Abstract:
:Expression of recombinant proteins in Escherichia coli often requires use of inducible promoters to shorten the lag phase and improve protein productivity and final protein titer. Synthetic molecules that cannot be metabolized by E. coli, such as isopropyl thiogalactopyranoside (IPTG), have been frequently used to trigger the protein expression during early exponential growth phase. This practice has many drawbacks, including high cost and toxicity of IPTG, complex operating procedure, and non-uniform protein expression pattern (some cells in the population do not express recombinant proteins). A few auto-inducible protein expression systems have been developed recently to overcome some of these limitations, but they required use of an additional plasmid or presence of large (a few kilobases) DNA part to be functional, making plasmid construction to be difficult, especially when multiple genes need to be expressed. In this study, by using RNA sequencing, we identified a short, endogenous promoter (PthrC) that can be auto-induced during early exponential growth phase, and improved its performance by use of native and mutated regulatory elements. We found that the developed mutants of PthrC drove uniform protein expression-close to 100% of cells were fluorescent when green fluorescence protein was used as target protein-and cells carrying them could achieve much higher cell density than those with T7 promoter (PT7), a commonly used inducible promoter. In terms of promoter strength (product protein quantity per cell), the developed promoter mutants can cover a range of strength, from 30 to 150% of maximal strength of PT7. One strong mutant (PthrC3_8) was found to work well at a large range of temperature (22, 30, 37 °C) and in various media, and was also confirmed to cause less stress to host cell than PT7 when they were used to express a toxic protein. We foresee that PthrC3 and its mutants will be useful genetic parts for various applications including metabolic engineering and biocatalysis.
journal_name
Appl Microbiol Biotechnoljournal_title
Applied microbiology and biotechnologyauthors
Anilionyte O,Liang H,Ma X,Yang L,Zhou Kdoi
10.1007/s00253-018-9141-zsubject
Has Abstractpub_date
2018-08-01 00:00:00pages
7007-7015issue
16eissn
0175-7598issn
1432-0614pii
10.1007/s00253-018-9141-zjournal_volume
102pub_type
杂志文章abstract::The helix-loop-helix (HLH) family of transcriptional factors is a key player in a wide range of developmental processes in organisms from mammals to microbes. We previously identified the bHLH transcription factor SclR in Aspergillus oryzae and found that the loss of SclR function led to significant phenotypic changes...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-017-8588-7
更新日期:2018-01-01 00:00:00
abstract::Copper (Cu(II)) and nickel (Ni(II)) are often encountered in wastewaters. This study investigated the individual toxic effects of long-term addition of Cu(II) and Ni(II) on the biochemical properties of aerobic granules in sequencing batch reactors (SBRs). The biochemical properties of aerobic granules were characteri...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-010-2467-9
更新日期:2010-05-01 00:00:00
abstract::Phytoplankton blooms are predictable features of marine and freshwater habitats. Despite a good knowledge base of the environmental factors controlling blooms, complex interactions between the bacterial and archaeal communities and phytoplankton bloom taxa are only now emerging. Here, the current research on bacterial...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-017-8330-5
更新日期:2017-06-01 00:00:00
abstract::Twenty-three genotypes of maitake (Grifola frondosa) from Asia, North America and Europe were compared for biological efficiency (BE), quality and crop cycle time. Significant differences among lines were found for BE and crop cycle time when mushrooms were produced on nutrient-supplemented (15% millet, 10% wheat bran...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-001-0875-6
更新日期:2002-02-01 00:00:00
abstract::Microbial natural products, in particular, the ones produced by the members of the order Actinomycetales, will continue to represent an important route to the discovery of novel classes of bioactive compounds. As a result, the search for and discovery of lesser-known and/or novel actinomycetes is of significant intere...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-012-3898-2
更新日期:2012-03-01 00:00:00
abstract::The aim of the present study was to develop a new cell modification method to facilitate the cell separation from broth. In order to reduce the transfer limitation of substrate and product caused by general immobilization methods in the following biotransformation of glycerol, the carboxyl-functioned superparamagnetic...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-013-4723-2
更新日期:2013-05-01 00:00:00
