptides, and energy supply, and this may be mediated through changes in numbers and type of bacteria.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.2
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• pp.282-288
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• 2009

The present paper describes the effects of pressure and post-mortem aging treatments on in situ proteasome activity in rabbit and bovine skeletal muscles. Synthetic peptide hydrolyzing activity of rabbit proteasomes remained in the muscle after exposure to pressures up to 100 MPa. However, when a pressure of 400 MPa or more was applied, proteasomes were markedly inactivated. The extraction of proteasomes from excessively pressurized muscle appeared to be difficult. Proteasomes in aged muscle remained relatively stable throughout the aging process, with activity after 168 h (7 days) being 35%, 48%, 53% and 31% of the 0 h post-mortem LLVY, LSTR, AAF and LLE total hydrolyzing activities, respectively. The synthetic peptide hydrolyzing activities of bovine muscle proteasomes were similar to those of rabbit skeletal muscle proteasomes. The results suggest that synthetic peptide hydrolyzing activity remains in muscle exposed to relatively low pressures. Furthermore, it is known that high-pressure treatment induces fragmentation of myofibrils, modification of actin-myosin interaction and activation of intramuscular proteinases, cathepsins and calpains. Thus, proteasomes are probably involved in the tenderization process in combination with other intramuscular proteinases under high-pressure conditions. Our findings confirmed that proteasomes play a role in meat tenderization induced by high-pressure treatment or aging.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.753-759
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• 2002

$\beta$-Cyclodextrin glucanotransferase ($\beta$-CGTase) was overproduced extracellularly using recombinant E. coli by transforming the plasmid pECGT harboring a secretive signal peptide. The $\beta$-CGTase gene of alkalophilic Bacillus firmus var alkalophilus was inserted into the high expression vector pET20b(+) containing a secretive pelB signal peptide, and then transformed into E. coli BL2l(DE3)pLysS. The optimum culture conditions fer the overproduction of $\beta$-CGTase were determined to be TB medium containing 0.5% (w/v) soluble starch at post-induction temperature of $25^{\circ}C$. A significant amount of $\beta$-CGTase, up to 5.83 U/ml, which was nine times higher than that in the parent strain B. firmus var. alkalophilus, was overproduced in the extracellular compartment. A pH-stat fed-batch cultivation of the recombinant E. coli was also performed to achieve the secretive overproduction of $\beta$-CGTase at a high cell density, resulting in production of up to 21.6 U/ml of $\beta$-CGTase.

• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1429-1433
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• 2006

Application of recombinant protein production and particularly their isotopic enrichment has stimulated development of a range of novel multidimensional heteronuclear NMR techniques. Peptides in most cases are amenable to assignment and structure determination without the need for isotopic labeling. However, there are many cases where the availability of $^{15}N$ and/or $^{13}C$ labeled peptides is useful to study the structure of peptides with more than 30 residues and the interaction between peptides and membrane. CRAMP (Cathelicidin-Related AntiMicrobial Peptide) was identified from a cDNA clone derived from mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptides. CRAMP was successfully expressed as a GST-fused form in E. coli and purified using affinity chromatography and reverse-phase chromatography. The yield of the CRAMP was 1.5 mg/l 1. According to CD spectra, CRAMP adopted ${\alpha}$-helical conformation in membrane-mimetic environments. Isotope labeling of CRAMP is expected to make it possible to study the structure and dynamic properties of CRAMP in various membrane systems.

• Bulletin of the Korean Chemical Society
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• v.30 no.8
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• pp.1839-1844
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• 2009

A 20-residue hybrid peptide CA(1-8)-MA(1-12) (CAMA) incorporating residues 1-8 of cecropin A (CA) and residues 1-12 of magainin 2 (MA) has high antimicrobial activity without toxicity. To investigate the effects of the total positive charges of CAMA on the antibacterial activity and toxicity, a hybrid peptide analogue (CAMA-syn) was designed with substitutions of $Ile^{10}\;and\;Ser^{16}$ with Lys. According to CD spectra, structure of CAMA-syn with increase of cationicity was very similar to that of CAMA in DPC micelle. CAMA-syn showed antimicrobial activity similar with CAMA while CAMA-syn has no hemolytic activity and much lower cytotoxicity against RAW 264.7 macrophage cells than CAMA. Also, CAMA and CAMA-syn significantly inhibited NO production by LPSstimulated RAW264.7 macrophage at 10.0∼20.0 $\mu$M. CAMA-syn displayed salt resistance on antimicrobial activity against Escherichia coli at the physiological concentrations of $CaCl_2\;and\;MgCl_2$. The combination studies of peptides and antibiotics showed that CAMA-syn has synergistic effects with synthetic compound and flavonoid against Enterococcus faecalis and VREF. CAMA-syn can be a good candidate for the development of new antibiotics with potent antibacterial and synergistic activity but without cytotoxicity.

