Pathogenic Vibrio spp. Isolated from the Gwangan Beach of Busan, 2002

Park, Mi-Yeon,Kim, Hyun-Jin,Chang, Dong-Suck 한국수산학회 2003 Fisheries and Aquatic Sciences Vol.6 No.3

Fifty four strains of pathogenic vibrios were isolated from the Gwangan Beach from May to October, 2002. The isolated vibrios were composed of 7 different species: Vibrio parahaemolyticus, V. cholerae non-0 1, V. alginolyticus, V. vulnzjicus, V. hollisae, V. jluvialis, and V. mimicus. In the detection rate, V. parahaemolyticus was most predominant as 46% (25/54). From the isolated strains, only 25 strains have hemolytic activity or 25 strains only proteolytic activity on agar plates. Eleven strains showed both hemolytic and proteolytic activity. No strains showed urease activity. All strains of V. parahaemolyticus did not show hemolytic activity, while V. cholerae non-0 I strains showed P hemolytic activity. Kanagawa phenomena of pathogenic vibrios did not accord with hemolytic activity of the culture supernatant at the late log phase. Some strains showed high hemolytic activity despite having proteolytic activity, but some weak hemolytic activities despite having no proteolytic activity.

Structure-Antifungel Activity Relationships of Cecropin A Hybrid Peptides against Trichoderma sp.

Shin, Song-Yub,Lee, Dong-Gun,Lee, Sung-Gu,Kim, Kil-Lyong,Lee, Myung-Kyu,Hahm, Kyung-Soo The Microbiological Society of Korea 1997 The journal of microbiology Vol.35 No.1

The hybrid peptides, CA-ME, CA-MA and CA-BO, with the N-terminal sequence 1-8 of cecropin A and the N-terminal sequences 1-12 of melittin, magainin 2 and bombinin, respectively, have more improved antibacterial activities. CA-MA was found to have stronger antifungal activity against Trichoderma sp than other hybrid peptides and their parental peptides. In order to elucidate the relationships between the peptide structure and antifungal activity, several analogues of CA-MA or CA-BO were also designed and synthesized by the solid phase method. An tifungal activity was measured against T. reesei and T. viride, and hemolytic activity was measured by a solution method against human red blood cells. The residue 16 of CA-MA, Ser, was found to be important for antifungal activity. When the residue was substituted with Leu, showed powerful antifungal activity was dramatically decreased. CA-MA, P1, P4 and P5 designed in this study showed powerful antifungal activity against T. reesei and T. viride with low hemolytic activity against human red blood cells. These hybrid peptides will be potentially useful model to further design peptides with powerful antifungal activity for the effective therepy of fungal infection and understand the mechanisms of antifungal actions of hybrid peptides.

Bioactivity and molecular characterization of bombolitins from Bombus ardens, B. consobrinus, B. terrestris and B. ussurensis

윤경재,박영한,고영호,이시혁 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.4

Biological activities of bombolitins from Bombus ardens, B. consobrinus, B. terrestris and B. ussurensis (bombolitins A, C, T and U, respectively) were examined using hemolytic, anti-microbial, anti-fungal and anti-tumor activity assays. Among the four bombolitins tested, bombolitin T showed the highest hemolytic and anti-tumor activities. All bombolitins exhibited strong anti-microbial and anti-fungal activities, and bombolitin A specifically possessed the highest anti-microbial activity against the Gram-negative bacteria Escherichia coli. Circular dichroism spectrometry analysis revealed that all four bombolitins had over 61.7% and 45.5% of α-helicity in 30 mM sodium dodecyl sulfate and 50% trifluoroethanol buffers, respectively, which form lipid-membrane-mimicking environments. Bombolitin T showed the lowest IC50 values of 8.5 μM and 8.8 μM against SK-OV-3 and NIHOVCAR- 3 cell lines, respectively, after 72 h of treatment, but its relative hemolytic activity at a concentration of 200 μM was 2.3-fold higher than that of 0.1% Triton X-100. To design selective anti-tumor peptides with reduced hemolytic activity, bombolitin T was chosen as a model to be modified by amino acid substitution. The lowest hemolytic activity was observed with the addition of a hydrophobic amino acid (Ile14) to the C-terminal end as well as the substitution of Ser10 and Leu12 with a hydrophobic amino acid (Leu) and positively charged amino acid (Lys), respectively. This finding implies that the balanced substitution of hydrophobic and positively charged amino acids could affect the cytotoxicity. This study provides new information on the properties of antitumor peptides in the venoms of bumblebees and the basic approaches for peptide design tools for the reduction of cytotoxicity

Bioactivity and molecular characterization of bombolitins from <i>Bombus ardens</i>, <i>B. consobrinus</i>, <i>B. terrestris</i> and <i>B. ussurensis</i>

Yoon, Kyungjae Andrew,Park, Young Han,Koh, Young Ho,Lee, Si Hyeock 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol. No.

