• 제목/요약/키워드: bacteroides stercoris HJ-15

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사람 장내세균군집 유래 Bacteorides stericoris HJ-15의 Chondroitinase ABC의 클로닝 (Cloning of Chondroitinase ABC from Bacteroides stercoris HJ-15, a Human Intestinal Anaerobic Bacterium)

  • 방서현;심주원;현양진;김동현
    • 한국미생물·생명공학회지
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    • 제44권2호
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    • pp.140-144
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    • 2016

  • 사람의 분변으로부터 분리한 Bacteroides stericoris HJ-15로부터 chondroitinase ABC 유전자를 클로닝하였다. 클로닝한 chondroitinase ABC 유전자는 3,090 bp, 1,029 아미노산으로 구성되어 있었다. B. stercoris chondroitinase ABC 유전자는 이미 보고된 chondroitinase ABC 유전자들과 호몰로지가 없었으나, 아미노산서열에서는 82% 호몰로지를 보였다. T7 promoter를 가진 pET-26b+ expression vector에 클로닝한 chondroitinase ABC 유전자를 Escherichia coli BL21 (DE3)에서 발현하여 정제한 재조합 chondroitinase ABC는 chondroitin sulfate A, B 및 C를 모두 분해하였다.

  • https://doi.org/10.4014/mbl.1512.12007 인용 PDF KSCI KPUBS

Degradation of Acharan Sulfate and Heparin by Bacteroides stercoris HJ-15, a Human Intestinal Bacterium

  • Kim, Dong-Hyun;Kim, Byung-Taek;Park, Sun-Yong;Kim, Na-Young;Han, Myung-Joo;Shin, Kuk-Hyun;Kim, Wan-Suk;Kim, Yeong-Sik
    • Archives of Pharmacal Research
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    • 제21권5호
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    • pp.576-580
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    • 1998

  • When glycosaminoglycan (GAG)-degrating enzymes were measured in normal human stool suspensions, all 5 tested different stools degraded titrable heparin and acharan sulfate. GAG-degrading bacteria were screened from the isolates of human stools. Among them, HJ-15 had the most potent activities of heparinases (GAGs-degrading enzymes). However, HJ-15 produced the enzyme even if in the media without heparin. Acharan sulfate lyase was induced by acharan sulfate and heparin. Heparinase production was also induced by these GAGs. These enzymes, acharan sulfate lyase and heparinase, were produced in exponential and stationary phase of HJ-15 growth, respectively. Optimal pHs of the acharan sulfate lyase and heparinase activities were 7.2 and 7.5 respectively. the biochemical properties of HJ-15 was similar to those of B. stercoris. However, difference from B. stercoris was utilization of raffinose. this HJ-15 also degraded chondroitin sulfates A and C.

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Purification and Characterization of Heparin Lyase I from Bacteroides stercoris HJ-15

  • Kim, Wan-Seok;Kim, Byung-Taek;Kim, Dong-Hyun;Kim, Yeong-Shik
    • BMB Reports
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    • 제37권6호
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    • pp.684-690
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    • 2004

  • Heparin lyase I was purified to homogeneity from Bacteroides stercoris HJ-15 isolated from human intestine, by a combination of DEAE-Sepharose, gel-filtration, hydroxyapatite, and CM-Sephadex C-50 column chromatography. This enzyme preferred heparin to heparan sulfate, but was inactive at cleaving acharan sulfate. The apparent molecular mass of heparin lyase I was estimated as 48,000 daltons by SDS-PAGE and its isoelectric point was determined as 9.0 by IEF. The purified enzyme required 500 mM NaCl in the reaction mixture for maximal activity and the optimal activity was obtained at pH 7.0 and $50^{\circ}C$. It was rather stable within the range of 25 to $50^{\circ}C$ but lost activity rapidly above $50^{\circ}C$. The enzyme was activated by $Co^{2+}$ or EDTA and stabilized by dithiothreitol. The kinetic constants, $K_m$ and $V_{max}$ for heparin were $1.3{\times}10^{-5}\;M$ and $8.8\;{\mu}mol/min{\cdot}mg$. The purified heparin lyase I was an eliminase that acted best on porcine intestinal heparin, and to a lesser extent on porcine intestinal mucosa heparan sulfate. It was inactive in the cleavage of N-desulfated heparin and acharan sulfate. In conclusion, heparin lyase I from Bacteroides stercoris was specific to heparin rather than heparan sulfate and its biochemical properties showed a substrate specificity similar to that of Flavobacterial heparin lyase I.

  • https://doi.org/10.5483/BMBRep.2004.37.6.684 인용 PDF