혐기성 미생물에 의한 toluene과 xylene 분해 특성

조경숙,조원실,류희욱 한국냄새환경학회 2004 실내환경 및 냄새 학회지 Vol.3 No.2

Six kinds of the microbial consortia were obtained from the enrichment culture using the mixture gases of toluene and xylene as sole carbon sources and sulfate as a final electron acceptor. The degradation rates of the mixture gases by the consortia were determined, and the microbial structure in the consortia were characterized using 16S rDNA PCR and denaturing gradient gel electrophoresis (DGGE). The enriched consortia could simultaneously degrade toluene and xylene in the mixture gases. The toluene and xylene degradation rates in the mixture gases were 24.6∼49.1 and 2.8∼6.2 μmol L-1 d-1, respectively. Nine clones among 15 clones, isolated from the enriched consortia, were affiliated with the bacteria related to the degradation of hydrocarbons, chlorinated solvent, trichlorobenzene and TCE. These clones are expected to play important roles on the anaerobic degradation of toluene and xylene in the consortium. Toluene과 xylene의 혼합가스를 유일 탄소원으로 하고, sulfate를 최종전자수용체로 공급한 조건에서 농화 배양하여 얻은 6 종류의 미생물 농화배양액의 toluene과 xylene분해 특성을 조사하고 미생물 군집 특성을 분자생물학적 방법을 이용하여 분석하였다. 본 연구에서 얻은 농화배양액은 toluene과 xylene을 동시에 분해할 수 있었고, toluene과 xylene 분해속도는 각각 24.6∼49.1 및 2.8∼6.2 μmol L-1 d-1이었다. 농화배양액으로부터 분리한 15개의 clone 중에서 9개의 clone이 탄화수소화합물, 염소계 용매, trichlorobenzene 및 TCE 분해와 관련된 세균들과 유사성이 높은 것으로 동정되었는데, 이들 clone이 toluene 및 xylene 혼합가스의 혐기성 생분해에 관여하고 있을 것으로 사료되었다.

흰쥐 성장기간에 따른 Xylene의 독성에 관한 연구

이혜자(Hye-Ja Lee),이상희(Sang-Hee Lee),전태원(Tae-Won Jeon),이상일(Sang-Il Lee),윤종국(Chong-Guk Yoon) 대한의생명과학회 2000 Biomedical Science Letters Vol.6 No.3

실험동물에 있어서 연령 차이에 따라서 xylene 독성이 어떠한 차이가 있는지를 검토하는 일환으로 5주령 및 12주령 흰쥐에 50% m-xylene을 체중 100 g 당 0.25 ㎖씩 l회 투여한 다음 24시간 후에 처치하여 다음과 같은 결과를 얻었다. Xylene 투여로 인한 요 중 methylhippuric acid 함량은 5주령군이 12주령군에 비해 현저하게 높게 나타났다. 그리고 간조직 중 cytochrome P-450 함량은 대조군에 있어서 5주령군이 12주령군 보다 약 50% 정도 낮게 나타났으나 xylene 투여로 인한 cytochrome P-450 함량 증가율은 12주령군 보다 5주령군에서 높게 나타났다. 간 alcohol dehydrogenase 활성치도 대조군에 있어서는 5주령군이 12주령 보다 약 35% 정도 낮게 나타났으나 xylene 투여로 인한 본 효소의 활성 증가율은 5주령군에서 오히려 높게 나타났다. 그러나 간 aldehyde dehydrogenase 활성치는 대조군 및 xylene 투여군 모두 5주령과 12주령간에 유의한 차이를 나타내지 않았다. 한편 xylene 투여시 체중 당 간무게, 간조직 malondialdehyde 함량 및 혈청 ALT 활성 변동을 통하여 간손상 정도를 상호 비교 관찰하였을 때, 12주령군이 5주령 실험동물 보다 간손상이 다소 심하게 나타남을 알 수가 있었다. 이상 실험결과는 연령에 따라 xylene에 의한 간손상의 차이는 이물질의 생체내 대사율이 달리 나타나기 때문일 것으로 생각된다. To evaluate an effect of aging on the xylene toxicity, 50% m-xylene in olive oil (0.25 ㎖/100 g body wt.) was administered to 5 week and 12 week-old rats one times intraperitoneally and sacrificed at 24 hrs afterwards. The increasing rate of urinary m-methylhippuric acid concentration was higher in 12 week-old rats than 5 week-old rats by the treatment of m-xylene. On the liver function findings, i.e., liver weight/body weight (%), serum levels of ALT activity and hepatic malondialdehyde content, 12 week-old rats showed more severe liver injury than 5 weeks those in xylene-treated rats. And the hepatic cytochrome P-450 contents was higher in 12 weeks rats than those of in 5 week-old rats, but the increasing rate of that was lower in 12 week-old rats. Hepatic alcohol dehydrogenase activity was also higher in 12 week-old rats than in 5 week-old rats whereas the increasing rate of that was higher in 5 week-old than those in 12 week-old rats by the xylene treatment. Furthermore, the hepatic aldehyde dehydrogenase activities were no differences between the 5 and 12 week-old rats both in the control and xylene-treated group. In conclusion, age may influences upon the hepatotoxicity with xylene and it may be responsible for xylene metabolism in rats.