abstract::Carboligations catalyzed by aldolases or thiamine diphosphate (ThDP)-dependent enzymes are well-known in biocatalysis to deliver enantioselective chain elongation reactions. A pyruvate-dependent aldolase (2-oxo-3-deoxy-6-phosphogluconate aldolase [EDA]) introduces a chiral center when reacting with the electrophile, g...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-018-9259-z
更新日期:2018-10-01 00:00:00
abstract::The polyene macrolide rimocidin, produced by Streptomyces rimosus M527, was found to be highly effective against a broad range of fungal plant pathogens. Current understanding of the regulatory mechanism of rimocidin biosynthesis and morphological differentiation in S. rimosus M527 is limited. NsdA is considered a neg...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-020-10955-8
更新日期:2020-12-01 00:00:00
abstract::Streptococcus mutans is the main cause of dental caries. We evaluated the therapeutic effect of variable regions of a llama heavy chain antibody fragments directed against S. mutans named S36-VHH (S for Streptococcus) alone or fused with glucose oxidase (GOx) from Aspergillus niger. Western blot analysis and ELISA rev...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-006-0347-0
更新日期:2006-10-01 00:00:00
abstract::2-Hydroxyalkanoates (2HAs) have become the new monomeric constituents of bacterial polyhydroxyalkanoates (PHAs). PHAs containing 2HA monomers, lactate (LA), glycolate (GL), and 2-hydroxybutyrate (2HB) can be synthesized by engineered microbes in which the broad substrate specificities of PHA synthase and propionyl-CoA...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-013-5120-6
更新日期:2013-09-01 00:00:00
abstract::In natural settings, anaerobic digestion can take place in a wide temperature range, but industrial digesters are usually operated under either mesophilic (~35 °C) or thermophilic (~55 °C) conditions. The ability of anaerobic digestion microbiota to switch from one operating temperature to the other remains poorly doc...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-015-7046-7
更新日期:2016-02-01 00:00:00
abstract::The capability of lactic acid bacteria (LAB) to produce exopoly- and oligosaccharides was and is the subject of expanding research efforts. Due to their physicochemical properties and health-promoting potential, exopoly- and oligosaccharides from food-grade LAB can be used in the food and other industries and may have...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-006-0469-4
更新日期:2006-08-01 00:00:00
abstract::The utilization of monochlorobenzoate isomers (2-, 3- and 4-chlorobenzoate) by anaerobic microbial consortia in River Nile sediments was systematically evaluated under denitrifying, Fe-reducing, sulfidogenic and methanogenic conditions. Loss of all three chlorobenzoates was noted in denitrifying cultures; furthermore,...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/BF02431930
更新日期:1995-10-01 00:00:00
abstract::Homologous recombination-based recombineering is a widely used DNA cloning and modification technique; recombineering efficiency improvement would be helpful for high-throughput DNA manipulation. Escherichia coli primase DnaG variants, such as DnaG Q576A and DnaG K580A, increase the recombineering efficiency via impai...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-019-09744-9
更新日期:2019-04-01 00:00:00
abstract::Polygalacturonases (PG) have evolved in the past years from a pectinase "simply" being used for food processing to an important parameter in plant-fungal interaction. PG-inhibiting proteins (PGIP) that are synthesised in plants as a specific response to PGs of pathogenic fungi, have become a focus as a possible target...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s002530051628
更新日期:2000-04-01 00:00:00
abstract::Antibiotic resistance is a global challenge and represents a growing threat on human health worldwide. Wastewater treatment plants (WWTPs) are generally considered as hotspots for control and/or dissemination of antibiotic resistance. The role of antibiotics, antibacterial residues, and heavy metals played on the evol...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-015-6404-9
更新日期:2015-05-01 00:00:00
abstract::Some strains belonging to the genera Citrobacter and Enterobacter have been reported to produce chitin/chitosan-like bioflocculants (BFs) from acetate. In this study, to investigate the distribution of the BF-producing potential in the genus Citrobacter and to screen stably and highly BF-producing strains, we obtained...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-012-4668-x
更新日期:2013-11-01 00:00:00
abstract::The human antimicrobial peptide LL-37 is a cationic peptide with antimicrobial activity against both Gram-positive and Gram-negative microorganisms. This work describes the development of an expression system based on Escherichia coli capable of high production of the recombinant LL-37. The fusion protein Trx-LL-37 wa...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-010-2736-7
更新日期:2010-09-01 00:00:00