• KSBB Journal
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• v.8 no.3
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• pp.209-216
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• 1993

For the production of biodegradable polymers from microorganisms a bacterial strain producing a biopolymer was isolated from soil. The bacteriological characteristics of this strain and physicochemical Properties of the biopolymer produced were investigated. The bacterial strain was identified as an alkalophilic Alcaligenes sp. The Purified biopolymer treated with cetylpridinium chloride and acetone was identified as an acidic biopolymer having carboxyl groups and showed strong UV absorbance (at 210nm). The biopolymer was composed of 100% glutamic acid and glutamic acid existed as $\gamma$-polyglutamic acid($\gamma$-PGA) in the form of the $\gamma$-peptide bond. The equivalent weight of this $\gamma$-PGA was estimated about 350, indicating that one acidic fraction per 2.7 residue of $\gamma$-polyglutamic acid existed. The molecular weight was $6.5{\times}10^5$ Daltons.

• Journal of Microbiology and Biotechnology
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• v.34 no.8
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• pp.1680-1687
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• 2024

The diverse pharmacological properties of edible bird's nest (EBN) have been elucidated in recent years; however, investigations into its antibacterial effects are still limited. In the present study, we explored the antibacterial activity of a peptide-rich extract of EBN against Staphylococcus aureus, a notorious pathogen. The EBN extract (EEE) was prepared by soaking EBN in 80% ethanol for 2 days at 60℃. Biochemical analyses showed that peptides at the molecular weight range of 1.7-10 kDa were the major biochemical compounds in the EEE. The extract exhibited strong inhibition against S. aureus at a minimum inhibitory concentration (MIC) of 125 ㎍/ml and a minimum bactericidal concentration (MBC) of 250 ㎍/ml. This activity could be attributed to the impact of the extract on cell membrane integrity and potential, biofilm formation, and reactive oxidative species (ROS) production. Notably, the expression of biofilm- and ROS-associated genes, including intercellular adhesion A (icaA), icaB, icaC, icaD, and superoxide dismutase A (sodA), were deregulated in S. aureus upon the extract treatment. Our findings indicate a noteworthy pharmacological activity of EBN that could have potential application in the control of S. aureus.

• Journal of Applied Biological Chemistry
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• v.60 no.2
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• pp.149-153
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• 2017

Contamination and growth of Trichoderma, a green mold, on the oak log and wooden chip or sawdust media can severely inhibit the growth of oak mushroom. Chemicals including pesticides and antibiotics are generally not allowed for the control of green mold disease during mushroom cultivation. In this study, bacterial pathogens causing blotch disease on the oyster mushrooms were isolated and their peptide toxins were purified for the control of green mold disease. Strains of Pseudomonas tolaasii secret various peptide toxins, tolaasin and its structural analogues, having antifungal activities. These peptides have shown no effects on the growth of oak mushrooms. When the peptide toxins were applied to the green mold, Trichoderma harzianum H1, they inhibited the growth of green molds. Among the 20 strains of peptide-forming P. tolaasii, strong, moderate, and weak antifungal activities were measured from 8, 5, and 7 strains, respectively. During oak mushroom cultivation, bacterial culture supernatants containing the peptide toxins were sprayed on the aerial mycelia of green molds grown on the surface of sawdust media. The culture supernatants were able to suppress the fungal growth of green molds while no effect was observed on the mushroom growth and production. They changed the color of molds from white aerial mycelium into yellowish dried scab, representing the powerful anti-fungal and sterilization activities of peptide toxins.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.92-97
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• 1996

Portage kinetic constants of peptide transport can be measured by competitive spectrophotometry. The kinetic constants of L-Glu-L-Glu transport in Escherichia coli were ascertained using L-Phe-L-3-thia-Phe (PSP) as a detector. Since the production of thiophenol upon intracellular hydrolysis of PSP was competitively inhibited by L-Glu-L-Glu, it was able to compute the kinetic constants of L-Glu-L-Glu using this method. The resulted data were in agreement with the values obtained by the method of Michaelis-Menten kinetics. The potential of this method was examined against dipeptide transport systems in various microorganisms. These results strongly suggest that the overall properties of individual systems for dipeptide transports can be easily characterized by competitive spectrophotometry.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.669-671
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• 2000

Antimicrobial cationic peptides have attracted increasing research and clinical interest as a natural antibiotics due to their broad spectrum of antimicrobial activites and the rapid development of multidrug-resistant pathogenic microorganisms. In this study, first, we synthesized artificial fusion partner and cationic peptide genes (lactoferricin, magainin, protegrin-1, and indolicidin). Second, we constructed recombinant expression vectors and then transformed Pichia pastoris. Finally, expressed cationic peptides were purified and tested for their antimicrobial activites. Antimicrobial activity has been tested upon the appearance of clearing zone on the plate with the lawn of gram negative E.coli XL- I blue and garm positive Staphylococcus aureus. Protegrin-1 and Indolicidin have apparant activity of cationic peotides. This fusion technique may lead to a general and suitable tool for production of pure antimicrobial cationic peptides in Pichia pastoris.