Abstract:
:Due to their high secretion capacity, Gram-positive bacteria from the genus Bacillus are important expression hosts for the high-yield production of enzymes in industrial biotechnology; however, to date, strains from only few Bacillus species are used for enzyme production at industrial scale. Herein, we introduce Paenibacillus polymyxa DSM 292, a member of a different genus, as a novel host for secretory protein production. The model gene cel8A from Clostridium thermocellum was chosen as an easily detectable reporter gene with industrial relevance to demonstrate heterologous expression and secretion in P. polymyxa. The yield of the secreted cellulase Cel8A protein was increased by optimizing the expression medium and testing several promoter sequences in the expression plasmid pBACOV. Quantitative mass spectrometry was used to analyze the secretome in order to identify promising new promoter sequences from the P. polymyxa genome itself. The most abundantly secreted host proteins were identified, and the promoters regulating the expression of their corresponding genes were selected. Eleven promoter sequences were cloned and tested, including well-characterized promoters from Bacillus subtilis and Bacillus megaterium. The best result was achieved with the promoter for the hypothetical protein PPOLYM_03468 from P. polymyxa. In combination with the optimized expression medium, this promoter enabled the production of 5475 U/l of Cel8A, which represents a 6.2-fold increase compared to the reference promoter PaprE. The set of promoters described in this work covers a broad range of promoter strengths useful for heterologous expression in the new host P. polymyxa.
journal_name
Appl Microbiol Biotechnoljournal_title
Applied microbiology and biotechnologyauthors
Heinze S,Zimmermann K,Ludwig C,Heinzlmeir S,Schwarz WH,Zverlov VV,Liebl W,Kornberger Pdoi
10.1007/s00253-018-9369-7subject
Has Abstractpub_date
2018-12-01 00:00:00pages
10147-10159issue
23eissn
0175-7598issn
1432-0614pii
10.1007/s00253-018-9369-7journal_volume
102pub_type
杂志文章abstract::The accumulation of toxic inorganic nitrogen is one of the major water quality problems in intensive aquaculture systems, thus the N removal in aquaculture systems is an important issue for the sustainable development of aquaculture. To understand one of the major microbial N removal processes, anaerobic ammonium oxid...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-016-7690-6
更新日期:2016-10-01 00:00:00
abstract::Secretion of fusion proteins composed of cytoplasmic protein dihydrofolate reductase (DHFR) and the Escherichia coli alpha-haemolysin (HlyA) C-terminal sequence was examined through the haemolysin secretion machinery of E. coli. DHFR of various lengths was combined with the HlyA C-terminal region, and both secretion a...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/BF00174844
更新日期:1992-09-01 00:00:00
abstract::Cultivation of Aspergillus terreus ATCC 20542 in a stirred tank bioreactor was performed to induce the biosynthesis of secondary metabolites and provide the bioprocess-related insights into the metabolic capabilities of the investigated strain. The activation of biosynthetic routes was attempted by the diversification...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-015-7157-1
更新日期:2016-04-01 00:00:00
abstract::Fossil fuel reserves depletion, global warming, unrelenting population growth, and costly and problematic waste recycling call for renewable resources of energy and consumer products. As an alternative to the 100 % oil economy, production processes based on biomass can be developed. Huge amounts of lignocellulosic was...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-013-5014-7
更新日期:2013-07-01 00:00:00
abstract::The polyene macrolide rimocidin, produced by Streptomyces rimosus M527, is highly effective against a broad range of fungal plant pathogens, but at low yields. Elicitation is an effective method of stimulating the yield of bioactive secondary metabolites. In this study, the biomass and filtrate of a culture broth of E...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-020-10565-4
更新日期:2020-05-01 00:00:00
abstract::In this study, we employed TiO₂ enrichment and high accuracy liquid chromatography-mass spectrometry-mass spectrometry to identify the phosphoproteome of Clostridium acetobutyicum ATCC824 in acidogenesis and solventogenesis. As many as 82 phosphopeptides in 61 proteins, with 107 phosphorylated sites on serine, threoni...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-012-4156-3
更新日期:2012-07-01 00:00:00
abstract::A comparative study was carried out in anaerobic batch cultures on 20 g/l of either glycerol or glucose using two propionibacteria strains, Propionibacterium acidipropionici and Propionibacterium freudenreichii ssp. shermanii. In all cases, fermentation end-products were the same and consisted of propionic acid as the...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530051638
更新日期:2000-04-01 00:00:00
