1887

Abstract

Summary: Strain N106, a newly isolated soil actinomycete classified in the genus on the basis of its chemotaxonomy, produced an extracellular chitosanase and was highly active in chitosan degradation. A gene library of sp. N106 was constructed in the shuttle vector pFD666 and recombinant plasmids carrying the chitosanase gene () were identified using the 5′-terminal portion of the chitosanase gene from sp. N174 as a hybridization probe. One plasmid, pCSN106-2, was used to transform TK24. The chitosanase produced by (pCSN106-2) is a protein of 29.5 kDa, with a pl 8.1, and hydrolyses chitosan by an endo-mechanism giving a mixture of dimers and trimers as end-products. N-terminal sequencing revealed that the mature chitosanase is a mixture of two enzyme forms differing by one N-terminal amino acid. The gene is 79.5% homologous to the gene from sp. N174. At the amino acid level, both chitosanases are homologous at 74.4% and hydrophobic cluster analysis revealed a strict conservation of structural features. This chitosanase is the third known member of family 46 of glycosyl hydrolases.

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1995-10-01
2021-10-18
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