Overproduction and properties of the mannuronate alginate lyase AlxMB.

نویسندگان

  • M Malissard
  • F Chavagnat
  • C Duez
  • M J Vacheron
  • M Guinand
  • G Michel
  • J M Ghuysen
چکیده

In previous studies (Malissard et al., FEMS Microbiol. Lett. (1993) 110, 101-106), the alginate lyase AlxM of the marine bacterium ATCC 433367 was produced in Escherichia coli TC4/pAL-A3 with a yield of 50 micrograms per litre of culture. The polypeptide chain was cleaved between two cysteine residues, C169 and C183, themselves linked by a disulphide bridge. AlxM has now been overproduced in E. coli BL21(DE3)/pAL-Sur/pLysS. Under conditions in which formation of inclusion bodies can be avoided, the enzyme is synthesized as a catalytically active, water-soluble, unnicked polypeptide with a yield of 32 mg per litre of culture. It has been purified to protein homogeneity using a one-step procedure. The nicked AlxMA and unnicked AlxMB alginate lyases have identical alginate-degrading activities at high salt concentrations.

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عنوان ژورنال:
  • FEMS microbiology letters

دوره 126 2  شماره 

صفحات  -

تاریخ انتشار 1995