흰쥐에 Xylene 반복 투여가 Xylen의 대사에 미치는 영향

이혜자(Hye-Ja Lee),조현국(Hyun-Gug Cho),이상일(Sang-Il Lee),전태원(Tae-Won Jeon),윤종국(Chong-Guk Yoon) 대한의생명과학회 1999 Biomedical Science Letters Vol.5 No.1

실험동물에 xylene의 반복 투여가 이물질의 대사에 어떠한 영향을 미치는지를 알아보기 위하여 흰쥐에 m-xylene과 olive oil의 동량혼합액을 체중 100g당 0.25 ㎖씩 2일 간격으로 1, 4, 8, 12 및 16회 복강으로 투여한 다음 마지막 투여 24시간 후에 처치하여 다음과 같은 결과를 얻었다. 요 중 m-methylhippuric acid 함량은 m-xylene 4회 투여군의 경우 1회 투여군에 비하여 약 56%의 유의한 증가를 보였으며 이후 12회 투여까지 유사한 결과를 나타내었으나 m-xylene 16회 투여시에는 xylene 1회 투여군 치와 유사한 치로 감소되었다. 그리고 간조직의 microsomal aniline hydroxylase와 alcohol dehydrogenase 활성은 m-xylene 12회 투여시 까지는 대체적으로 점진적인 증가를 보였으나 이후 16회 투여시에는 12회 투여군에 비하여 유의한 감소를 보였다. 또한 aldehyde dehydrogenase 활성은 m-xylene 투여 회수에 비례해서 전 실험기간 동안 감소되었으며 특히 16회 투여군에서 본 효소활성의 현저한 감소를 보였다. 한편 본 실험조건에서 투여기간에 따른 전자현미경적 미세구조의 변화는 초기에 활면소포체의 증식이 보이다가 16회 투여군에서는 활면소포체가 감소되고 조면소포체가 증가되었다. 이상 실험결과는 흰쥐에 있어서 xylene 투여 회수에 따라서 요 중 m-methylhippuric acid의 농도 변동이 초래되며 이는 효소단백 유도에 따른 xylene 대사효소 활성 변동에 기인된 결과로 생각된다. To evaluate the effect of repeated treatment of xylene on its metabolism, m-xylene (0.25 ㎖ of 50% in olive oil/100 g body weight) has been intraperitoneally given to the rats 1, 4, 8, 12 and 16 times every other day. m-Xylene was once more administered to the animals after 24 hrs since last injection of it. And then the animals were sacrificed after 24 hrs. Four times xylene treated rats showed the significantly elevated urinary m-methylhippuric acid, compared to those treated with the single dose of m-xylene with the continued similiar high levels of urinary m-methylhippuric acid up to the animals pretreated 12 times and then those treated 16 times defined the significantly decreased urinary m-methylhippuric acid compared to those treated 12 times. On the other hand, hepatic aniline hydroxylase and alcohol dehydrogenase activities demonstrated a gradual increase from the first group to the 12 times xylene-treated animals, but those treated 16 times showed the significantly decreased value compared with the 12 times treated-group. And aldehyde dehydrogenase activities in rats treated with m-xylene 8, 12 or 16 times were significantly decreased compared to those pretreated one or four times. In the early stage of xylene administration, proliferation of SERs were seen whereas SERs were decreased and RERs were clearly increased in xylene-treated rats 16 times. These results indicate that the frequency of xylene injection may influence upon the changes in xylene metabolite, m-methylhippuric acid and it may be due to induction of xylene metabolizing enzymes.

