ArticleApplied and environmental microbiology2005
Kluyveromyces lactis LAC4 promoter variants that lack function in bacteria but retain full function in K. lactis.
Article in Applied and environmental microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 64 citations in OpenAlex.
- Creating a large designer cellulosome in yeast to boost ethanol production.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- A convenient broad-host counterselectable system endowing rapid genetic manipulations of Kluyveromyces lactis and other yeast species.Microbial cell factories · 2024Article
- Directed evolution for soluble and active periplasmic expression of bovine enterokinase in Escherichia coli.Scientific reports · 2022Article
- Constructing a yeast to express the largest cellulosome complex on the cell surface.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- A Novel Regulated Hybrid Promoter That Permits Autoinduction of Heterologous Protein Expression in Kluyveromyces lactis.Applied and environmental microbiology · 2019Article
- Valuation of agro-industrial wastes as substrates for heterologous production of α-galactosidase.Microbial cell factories · 2018Article
- Comparison of Yeasts as Hosts for Recombinant Protein Production.Microorganisms · 2018Review
- Article
- Genome and metabolic engineering in non-conventional yeasts: Current advances and applications.Synthetic and systems biotechnology · 2017Review
- Heterologous expression of an α-amylase inhibitor from common bean (Phaseolus vulgaris) in Kluyveromyces lactis and Saccharomyces cerevisiae.Microbial cell factories · 2017Article
- Yeast Kluyveromyces lactis as host for expression of the bacterial lipase: cloning and adaptation of the new lipase gene from Serratia sp.Journal of industrial microbiology & biotechnology · 2015Article
- Use of synthetic genes for cloning, production and functional expression of the bacteriocins enterocin A and bacteriocin E 50-52 by Pichia pastoris and Kluyveromyces lactis.Molecular biotechnology · 2014Article
- Construction of a Kluyveromyces lactis ku80⁻ host strain for recombinant protein production: extracellular secretion of pectin lyase and a streptavidin-pectin lyase chimera.Molecular biotechnology · 2014Article
- Carbon source dependent promoters in yeasts.Microbial cell factories · 2014Review
- Cloning, production, and functional expression of the bacteriocin sakacin A (SakA) and two SakA-derived chimeras in lactic acid bacteria (LAB) and the yeasts Pichia pastoris and Kluyveromyces lactis.Journal of industrial microbiology & biotechnology · 2013Article
- Metabolic engineering of Kluyveromyces lactis for L-ascorbic acid (vitamin C) biosynthesis.Microbial cell factories · 2013Article
- Uptake of radiolabeled GlcNAc into Saccharomyces cerevisiae via native hexose transporters and its in vivo incorporation into GPI precursors in cells expressing heterologous GlcNAc kinase.FEMS yeast research · 2012Article
- How recombinant swollenin from Kluyveromyces lactis affects cellulosic substrates and accelerates their hydrolysis.Biotechnology for biofuels · 2011Article
- A set of aspartyl protease-deficient strains for improved expression of heterologous proteins in Kluyveromyces lactis.FEMS yeast research · 2011Article
- Production of isotopically labeled heterologous proteins in non-E. coli prokaryotic and eukaryotic cells.Journal of biomolecular NMR · 2010Article
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2 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
The strong LAC4 promoter (P(LAC4)) from Kluyveromyces lactis has been extensively used to drive expression of heterologous proteins in this industrially important yeast. A drawback of this expression method is the serendipitous ability of P(LAC4) to promote gene expression in Escherichia coli. This can interfere with the process of assembling expression constructs in E. coli cells prior to their introduction into yeast cells, especially if the cloned gene encodes a protein that is detrimental to bacteria. In this study, we created a series of P(LAC4) variants by targeted mutagenesis of three DNA sequences (PBI, PBII, and PBIII) that resemble the E. coli Pribnow box element of bacterial promoters and that reside immediately upstream of two E. coli transcription initiation sites associated with P(LAC4). Mutation of PBI reduced the bacterial expression of a reporter protein (green fluorescent protein [GFP]) by approximately 87%, whereas mutation of PBII and PBIII had little effect on GFP expression. Deletion of all three sequences completely eliminated GFP expression. Additionally, each promoter variant expressed human serum albumin in K. lactis cells to levels comparable to wild-type P(LAC4). We created a novel integrative expression vector (pKLAC1) containing the P(LAC4) variant lacking PBI and used it to successfully clone and express the catalytic subunit of bovine enterokinase, a protease that has historically been problematic in E. coli cells. The pKLAC1 vector should aid in the cloning of other potentially toxic genes in E. coli prior to their expression in K. lactis.
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