<P>Biological activities of bombolitins from Bombus ardens, B. consobrinus, B. terrestris and B. ussurensis (bombolitins A, C, T and U, respectively) were examined using hemolytic, anti-microbial, anti-fungal and anti-tumor activity assays. Among the four bombolitins tested, bombolitin T showed the highest hemolytic and anti-tumor activities. All bombolitins exhibited strong anti-microbial and anti-fungal activities, and bombolitin A specifically possessed the highest anti-microbial activity against the Gram-negative bacteria Escherichia coli. Circular dichroism spectrometry analysis revealed that all four bombolitins had over 61.7% and 45.5% of alpha-helicity in 30 mM sodium dodecyl sulfate and 50% trifluoroethanol buffers, respectively, which form lipid-membrane-mimicking environments. Bombolitin T showed the lowest IC50 values of 8.5 mu M and 8.8 mu M against SK-OV-3 and NIH-OVCAR-3 cell lines, respectively, after 72 h of treatment, but its relative hemolytic activity at a concentration of 200 mu M was 2.3-fold higher than that of 0.1% Triton X-100. To design selective anti-tumor peptides with reduced hemolytic activity, bombolitin T was chosen as a model to be modified by amino acid substitution. The lowest hemolytic activity was observed with the addition of a hydrophobic amino acid (Ile(14)) to the C-terminal end as well as the substitution of Ser(10) and Leu(12) with a hydrophobic amino acid (Leu) and positively charged amino acid (Lys), respectively. This finding implies that the balanced substitution of hydrophobic and positively charged amino acids could affect the cytotoxicity. This study provides new information on the properties of antitumor peptides in the venoms of bumblebees and the basic approaches for peptide design tools for the reduction of cytotoxicity.</P>

Characterization of biological activities of bombolitins from bumblebees Bombus ardens and Bombus ussurensis

Kyungjae Andrew Yoon,Kyungmun Kim,A-Young Kim,Young Han Park,Woo Young Bang,Chang Hwan Bae,Joo-Hong Yeo,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2016 한국응용곤충학회 학술대회논문집 Vol.2016 No.10

Bombolitin is a venom peptide originally isolated from bumblebees and possesses various biological activities, including hemolytic activity. Bombolitins exhibit amphipathic α-helical structure in lipid-membrane-mimicking environments. To investigate their pharmacological and toxicological properties, anti-tumor, anti-microbial and cytotoxic activities of bombolitins from Bombus ardens and Bombus ussurensis were evaluated. Bombolitins of the two species exhibited extremely high anti-tumor activity against ovarian tumor cells SK-OV-3 and NIH; OVCAR-3 at 25-50 μM, which is 2-fold more potent than other wasp venom peptides studied to date (Yoon et al., 2015; Yoon et al., 2016). The two bombolitins showed significantly high antimicrobial activity. However, bombolotin of B. ussurensis showed no antimicrobial activity against Escherichia coli. In addition to their high levels of anti-tumor activity, bombolitins showed considerable levels of hemolytic activity. Thus, to utilize bombolitins as a potential candidate for anti-tumor peptide drugs, further studies for reducing cytotoxic properties of bombolitns is essential.

Pathogenic Factors of Vibrio spp. Isolated from Seawater of Gwangan Beach in Busan

Park, Mi-Yeon,Kim, Hyun-Jin,Choi, Seung-Tae,Oh, Eun-Gyong,Chang, Dong-Suck 한국수산학회 2002 Fisheries and Aquatic Sciences Vol.5 No.3

The authors identified 68 Vibrio strains from Gwangan beach seawater from June to October in 2001. We identified them as 19 strains of Vibrio alginolyticus, 15 strains of V. fluvialis and just one strain of V. hollisae. They showed their typical biochemical characteristics by API 20E kit (bioMe´rieux), respectively. It was examined whether their cultural supernatants had enzymatic activities such as hemolysin, protease or urease. The 46 strains showed hemolytic activities and/or protease activities. But we could not find any strain which had urease activity. All isolates of V. cholerae non O1 showed β hemolysis. The other showed α hemolysis or did not show clear zones on sheep blood agar plates. These results of Kanagawa phenomenon were not always correspondant with hemolytic activities of cultural supernatants at late log phase. Some strains had higher hemolytic activities despite of showing protease activities on skim milk agar plates and in litmus milk media. On the other hand, some strains showed protease activities but did not show hemolytic activities. Therefore we could guess that there were the relationships between hemolysins and proteases produced by pathogenic vibrios.