흰쥐에 Xylene 반복 투여가 Xylene의 대사에 미치는 영향

전태원,이혜자,윤종국,이상일,조현국 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1999 Journal of biomedical laboratory sciences Vol.5 No.1

실험동물에 xylene의 반복 투여가 이물질의 대사에 어떠한 영향을 미치는지를 알아보기 위하여 흰쥐에 m-xylene과 olive oil의 동량혼합액을 체중 100g당 0.25ml씩 2일 간격으로 1, 4, 8, 12 및 16회 복강으로 투여한 다음 마지막 투여 24시간 후에 처치하여 다음과 같은 결과를 얻었다. 요 중 m-methylhippuric acid 함량은 m-xylene 4회 투여군의 경우 1회 투여군에 비하여 약 56%의 유의한 증가를 보였으며 이후 12회 투여까지 유사한 결과를 나타내었으나 m-xylene 16회 투여시에는 xylene 1회 투여군 치와 유사한 치로 감소되었다. 그리고 간조직의 microsomal aniline hydroxylase와 alcohol dehydrogenase 활성은 m-xylene 12회 투여시 까지는 대체적으로 점진적인 증가를 보였으나 이후 16회 투여시에는 12회 투여군에 비하여 유의한 감소를 보였다. 또한 aldehyde dehydrogenase 활성은 m-xylene 투여 회수에 비례해서 전 실험기간 동안 감소되었으며 특히 16회 투여군에서 본 효소활성의 현저한 감소를 보였다. 한편 본 실험조건에서 투여기간에 따른 진자현미경적 미세구조의 변화는 초기에 활면소포체의 증식이 보이다가 16회 투여군에서는 활면소포체가 감소되고 조면소포체가 증가되었다. 이상 실험결과는 흰쥐에 있어서 xylene 투여 회수에 따라서 요 중 m-methylhippuric acid의 농도 변동이 초래되며 이는 효소단백 유도에 따른 xylene 대사효소 활성 변동에 기인된 결과로 생각된다. To evaluate the effect of repeated treatment of xylene on its metabolism, m-xylene (0.25ml of 50% in olive oil/100 g body weight) has been intraperitoneally given to the rats 1, 4, 8, 12 and 16 times every other day. m-Xylene was once more administered to the animals after 24 hrs since last injection of it. And then the animals were sacrificed after 24 hrs. Four times xylene treated rats showed the significantly elevated urinary m-methylhippuric acid, compared to those treated with the singe dose of m-xylene with the continued similiar high levels of urinary m-methylhippuric acid up to the animals pretreated 12 times and then those treated 16 times defined the significantly decreased urinary m-methylhippuric acid compared to those treated 12 times. On the other hand, hepatic aniline hydroxylase and alcohol dehydrogenase activities demonstrated a gradual increase from the first group to the 12 times xylene-treated animals, but those treated 16 times showed the significantly decreased value compared with the 12 times treated-group. And aldehyde dehydrogenase activities in rats treated with m-xylene 8, 12 or 16 times were significantly decreased compared to those pretreated one or four times. In the early stage of xylene administration, proliferation of SERs were seen whereas SERs were decreased and RERs were clearly increased in xylene-treated rats 16 times. These results indicate that the frequency of xylene injection may influence upon the changes in xylene metabolite, m-methylhippuric acid and it may be due to induction of xylene metabolizing enzymes.

