Purification and Characteristic Properties of Ribonuclease H from Sea Urchin, Hemicentrotus pulcherrimus

류병호,이병호,박형숙,하미숙,김동석,Ryu, Beung-Ho,Lee, Beung-Ho,Park, Hyeong-Suck,Ha, Mi-Suck,Kim, Dong-Seuk 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.3

말똥 성게를 인공적으로 수정시킨 후, 칼럼 크로마토그래피법으로 분리 정제하여 리보뉴크레아제 $H_1$과 리보뉴크레아제 $H_2$를 얻었다. 리보뉴크레아제 $H_1$의 분자량은 약 68,000이었고, 리보뉴크레아제 $H_2$의 분자량은 약 44,000 이었다. 리보뉴크레아제 $H_1$는 약알카리성에서, 리보뉴크레아제 $H_2$는 알카리성에서 활성이 높았다. 이들 효소는 모두 2가이온인 마그네슘이온 ($Mg^{2+}$), 망간이온($Mn^{2+}$)과 그리고 염의 일정농도에서 활성을 나타내고 있으며, N-에칠 말례이마이드에 감수성이 높았다. 그리고, 리보뉴크레아제 $H_1$과 $H_2$는 단일가닥과 이중가닥의 활성은 없었으나, RNA-DNA hydrid는 급히 저하 시켰다. From the sea-urchin, Hemicentrotus pulcherrimus, we have purified by two column chromatographic steps two ribonuclease H (RNase H) activities of RNase $H_1$ and RNase $H_2$ in homogeneous form. RNase $H_1$ was molecular weight around 68,000 and RNase $H_2$ was around 44,000 by Sephadex G-75 gel filteration and SDS-polyacrylamid gel electrophoresis. RNase $H_1$ was optimal active as well as slightly alkaline pH and RNase $H_2$ was as alkaline pH. These enzymes shown to be a function of the divalent metal ion $Mg^{2+}$, $Mn^{2+}$ and salt employed as activator, also were sensitive -SH reagent N-ethylmaleimide. RNase $H_1$ and RNase $H_2$ were shown no activity with single or double stranded RNA but rapidly hydrolysis the RNA-DNA hybrid.

카드뮴 축적 변이주인 Pseudomonas maltophilia H - 8M의 특성

류병호(Beung-Ho Ryu),노명훈(Myung-Hoon Rho),정수자(Su-Ja Jung),배기철(Ki-ChuI Bae) 한국식품영양과학회 1992 한국식품영양과학회지 Vol.21 No.1

본 연구는 cadmium의 축적균주인 Pseudomonas maltophilia H-8을 MNNG로 처리하여 cadmium축적능이 가장 우수한 변이균주인 Pseudomonas maltophilia H-8M을 얻었고 그 특성을 조사하였다. 변이주 Pseudomonas maltophilia H-8M은 카드뮴농도 1,000ppm에서 36시간 배양했을 시 대조구에 비해 약 80%정도의 성장율을 나타내었다. 카드뮴 이외의 중금속의 첨가에 따른 생육상은 Pb^(2+), Zn^(2+), Hg^(2+), Cu^(2+), Cr^(2+), Co^(2+)등에는 별다른 성장 억제는 없었으나 Sn^(2+)에서는 생육억제가나타났다. 또한 50ppm의 카드뮴을 배지에 첨가 하였을 때 균체내 카드뮴 축적율은 52.92%로 가장 높게 나타났으며 축적부위는 세포벽에 80.3%의 축적율을 나타내었다. This study was carried out to investigate the characteristics of a mutant, Pseudomonas maltophilia Ho8M selected with the treatment of Pseudomonas maltophilia H-8 by N-methyl-N-nitro-N-nitrosoguanidine (MNNG). This mutant showed highest ability of cadmium accumulation. The growth rate of Pseudomonas maltophilia H-8M showed about 80% in 1000ppm Cd containing medium when compare with control for 36h at 30℃. Pseudomonas maltophilia H-8M not inhibited on the growth in addition of various heavy metal such as Hg^(2+), Zn^(2+), Pb^(2+), Cu^(2+), Cr^(2+) and Co^(2+), but inhibited in Sn^(2+) containing medium, respectively. Pseudomonas maltophilia H-8M was accumulated the highest cadmium level of 62.3% on whole cell in the medium containing 50 ppm and 80% of accumulated cadmium was distributed in the cell wall.

