Control of Formation of Active Soluble or Inactive Insoluble Baker's Yeast Alpha-glucosidase PI in Escherichia Coli by Induction and Growth Conditions

Overview

Journal Mol Gen Genet

Specialties Genetics
Molecular Biology

Date 1989 Mar 1

PMID 2659969

Citations 16

Authors

E Kopetzki

G Schumacher

P Buckel

Affiliations

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Abstract

Using standard growth conditions (LB medium, 37 degrees C, induction with 5 mM IPTG) yeast alpha-glucosidase PI expressed under the control of the regulated tac-hybrid promoter results in the synthesis of insoluble aggregated alpha-glucosidase granules in Escherichia coli. Under these conditions active soluble alpha-glucosidase amounts to less than 1% of the heterologously produced protein. However, the amount of soluble active alpha-glucosidase was dramatically increased when the strong tac-hybrid promoter was to a limited extent induced. This was achieved at concentrations of 0.01 mM IPTG or of 1% lactose or lower in a lactose-permease deficient host strain containing the lacIq repressor gene on an R-plasmid. The formation of active soluble alpha-glucosidase was almost 100% when E. coli cells induced in this manner were cultivated under conditions that reduced growth rate, i.e. at decreased temperature, extreme pH values or in minimal and complete media supplemented with different carbon sources.

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