흰쥐에게 Ethanol과 Xylene 병행 투여시 간손상에 미치는 영향

윤인숙(In-Sook Yoon),이혜자(Hye-Ja Lee) 한국콘텐츠학회 2010 한국콘텐츠학회논문지 Vol.10 No.7

흰쥐에게 ethanol과 xylene을 투여시 간손상 정도를 알아보기 위해 정상적인 S-D계 수컷 흰쥐를 4군으로 나누어 실험을 수행하였다. 생화학적 검사 결과에서 혈청 AST, ALT, LDH활성은 대조군보다 ethanol 투여군, xylene투여군, ethanol+xylene투여군으로 갈수록 활성이 유의하게 증가되었으며, 그 증가율은 ethanol+xylene투여군에서 가장 높게 나타났다. 산화적 손상정도를 알아보기 위한 혈청 MDA함량은 대조군보다 ethanol투여군과 xylene투여군 그리고 ethanol+xylene투여군으로 갈수록 활성이 증가되었으며, 그 증가율은 ethanol+xylene투여군에서 가장 높게 나타났다. 그러나 혈청 ALP는 대조군에 비해 모두 감소하였다. Xylene의 투여시 최종대사산물인 요 중 m-methylhippuric acid는 대조군에 비해 xylene투여군과 ethanol+xylene투여군에서 증가되었으며, 그 증가율은 xylene투여군이 ethanol+xylene투여군보다 더 높게 나타났다. 간 조직검사에서는 xylene투여군은 지방변화를, ethanol+xylene 투여군은 경도~중등도의 괴사와 염증을 보였다. 이상의 혈청학적 검사와 조직학적 검사결과 xylene 단독 투여에 비하여 ethanol과 xylene을 병행 투여했을 때 간손상이 더 크게 초래된 것으로 나타났다. To evaluate an effect of liver damage on ethanol and xylene exposure, experiments on normal male rats of the S-D strain were performed in 4 groups. The biochemical results suggest that the ethanol group had significantly higher levels of AST, ALT, LDH and also, the xylene group had notably higher levels of AST, ALT, LDH along with MMHPA than those of the control groups. The levels of AST, ALT and LDH in the ethanol+xylene group were drastically higher than those in the control, ethanol and xylene groups. But, there were significantly lower ALP levels in the xylene and ethanol+xylene groups than both the control and ethanol groups. The histological features of rat livers treated with alcohol, or xylene proved to be normal. But the rat livers treated with ethanol+xylene showed mild to moderate necrosis and inflammation as well as minimal fatty changes. The results in this experiment suggest that liver functions decreased when medicated together with xylene and ethanol rather than solely with xylene.

흰쥐 성장기간에 따른 Xylene의 독성에 관한 연구

이혜자,이상희,전태원,이상일,윤종국 대한의생명과학회 2000 Journal of biomedical laboratory sciences Vol.6 No.3

실험동물에 있어서 연령 차이에 따라서 xylene 독성이 어떠한 차이가 있는지를 검토하는 일환으로 5주령 및 12주령 흰쥐에 50% m-xylene을 체중 100 g 당 0.25 ml씩 1회 투여한 다음 24시간 후에 처치하여 다음과 같은 결과를 얻었다. Xylene 투여로 인한 요 중 methylhippuric acid 함량은 5주령군이 12주령군에 비해 현저하게 높게 나타났다. 그리고 간조직 중 cytochroms P-450 함량은 대조군에 있어서 5주령군이 12주령군 보다 약 50% 정도 낮게 나타났으나 xylene 투여로 인한 cytochrome P-450 함량 증가율은 12주령군 보다 5주령군에서 높게 나타났다. 간 alcohol dehydrogenase 활성치도 대조군에 있어서는 5주령군이 12주령 보다 약 35% 정도 낮게 나타났으나 xylene 투여로 인한 본 효소의 활성 증가율은 5주령군에서 오히려 높게 나타났다. 그러나 간 aldehyde dehydrogenase 활성치는 대조군 및 xylene 투여군 모두 5주령과 12주령간에 유의한 차이를 나타내지 않았다. 한편 xylene 투여시 체중 당 간무게, 간조직 malondialdehyde 함량 및 혈청 ALT 활성 변동을 통하여 간손상 정도를 상호 비교 관찰하였을 때, 12주령군이 5주령 실험동물 보다 간손상이 다소 심하게 나타남을 알 수가 있었다. 이상 실험결과는 연령에 따라 xylene에 의한 간손상의 차이는 이물질의 생체내 대사율이 달리 나타나기 때문일 것으로 생각된다. To evaluate an effect of aging on the xylene toxicity, 50% m-xylene in olive oil (0.25 ml/100 g body wt.) was administered to 5 week and 12 week-old rats one times intraperitoneally and sacrificed at 24 hrs afterwards. The increasing rate of urinary H-methylhippuric acid concentration was higher in 12 week-old rats than 5 week-old rats by the treatment of m-xylene. On the liver function findings, i.e., liver weight/body weight (%), serum levels of ALT activity and hepatic malondialdehyde content, 12 week-old rats showed more severe liver injury than 5 weeks those in xylene-treated rats. And the hepatic cytochrome P-450 contents was higher in 12 weeks rats than those of in 5 week-old rats, but the increasing rate of that was lower in 12 week-old rats. Hepatic alcohol dehydrogenase activity was also higher in 12 week-old rats than in 5 week-old rats whereas the increasing rate of that was higher in 5 week-old than those in 12 week-old rats by the xylene treatment. Furthermore, the hepatic aldehyde dehydrogenase activities were no differences between the 5 and 12 week-old rats both in the control and xylene-treated group, In conclusion, age may influences upon the hepatotoxicity with xylene and it may be responsible for xylene metabolism in rats.

