Production of pectin methylesterase from Erwinia chrysanthemi B374 in Bacillus subtilis.
The gene coding for pectin methylesterase (PME) of Erwinia chrysanthemi B374 (pme) was cloned by a polymerase chain reaction. The pme gene was expressed in Bacillus subtilis using a secretion vector based on the promoter and signal sequence of the alpha-amylase gene from B. amyloliquefaciens. The cultivation of B. subtilis cells carrying the cloned pme resulted in efficient secretion of PME into the culture medium based on enzymatic and sodium dodecyl sulphate-poly-acrylamide gel electrophoresis characterizations. The NH2-terminal sequence analysis of the secreted PME revealed two different NH2-termini. Heterologous processing was probably due to a second putative signal peptidase cleavage site at the joint region between the PME and alpha-amylase signal peptide.
Heikinheimo
R
R
Genesit Ltd., Helsinki, Finland.
Hemilä
H
H
Pakkanen
R
R
Palva
I
I
D001412:Bacillus subtilis / Q000235:genetics; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D003470:Culture Media; D004885:Erwinia / Q000201:enzymology*; D005285:Fermentation; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
D003470:Culture Media; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase