1887

Abstract

Two types of Tn-induced mutants defective in extracellular amylase activity were isolated from the moderate halophile DSM 5425. Type I mutants displayed amylase activity in the periplasm, and were unable to use any of the carbon sources tested, including starch and its hydrolysis product maltose. The type II mutant was affected in the gene responsible for the synthesis of the extracellular α-amylase. This gene () was isolated by functional complementation of mutant II and sequenced. The deduced protein (AmyH) showed a high degree of homology to a proposed family of α-amylases consisting of enzymes from (), , streptomycetes, insects and mammals. AmyH contained the four highly conserved regions in amylases, as well as a high content of acidic amino acids. The gene was functional in the moderate halophile and, when cloned in a multicopy vector, in . AmyH is believed to be the first extracellular-amylase-encoding gene isolated from a moderate halophile, a group of extremophiles of great biotechnological potential. In addition, and were able to secrete the thermostable α-amylase from , indicating that members of the genus are good candidates for use as cell factories to produce heterologous extracellular enzymes.

Keyword(s): Halomonas , halophile and α-amylase
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2000-04-01
2019-10-14
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