Heterologous expression and secretion of Lactobacillus amylovorus alpha-amylase in Leuconostoc citreum.
To develop a gene expression system for Leuconostoc genus, construction of expression vector and expression of a heterologus protein in Leuconostoc was performed. Alpha-amylase gene from Lactobacillus amylovorus was cloned into a Leuconostoc cloning vector, pLeuCM, with its own signal peptide. pLeuCMamy was introduced into Leuconostoc citreum CB2567 and a successful expression of alpha-amy gene was confirmed by enzyme activity assays. About 90% of alpha-amylase activity was detected in the culture broth, revealing most of expressed alpha-amylase was secreted out cells. The signal sequence of alpha-amy gene is a good candidate for the secretion of heterologous protein by using Leuconostoc host-vector system.
Eom
Hyun-Ju
HJ
Department of Food Science and Technology, Research Center for Bioresource and Health, Chungbuk National University, Cheongju, 361-763, South Korea.
Moon
Jin-Seok
JS
Seo
Eun-Young
EY
Han
Nam Soo
NS
D003001:Cloning, Molecular; D005798:Genes, Bacterial; D007779:Lactobacillus acidophilus / Q000201:enzymology* / Q000235:genetics; D007934:Leuconostoc / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism*
D000516:alpha-Amylases