Premium
Extracellular Release of Recombinant α‐Amylase from Escherichia coli Using Pulsed Electric Field
Author(s) -
Shiina Satoshi,
Ohshima Takayuki,
Sato Masayuki
Publication year - 2004
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp049760u
Subject(s) - periplasmic space , recombinant dna , escherichia coli , extracellular , amylase , intracellular , chemistry , biochemistry , enzyme , biology , microbiology and biotechnology , gene
It is difficult for Escherichia coli to secrete products such as recombinant enzymes, because the Gram‐negative bacterium has a double membrane structure and so some of the products are accumulated in a periplasmic space. In this study, we demonstrated that recombinant α‐amylase can be released from recombinant E. coli HB101/pHI301A during cultivation by applying a pulsed electric field (PEF). When a PEF (12 kV, 2 Hz) was applied for 30 min with an interval of 30 min from the point of OD 660 = 0.7, the amount of released α‐amylase was about 30% of the total amount of α‐amylase produced in the cells. As a result of SDS‐PAGE and activity staining analyses, it was confirmed that the released proteins were not all of the intracellular proteins, and the α‐amylase, which was identical with intracellular α‐amylase, was released by applied PEF cultivation. PEF treatment could be useful for easy release of periplasmic protein with selectivity.