흰쥐에 있어서 사염화탄소에 의한 간손상이 Xylene 대사에 미치는 영향

이혜자(Hye Ja Lee),조현국(Hyun Gug Cho),윤종국(Chong Guk Yoon) 한국환경보건학회 1999 한국환경보건학회지 Vol.25 No.1

흰쥐에 있어서 CCl_4에 의한 간손상이 xylene 대사에 어떠한 영향을 미치는지를 알아보기 위하여 실험동물에 CCl_4 투여하며 간손상을 유도한 다음, m-xylene을 투여한 후 24시간 동안 요 중 m-methylhippuric acid 함량을 측정한 결과 m-xylene만 투여한 실험군 보다 요중 m-methylhippuric acid함량이 유의하게 감소되었다. 이때 xylene 대사에 관여하는 cytochrome P-450 type aniline hydroxylase 및 aldehyde dehydrogenase 활성 역시 간손상이 유도된 실험동물에서 유의한 감소를 보였다. 또한 CCl_4 의하여 간손상을 유도한 다음 xylene을 투여한 실험동물의 간세포 미세구조의 소포체 변화와 xylene 대사율의 저하는 xylene 대사에 관여하는 효소 단백 합성과 관련되어 나타난 결과로 생각된다. To evaluate an effect of pathological liver damage on the xylene metabolism, the rats were induced acute liver damage with 4 times of CCl_4 injection (0.1 ㎖ of 50% in olive oil/100 g body wt.) every other days. The injection of one dose xylene (0.25 ㎖ of 50% in olive oil/100 g body wt.) showed the significantly decreased urinary methylhippuric acid concentration throughout 24 hr in the liver damaged-animals than the normal group treated with only xylene. The activities of hepatic microsomal aniline hydroxylase and aldehyde dehydrogenase were significantly decreased in CCl_4 pretreated rats than the normal group at 24 hr after injection of m-xylene. Furthermore, the more severe ultrastructural changes were revealed in the hepatic cell of liver damaged-animals than in that of normal group by the injection of one dose xylene. These results suggested that the rats induced liver damage with CCl_4 may reduce the xylene metabolism.

Xylene 대사 효소 활성에 미치는 주.야 시차의 영향

이혜자,윤종국 한국환경보건학회 2001 한국환경보건학회지 Vol.27 No.2

To evaluate an effect of circadian variation on the xylene metabolizing enzyme activities, 50% m-xylene in olive oil(0.25 mℓ/100g body weight) was intraperitoneally administered to the rats every other day for 6 days both in the night; 24:00 and the day; 12:00. Then animals were sacrigiced at 8hr after last injection of m-xylene. Hepatic microsomal cytochrome p450 contents were more increased both in control and xylene treated rats of night phase than those of day phase. But the activity of hepatic alcohol dehydrogenase(ADH) in control of night phase showed the similar value with that in those of day phase and xylene treated rats of day phase showed an increasing tendency of hepatic ADH activity as those of night phase showing similar activity. Furthermore, control rats of night phase than those of day phase. And by xylene treatment, enzyme activities of rats of day phase were higher tendency in rats of control but those of night phase were somewhat inhibited. Besides, xylene-treated animals of night phase showed increasing tendency of urinary methylhippuric acid concentration compared with those of day phase. On the other hand, liver weight per body weight(%), hepatic lipid peroxide content and serum xanthine oxidase activity were higher in night phase. And the activities of hepatic oxygen free radical metabolizing enzymes such as xanthine oxidase, gluthathione S-transferase, and xylene-treated rats of night phase than those of day phase. In conclusion, it can be hypothesized on the basis of the results that the accumulation rate of m-xylene intermediate metabolite, i.e. m-methylbenzaldehyde in liver tissus may be higher in night phase than in day phase and it may be responsible for higher liver toxicity in bight phase than in day phase.

