Article (Scientific journals)
Overproduction and properties of the mannuronate alginate lyase AlxMB
Malissard, Martine; Chavagnat, Frédéric; Duez, Colette et al.
1995In FEMS Microbiology Letters, 126 (2), p. 105-111
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Keywords :
amino acid sequence; base sequence; cloning, molecular; escherichia coli/enzymology/genetics; molecular sequence data; plasmids/genetics; polysaccharide-lyases/*biosynthesis/chemistry/genetics/isolation; purification
Abstract :
[en] 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 mu g 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 ALxM(A) and unnicked ALxM(B) alginate lyases have identical alginate-degrading activities at high salt concentrations.
Research center :
CIP - Centre d'Ingénierie des Protéines - ULiège
Disciplines :
Microbiology
Biochemistry, biophysics & molecular biology
Author, co-author :
Malissard, Martine;  Université Claude Bernard - Lyon 1 - UCLB > Laboratoire de Biochimie Microbienne
Chavagnat, Frédéric;  Université Claude Bernard - Lyon 1 - UCLB > Laboratoire de Biochimie Microbienne
Duez, Colette ;  Université de Liège - ULiège > Centre d'ingénierie des protéines
Vacheron, Marie-Jeanne;  Université Claude Bernard - Lyon 1 - UCLB > Laboratoire de Biochimie Microbienne
Guinand, Micheline;  Université Claude Bernard - Lyon 1 - UCLB > Laboratoire de Biochimie Microbienne
Michel, Georges;  Université Claude Bernard - Lyon 1 - UCLB > Laboratoire de Biochimie Microbienne
Ghuysen, Jean-Marie ;  Université de Liège - ULiège > Centre d'ingénierie des protéines
Language :
English
Title :
Overproduction and properties of the mannuronate alginate lyase AlxMB
Publication date :
15 February 1995
Journal title :
FEMS Microbiology Letters
ISSN :
0378-1097
eISSN :
1574-6968
Publisher :
Blackwell Publishing, Oxford, United Kingdom
Volume :
126
Issue :
2
Pages :
105-111
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
FRSM - Fonds de la Recherche Scientifique Médicale [BE]
Available on ORBi :
since 26 May 2011

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