Nucleic acid sequences and expression systems for heparinase II

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Lyase

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536 232, C12N 988, C12N 1560

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active

056817339

ABSTRACT:
The present invention describes the isolation and sequence of genes from Flavobacterium heparinum encoding heparin and heparan sulfate degrading enzymes, heparinase II and heparinase III (EC 4.2.2.8). It further describes a method of expressing and an expression for heparinases I, II and III using a modified ribosome binding region derived from a promoter from glycosaminoglycan lyase genes of F. heparinum. Also, a multi-step protein purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography and hydroxylapatite chromatography is outlined. Antibodies against a post-translational modification moiety common to Flavobacterium heparinum proteins and a method to obtain antibodies specific to these moieties and to the amino acid sequences of heparinases I, II and III are described.

REFERENCES:
patent: 5389539 (1995-02-01), Sasisekharan et al.
Lohse, et al., The Journal of Biological Chemistry, vol. 267:34, pp. 24347-24335 (1992).
Sasisekharan, et al., Proc. Natl. Acad. Sci., USA, vol. 90, pp. 3660-3664 (1993).
Zimmermann, et al., Applied and Environmental Microbiology, vol. 56:11, pp. 2593-3594 (1990).

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