토양 서식 미생물을 이용한 자일렌(xylene) 분해특성 조사

최필권(Phil Kweon Choi),허평(Pyeung Heo),이상섭(Sang Seob Lee) 大韓環境工學會 2013 대한환경공학회지 Vol.35 No.6

본 연구의 목적은 유류로 오염된 토양으로부터 분리된 고효율 분해균주에 의한 xylene 분해특성을 조사하고 분해과정을 밝히는 것이다. P. putida BJ10에 의해 mineral salts medium (MSM)배지내에서 24시간 배양 후 제거율은 o, m, p-xylene각각 94, 90, 98%였으며 3% 이하의 대조군과 명확한 차이를 보였다. 또한 유류로 오염된 토양내에서의 9일 경과 후 xylene제거율은 유류 분해균주 주입구에서 66%였으며 무처리 일반토양에서 32%, 멸균 토양에서 8%로서 P. putida BJ10에 의한 유효성을 확인할 수 있었다. 또한 시간경과에 따른 대사산물을 분석한 결과 o-xylene의 분해 경로에서는 6시간 경과 후 3-methylcatechol, 24시간 경과 후 o-toluic acid 가 분해산물로서 검출되어 최종산물로서의 o-toluic acid 가 확인되었으며 중간산물의 변환과정은 기존 발표된 연구들과 다소 다른 결과를 나타내었다. The purpose of this study is to investigate the biodegradation characteristics of the xylene by BTEX-degrading bacteria, Pseudomonas putida BJ10, isolated from oil-contaminated soil and bio-degradation pathway of the xylene. The removal efficiencies of o, m, p-xylene in mineral salts medium (MSM) by P. putida BJ10 were 94, 90 and 98%, respectively for 24 hours. It shows clear difference compared with the control groups which were below 3%. The removal efficiencies of BTEX by P. putida BJ10 in gasoline- contaminated soil were 66% for 9 days. They were clearly distinguished from the control groups (control and sterilized soil) which were 32 and 8%. 3-methylcatechol and o-toluic acid were detected after 6 and 24 hours during the o-xylene biodegradation pathway. Therefore, we confirmed o-toluic acid as the final metabolite. And intermediate-products were somewhat different with previously published studies of the transformation pathway from o-xylene to 3-methylcatechol.

Urine Methyl Hippuric Acid Levels in Acute Pesticide Poisoning: Estimation of Ingested Xylene Volume and Association with Clinical Outcome Parameters

Choi, Chi-Young,Cho, NamJun,Park, Su-Yeon,Park, Samel,Gil, Hyo Wook,Hong, Sae-Yong The Korean Academy of Medical Sciences 2017 JOURNAL OF KOREAN MEDICAL SCIENCE Vol.32 No.12

<P>To determine the relationship between the oral ingestion volume of xylene and methyl hippuric acid (MHA) in urine, we measured MHA in 11 patients whose ingested xylene volume was identified. The best-fit equation between urine MHA and ingested amount of xylene was as follows: y (ingested amount of xylene, mL/kg) = −0.052x<SUP>2</SUP> + 0.756x (x = MHA in urine in g/g creatinine). From this equation, we estimated the ingested xylene volume in 194 patients who had ingested pesticide of which the formulation was not available. Our results demonstrated that oxadiazole, dinitroaniline, chloroacetamide, organophosphate, and pyrethroid were xylene-containing pesticide classes, while the paraquat, glyphosate, glufosinate, synthetic auxin, fungicide, neonicotinoid, and carbamate classes were xylene-free pesticides. Sub-group univariate analysis showed a significant association between MHA levels in urine and ventilator necessity in the pyrethroid group. However, this association was not observed in the organophosphate group. Our results suggest that MHA in urine is a surrogate marker for xylene ingestion, and high urine MHA levels may be a risk factor for poor clinical outcome with some pesticide poisoning.</P>