CRAMP ANALOG HAVING POTENT ANTIBIOTIC ACTIVITY WITHOUT HEMOLYTIC ACTIVITY

Kang, Shin-Won,Lee, Dong Gun,Yang, Sung-Tae,Kim, Yangmee,Kim, Jae Il,Hahm, Kyung-Soo,Shin, Song Yub 부산대학교 유전공학연구소 2002 분자생물학 연구보 Vol.18 No.-

CRAMP-18 is an 18-residue functional region, corresponding to residues 16-33 of a mouse-derived antibiotic peptide CRAMP. To develop novel antibiotic peptides possessing strong antibiotic activity against bacterial, fungal and tumor cells without hemolytic activity, three analogs of CRAMP-18 were synthesized containing either Leu-or Lys-substitution. Leu-substitution ([L^1, 8]-CRAMP-18) in the hydrophobic helix face of CRAMP-18 induced a dramatic increase in antibiotic activity without a significant increase in hemolytic activity. Lys-substitution ([K^2, 13]-CRAMP-18 or [K9, 16]-CRAMP-18) in the hydrophilic helix face produced a smaller response. Therefore, [L^1, 8]-CRAMP-18 may be an attractive candidate for developing novel peptide antibiotics.

CRAMP Analogues Having Potent Antibiotic Activity against Bacterial, Fungal, and Tumor Cells without Hemolytic Activity

Shin, Song Yub,Kang, Shin-Won,Lee, Dong Gun,Eom, Soo Hyun,Song, Woo Keun,Kim, Jae Il 부산대학교 유전공학연구소 2000 분자생물학 연구보 Vol.16 No.-

CRAMP-18 (GEKLKKIGQKIKNFFQKL) is the anti-bacterial sequence derived from CRMAP, a member of cathelicidin-derived antimicrobial peptides. To develop the novel antibiotic peptides useful as therapeutic drugs requires strong antibiotic activity against bacterial and fungal cells without hemolytic effect. To this goal, the analogues were designed to increase only net positively charge by Lys-substitution of positions 2, 9, 13, or 16 at the hydrophilic helix face of CRAMP-18 without any change at the hydrophobic helix face. In particular, Lys-substitution (K^2-CRAMP-18) of position 2 in CRAMP-18 induced the enhanced antibiotic activity without any increase in hemolysis. Thus, this peptide may provide a useful template for the design novel antibiotic peptides for the treatment of infectious diseases. Additional CD spectra studies suggested that the α-helical structure of the peptides plays an important role in killing bacterial and fungal cells, but the increase of α-helical content is less connected with the enhanced antibiotic activity.

Activity Optimization of an Undecapeptide Ana-logue Derived from a Frog-Skin Antimicrobial Pep-tide

원형식,Su-Jin Kang,최완수,이봉진 한국분자세포생물학회 2011 Molecules and cells Vol.31 No.1

While natural antimicrobial peptides are potential therapeutic agents, their physicochemical properties and bioactivity generally need to be enhanced for clinical and commercial development. We have previously developed a cationic, amphipathic α-helical, 11-residue peptide (named herein GA-W2: FLGWLFKWASK-NH_2) with potent antimicrobial and hemolytic activity, which was derived from a 24-residue, natural antimicrobial peptide isolated from frog skin. Here, we attempted to optimize peptide bioactivity by a rational approach to sequence modification. Seven analogues were generated from GA-W2, and their activities were compared with that of a 12-residue peptide, omiganan, which is being developed for clinical and commercial applications. Most of the modifications reported here improved antimicrobial activity. Among them, the GA-K4AL (FAKWAFKWLKK-NH2) peptide displayed the most potent antimicrobial activity with negligible hemolytic activity, superior to that of omiganan. The therapeutic index of GAK4AL was improved more than 53- and more than 31-fold against Gram-negative and Gram-positive bacteria, respectively, compared to that of the starting peptide, GAW2. Given its relatively shorter length and simpler amino acid composition, our sequence-optimized GA-K4AL peptide may thus be a potentially useful antimicrobial peptide agent.

무우자갈버섯(Hebeloma crustulineforme)을 위시한 한국산 담자균류 46종의 용혈활성 검색

양희정,정수현,김진향,정경수 충남대학교 약학대학 의약품개발연구소 1997 藥學論文集 Vol.13 No.-

To elucidate the hemolytic toxicities of Korean basidiomycetes, the cold-water extracts of 46 wild mushrooms were tested for hemolytic activity on mouse erythrocytes. Twelve species including Amanita esculenta, A. griseofarinosa, A. longistriata, A. melleicepe, A. phalloides, A. rubescence, A. spissacea, A. vaginata, Hebeloma crustuliniforme, Lepiota cygnea, Macrolepiota procera, and Oudemansiella platyphylla showed strong hemolytic activities. Of these, A. griseofarinosa and H. crustuliniforme showed no decrease in their hemolytic activity even after five minutes' boiling.