• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.140-144
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• 2016

The gene encoding chondroitinase ABC from a genomic library of Bacteroides stercoris HJ-15, which was isolated from human feces, was cloned. The cloned gene consisted of 3,090 bp and was predicted to encode a 1,029−amino-acid protein. The B. stercoris chondroitinase ABC gene was not homologous to other chondroitinase ABC genes; however, its amino acid sequence showed 71% homology to that of Bacteroides thetaiotaomicron. The gene was cloned in the pET-26b+ expression vector and expressed under the T7 promoter in Escherichia coli BL21(DE3). The purified recombinant chondroitinase ABC degraded chondroitin sulfates A, B, and C.

• Archives of Pharmacal Research
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• v.21 no.5
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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.

• BMB Reports
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• v.37 no.6
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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.

• Proceedings of the PSK Conference
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• 2002.04a
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• pp.93-93
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• 2002

• Proceedings of the PSK Conference
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• 2000.10a
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• pp.203.1-203.1
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• 2000

• Proceedings of the PSK Conference
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• 2001.04a
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• pp.178.3-179
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• 2001

• Proceedings of the PSK Conference
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• 2000.04a
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• pp.154.3-155
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• 2000