Suppression of growth defects of α-amylase secreting Escherichia coli by signal sequence fusion
(1988) In FEMS Microbiology Letters 55(1). p.3-7- Abstract
Two fusions of the Bacillus stearothermophilus α-amylase gene (amyS) with lacpoZ′ were constructed. The first, being a transcriptional fusion, placed amyS directly under lac promoter control eliminating interference by the endogenous promoter. IPTG induction of amyS transcription in this construction resulted in liberation of periplasmic proteins and eventually cell lysis. The other fusion replaced eight N-terminal amino acid residues from the signal sequence by 11 residues from the lacZ′ moiety in pUC-18. Translation initiation signals were also replaced by those from lacpoZ′. In this case IPTG induction resulted in secretion of approx. 35% of total α-amylase activity in the growth medium after 24 h growth without detectable cell... (More)
Two fusions of the Bacillus stearothermophilus α-amylase gene (amyS) with lacpoZ′ were constructed. The first, being a transcriptional fusion, placed amyS directly under lac promoter control eliminating interference by the endogenous promoter. IPTG induction of amyS transcription in this construction resulted in liberation of periplasmic proteins and eventually cell lysis. The other fusion replaced eight N-terminal amino acid residues from the signal sequence by 11 residues from the lacZ′ moiety in pUC-18. Translation initiation signals were also replaced by those from lacpoZ′. In this case IPTG induction resulted in secretion of approx. 35% of total α-amylase activity in the growth medium after 24 h growth without detectable cell lysis.
(Less)
- author
- Suominen, Ilari ; Käpylä, Jarmo ; Tilgmann, Carola LU ; Glumoff, Virpi and Mäntsälä, Pekka
- publishing date
- 1988
- type
- Contribution to journal
- publication status
- published
- keywords
- Bacillus stearothermophilus, Escherichia coli, Periplasmic protein, Protein secretion, α-amylase
- in
- FEMS Microbiology Letters
- volume
- 55
- issue
- 1
- pages
- 5 pages
- publisher
- Oxford University Press
- external identifiers
-
- scopus:0023698669
- ISSN
- 0378-1097
- language
- English
- LU publication?
- no
- id
- fa6c734f-1c9a-4ffa-9ea2-51db1c17c6b1
- date added to LUP
- 2016-04-11 13:26:57
- date last changed
- 2021-01-03 06:09:54
@article{fa6c734f-1c9a-4ffa-9ea2-51db1c17c6b1, abstract = {{<p>Two fusions of the Bacillus stearothermophilus α-amylase gene (amyS) with lacpoZ′ were constructed. The first, being a transcriptional fusion, placed amyS directly under lac promoter control eliminating interference by the endogenous promoter. IPTG induction of amyS transcription in this construction resulted in liberation of periplasmic proteins and eventually cell lysis. The other fusion replaced eight N-terminal amino acid residues from the signal sequence by 11 residues from the lacZ′ moiety in pUC-18. Translation initiation signals were also replaced by those from lacpoZ′. In this case IPTG induction resulted in secretion of approx. 35% of total α-amylase activity in the growth medium after 24 h growth without detectable cell lysis.</p>}}, author = {{Suominen, Ilari and Käpylä, Jarmo and Tilgmann, Carola and Glumoff, Virpi and Mäntsälä, Pekka}}, issn = {{0378-1097}}, keywords = {{Bacillus stearothermophilus; Escherichia coli; Periplasmic protein; Protein secretion; α-amylase}}, language = {{eng}}, number = {{1}}, pages = {{3--7}}, publisher = {{Oxford University Press}}, series = {{FEMS Microbiology Letters}}, title = {{Suppression of growth defects of α-amylase secreting Escherichia coli by signal sequence fusion}}, volume = {{55}}, year = {{1988}}, }