Bacterial expression of an active class Ib chitinase fromCastanea sativacotyledons
✍ Scribed by Isabel Allona; Carmen Collada; Rosa Casado; Javier Paz-Ares; Cipriano Aragoncillo
- Book ID
- 104619248
- Publisher
- Springer
- Year
- 1996
- Tongue
- English
- Weight
- 595 KB
- Volume
- 32
- Category
- Article
- ISSN
- 0167-4412
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✦ Synopsis
Ch3, an endochitinase of 32 kDa present in Castanea sativa cotyledons, showed in vitro antifungal properties when assayed against Trichoderma viride. The characterization of a cDNA clone corresponding to this protein indicated that Ch3 is a class Ib endochitinase that is synthesized as a preprotein with a signal sequence preceding the mature polypeptide. Bacterial expression of mature Ch3 fused to the leader peptide of the periplasmic protein ompT resulted in active Ch3 enzyme. A plate assay was adapted for semi-quantitative determination of chitinase activity secreted from cultured bacteria, which should facilitate the identification of mutants with altered capacity to hydrolyse chitin.