Antioxidative activity of flavonoid rich extract of Oenothers odorate Jacquin on oxidation of low density lipoprotein

류병호,김희숙,조경자,Ryu, Beung-Ho,Kim, Hee-Sook,Cho, Kyung-Ja Korean Society of Life Science 2002 생명과학회지 Vol.12 No.3

There is growing interest ill understanding the role and mechanisms of flavonoid as antioxidant on LDL. The antioxidative activity of flavonoid typically present in Oenothers odorate Jacquin was investigated in vitro using a human LDL oxidation assay. In present work, LDL was incubated with increasing concentrations of extracts of extracts of Oenothers odorate Jacquin and LDL oxidation was started by adding CuSO$_4$to the media. Substances in leaves extracts of Oenothers odorate Jacquin are capable of inhibiting the initiation and the propagation of LDL oxidation. They inhibit LDL oxidation, monitored by thiobarbituric acid-reactive substances(TBARS), as well as modification as shown through direct measurement of electrophoretic mobility, diene conjugates. Inhibition is a dose dependent effect that becomes already apparent at concentration of extracts as low as 40$\mu\textrm{g}$/mL. Inhibition is almost complete at 80$\mu\textrm{g}$/mL. Extracts of Oenothers odorate Jacquin were more potent antioxidative activity than either ascorbic acid and dl-a -tocopherol. 달맞이꽃 추출물로써 항산화활성을 알아보기 위하여 사람 low densty lipoprotein(LDL)에 대하여 실험하였다. 사람 LDL에 CuSO$_4$를 첨가한 후 배양하면서 달맞이꽃 추출을 일정 농도씩 증가시키면서 첨가하여 배양하였다. 달맞이꽃 추출물로서 LDL의 산화에 대한 개시와 연쇄 반응에서 thiobarbituric acid reactivy substances (TBARS)와 전기 영동 및 공액 2중 결합으로 측정하였다. 달맞이꽃 추출물을 LDL에 첨가한 후 macrophage J774로써 배양한 결과 TBARS값은 용량 의존형의 항산화 활성을 나타내었으며 전기 영동 이동상에서도 대조구에 비하여 각 시험구에서 LDL의 산화를 억제하였다. 또 공액 2중 결합에 대한 실험에서 용량 의존형산화 억제 효과를 나타내었다. 달맞이꽃 추출물의 항산화 활성은 40 $\mu\textrm{g}$/ml의 농도에서도 항산화 효과가 있었으나 80$\mu\textrm{g}$/mL의 농도에서는 거의 완전한 억제효과가 있었다. 그리고 비타민 C 및 $\alpha$-dl-tocopherol과 비교한 결과 달맞이들 추출물이 약간 높은 항산화 활성을 나타내었다.

엉겅퀴에서 분리정제한 silybin의 Human Low Density Lipoprotein 수식에 대한 항산화 효과

류병호,김희숙,이백천,이홍수,Ryu, Beung-Ho,Kim, Hee-Suck,Lee, Baek-Cheon,Lee, Hong-Su 한국생명과학회 1998 생명과학회지 Vol.8 No.4

This study was initiated to antioxidant activity of silybin on oxidation of human low density lipoproteins(LDL). Siltbin was extracted from Silybum marianum by the combination of fractionation and it was futher purified by silica gel column chromatography, and isolated active substances were identified silybin by IR, NMR and GC-MS. siltbin inhibited the ozidation of human low density lipoprotein(LDL) mediated by 5$^{\mu}$m CU $^{2+}$ ion in a dose dependent manner. LDL oxidation by congugated dines formation was completely inhibited by silybin at a concentration of 5$^{\mu}$M. The results provide a possibility that silybin might protect LDL against oxidation in atherosclerotic lessions.

Aspergillus oryzae A-5로부터 Low Density Lipoprotein(LDL)의 산화에 대한 항산화 효과

류병호,김동석,조경자,이홍수,진성현,Ryu, Beung-Ho,Kim, Dong-Suck,Cho, Kyung-Ja,Lee, Hong-Su,Jin, Sung-Hyun 한국생명과학회 1997 생명과학회지 Vol.7 No.4

Antioxidative activity of fraction extracted from cultivation of Aspergillus sp. A-5 against oxidation of human low density lipoprotein(LDL) was investigated. Fractions of Aspergillus sp. A-5 cultivation was sucessively purified with ethyl acetate and silica gel column chromatography. The concentration of fraction 4 inhibited Cu$^{2+}$-induced oxidation of LDL almost completely. Band 3 isolated by the further purification of fraction 4 was higher than that of same concentration of $\alpha$-tocopherol, BHA and BHT. The elcetrophoretic mobility of oxidized LDL by addition of Band 3 was faster than that of native LDL, but slower than that of oxidzed LDL. It is concluded that fraction of Aspergillus cultivation contained antioxidants with the capacity to inhibit oxidative modification of LDL.

카페인 분해균주의 분리 및 특성

류병호,주신회,Ryu, Beung-Ho,Ju, Sin-Hae 한국식품과학회 1992 한국식품과학회지 Vol.24 No.3

Caffeine을 기질로 이용할 수 있는 균을 토양과 폐수에서 7종을 분리하고 그중 카페인 분해능이 가장 좋은 KS-5를 동정하여 Pseudomonas putida KS-5라고 명명하였다. P. putida KS-5의 생장 최적조건은 온도 $30^{\circ}C$, pH 7.0이었고 카페인 농도는 1.0%였다. P. putida KS-5는 plasmid DNA가 존재하였으며, 카페인 분해유전자가 plasmid에 존재할 수 있음을 알 수 있었다. 한편 이들 분해유전자의 유전적 특징을 규명하기 위한 curing test와 transformation에 필요한 marker를 찾아본 결과 각종 항생물질에 대하여 내성이 있었다. Several bacterial strains capable of degrading caffeine were isolated and studied for their biodegradation ability of the caffeine and some biochemical characteristics. The isolate KS-5 was identified as Pseudomonas putida and was designated as the P. putida KS-5. The optimum conditions were at $30^{\circ}C$, pH 7.0 and 1.0% caffeine. Agarose gel electrophoresis and curing experiment were found that the gene for caffeine degradation was encoded on the plasmid in P. putida KS-5 and that this strain was resistant to several antibiotics.

Desmutagenic Effect of Legumes and Plants Crude Saponins in Salmonella typhimurium TA 98

Beung-Ho Ryu(류병호),Beung-Ho Lee(이병호),Mi-Suck Ha(하미숙),Dong-Seuk Kim(김동석),Dong-Bun Sin(신동분),Ki-Du Nam(남기두) 한국식품과학회 1986 한국식품과학회지 Vol.18 No.5

해양에서 분리한 Streptomyces sp. BH-405 배양액의 항산화 효과

류병호(Beung-Ho Ryu),이영숙(Young-Sook Lee),양승택(Seong-Taek Yang) 한국생물공학회 2000 KSBB Journal Vol.15 No.2

Production of Mirin by Mutants of Aspergillus sp.

류병호(Beung-Ho Ryu),신동분(Dong-Bun Shin),빈재훈(Jae-Hoon Bin),박형선(Hyung-Sun Park) 한국식품영양과학회 1993 한국식품영양과학회지 Vol.22 No.5

본 연구는 품질이 우수한 미린을 생산하기 위하여 미린의 생산에 필요한 koji제조 균주를 검색하여 이들 중 ACPase활성이 높은 균주는 Aspergillus oryzae 9와 Aspergillus shirousamii IFO 6082를 얻었다. 이 두 균주는 UV조사에 의하여 변이주인 Aspergillus oryzae 9-12와 Aspergillus shirousamii IFO 6082-60을 얻었는데 acid carboxypeptidase의 활성은 친주에 비하여 2.6배 높았다. 이 균주로 재래식과 개량식 방법으로 미린을 제조한 결과 총당, 환원당 및 총질소의 양은 비슷하였으나 아미노태질소는 재래식보다 개량식이 높았고 수율도 높았다. 미린의 품질에 지대한 영향을 미치는 혼탁의 형성은 개량식에서 변이주로 만든 미린에서는 나타나지 않았다. To improve the quality of mirin, various molds were screened for mutants with high acid carboxypeptidase (ACPase) by the method of ultraviolet radiation. Mutants, Aspergillus oryzae 9-12 and Aspergillus shiroussamii 6082-60 showed activities of ACPase about 2-6 times higher than their parent strains. Aspergillus oryzae 9-12 and Aspergillus shirosamii 6082-60 were the most suitable strains for preparing koji in mirin by the conventional or improved methods. The results showed that total sugar, reducing sugar and total nitrogen were almost the same values in mirin prepared by both methods. The yield of mirin was higher in the improved method than in the conventional method. The clouding formation of mirin appeared in the conventional method; however, mirin prepared with the mutant koji by the improved method did not show clouding formation.

Cunninghamella blakesleeana IFO 4443의 변이주로부터 키토산의 생산

류병호(Beung-Ho Ryu),김희숙(Hee-Suck Kim),원용돈(Yong-Don Won),임복규(Bok-Gu Lim) 한국식품영양과학회 1997 한국식품영양과학회지 Vol.26 No.1

키토산을 생산하기 위하여 Cunninghamella blakesleeana IFO 4443을 자외선조사하여 생성능이 우수한 Cunninghamella blakesleeana IFO 4443-10의 변이주를 분리하였다. 본 변이주인 C. blakesleeana IFO 4443-10을 30℃에서 pH 4.5에서 3일간 발효하여 mycelia 발효액을 알칼리로 처리하여 chitosan을 얻었다. 이때 chitosan의 수율은 1012㎎/l로 높았으며 탈아세틸화는 84.6%이였다. A method for lab scale production and isolation of chitosan from mycelia of Cunninghamella blakesleeana IFO 4443 mutant was developed. Mutant of Cunninghamella blakesleeana IFO 4443-10 obtained by UV radiation was cultivated in 5L jar fermentor at 30℃ for 3days. The fungus were grown well at pH 4.5. Chitosan was readily extracted from mycelia walls with alkali treatment. The maximum yield of chitosan obtained was 1012㎎/L and degree of deacetylation was 84.6%.