abstract::Cell-surface functional groups (amino, carboxyl, hydroxyl, as well as phosphate) were chemically modified in various ways to enhance the demulsification capability of the demulsifying bacteria Alcaligenes sp. S-XJ-1. Results demonstrated that the demulsifying activity was significantly inhibited by amino enrichment wi...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-017-8111-1
更新日期:2017-05-01 00:00:00
abstract::A Lactobacillus brevis strain with the ability to synthesize butanol from glucose was constructed by metabolic engineering. The genes crt, bcd, etfB, etfA, and hbd, composing the bcs-operon, and the thl gene encode the enzymes of the lower part of the clostridial butanol pathway (crotonase, butyryl-CoA-dehydrogenase, ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-010-2480-z
更新日期:2010-06-01 00:00:00
abstract::Recent work has shed light on the abundance and diversity of D-amino acids in bacterial extracellular/periplasmic molecules, bacterial cell culture, and bacteria-rich environments. Within the extracellular/periplasmic space, D-amino acids are necessary components of peptidoglycan, and disruption of their synthesis lea...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-014-5726-3
更新日期:2014-06-01 00:00:00
abstract::Polyhydroxyalkanoate (PHA) synthases catalyze chain transfer (CT) reaction after polymerization reaction of PHA by transferring PHA chain from PHA synthase to a CT agent, resulting in covalent bonding of CT agent to PHA chain at the carboxyl end. Previous studies have shown that poly(ethylene glycol) (PEG) is an effec...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-010-2601-8
更新日期:2010-07-01 00:00:00
abstract::Clostridium tyrobutyricum is a promising organism for butyrate and n-butanol production, but cannot grow on sucrose. Three genes (scrA, scrB, and scrK) involved in the sucrose catabolic pathway, along with an aldehyde/alcohol dehydrogenase gene, were cloned from Clostridium acetobutylicum and introduced into C. tyrobu...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-017-8200-1
更新日期:2017-05-01 00:00:00
abstract::Microsomal fraction of fungal cells grabs the attention of many researchers for it contains enzymes that play a role in biotechnologically relevant processes. Microsomal enzymes, namely, CYP450s, were shown to metabolize a wide range of xenobiotic compounds, including PAHs, PCBs, dioxins, and endocrine disruptors, and...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-013-5347-2
更新日期:2013-12-01 00:00:00
abstract::At present, more than 100 strains of Lentinula edodes are cultivated on a commercial scale in China. A simple, reliable, and effective method to distinguish some commercial strains of the superior type from other commercial strains is very important for the Lentinula industry. In this study, 23 commercial strains of L...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-008-1671-3
更新日期:2008-11-01 00:00:00
abstract::Batch experiments were conducted to assess the biotransformation potential of four hydrocarbon monoterpenes (d-limonene, alpha-pinene, gamma-terpinene, and terpinolene) and four alcohols (arbanol, linalool, plinol, and alpha-terpineol) under aerobic conditions at 23 degrees C. Both forest-soil extract and enriched cul...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530050770
更新日期:1996-07-01 00:00:00
abstract::During sporulation, Bacillus thuringiensis produces insecticidal crystal inclusions (Cry proteins) encoded by cry genes. In fed-batch cultures (FBCs), spores and Cry protein yields are usually low, so we therefore studied the pattern of metabolic changes occurring in batch cultures and FBCs of a B. thuringiensis strai...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-004-1724-1
更新日期:2005-04-01 00:00:00
abstract::Autohydrogenotrophic batch growth of Ralstonia eutropha H16 was studied in a stirred-tank reactor with nitrate and nitrite as terminal electron acceptors and the sole limiting substrates. Assuming product inhibition by nitrite, saturation kinetics with the two limiting substrates and a simple switching function, which...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-007-0983-z
更新日期:2007-08-01 00:00:00
abstract::Furfural is one of the major inhibitors generated during sugar production from cellulosic materials and, as an aldehyde, inhibits various cellular activities of microorganisms used, leading to prolonged lag time during ethanologenic fermentation. Since Saccharomyces cerevisiae strains tolerant to furfural are of great...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-012-4022-3
更新日期:2012-07-01 00:00:00
abstract::A recombinant form of the sweet-tasting protein thaumatin has been produced in the filamentous fungus Aspergillus niger var. awamori. Expression cassettes containing a synthetic gene encoding thaumatin II were prepared and used to transform Aspergillus niger var. awamori strain NRRL312. Several fungal strains capable ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530051188
更新日期:1998-04-01 00:00:00