abstract::His-Asn-His (HNH) proteins are a very common family of small nucleic acid-binding proteins that are generally associated with endonuclease activity and are found in all kingdoms of life. Although HNH endonucleases from mesophiles have been widely investigated, the biochemical functions of HNH endonucleases from thermo...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-016-7568-7
更新日期:2016-09-01 00:00:00
abstract::Pyranose 2-oxidase (P2O) was purified 43-fold to apparent homogeneity from the basidiomycete Phanerochaete chrysosporium using liquid chromatography on phenyl Sepharose, Mono Q (twice) and phenyl Superose. The native enzyme has a molecular mass of about 250 kDa (based on native PAGE) and is composed of four identical ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530050964
更新日期:1997-05-01 00:00:00
abstract::The ligninolytic enzymes of white-rot fungi have a broad substrate specificity and have been implicated in the transformation and mineralization of organopollutants with structural similarities to lignin. This review presents evidence for the involvement of these enzymes in white-rot fungal degradation of munitions wa...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s002530100745
更新日期:2001-10-01 00:00:00
abstract::The bean bug, Riptortus pedestris, is a major agricultural pest that reduces crop quality and value. Chemical pesticides have contributed to pest management, but resistance to these chemicals has significantly limited their use. Alternative strategies with different modes of action, such as entomopathogenic fungi, are...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-016-7734-y
更新日期:2016-10-01 00:00:00
abstract::Removal of tetrachloroethylene (perchloroethylene; C2Cl4) by microbial consortia from two sites with different C2Cl4 exposure histories was examined in a bench-scale anaerobic column bioreactor. It was hypothesized that optimal removal would be observed in the reactor packed with sediments having an extensive exposure...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530051074
更新日期:1997-09-01 00:00:00
abstract::We report an immuno-magnetic separation system developed by the immobilization of pathogen-specific antibodies on the core-shell magnetic beads. The magnetic beads were grafted with glycidylmethacrylate (GMA) using surface-initiated atom transfer radical polymerization (SI-ATRP). For immuno-magnetic separation (IMS) o...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-013-5231-0
更新日期:2013-11-01 00:00:00
abstract::Previous investigations have reported that bacterial suspension cultures grow to higher stationary concentrations in space flight than on Earth; however, none of these investigations included extensive ground controls under varied inertial conditions. This study includes extensive controls and cell-growth data taken a...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530051386
更新日期:1999-02-01 00:00:00
abstract::The medicinal mushroom Agaricus blazei produced high amounts of laccase (up to 5,000 units l(-1)) in a complex, agitated liquid medium based on tomato juice, while only traces of the enzyme (<100 units l(-1)) were detected in synthetic glucose-based medium. Purification of the enzyme required three chromatographic ste...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-004-1861-6
更新日期:2005-05-01 00:00:00
abstract::Streptolydigin, a secondary metabolite produced by Streptomyces lydicus, is a potent inhibitor of bacterial RNA polymerases. It has been suggested that streptolydigin biosynthesis is associated with polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS). Thus, there is great interest in understanding the...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-009-1872-4
更新日期:2009-05-01 00:00:00
abstract::The 5'-untranslated leader sequence (UTLS) of the slpA gene from Lactobacillus acidophilus contributes to mRNA stabilization by producing a 5' stem and loop structure, and a high-level expression system for the lactic acid bacteria was developed using the UTLS in this study. A plasmid, which expresses alpha-amylase un...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-006-0477-4
更新日期:2006-11-01 00:00:00
abstract::Microbial cell factories offer an economic approach for synthesizing "natural'" aromatic flavor compounds. During their fermentation process, the inefficient synthesis pathway and product cytotoxicity are the major barriers to the high-level production. This study combined metabolic engineering and tolerance engineeri...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-020-10991-4
更新日期:2020-12-01 00:00:00
abstract::Sulfadiazine (SDZ) is an antibiotic frequently administered to livestock, and it alters microbial communities when entering soils with animal manure, but understanding the interactions of these effects to the prevailing climatic regime has eluded researchers. A climatic factor that strongly controls microbial activity...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-014-5717-4
更新日期:2014-01-01 00:00:00
abstract::A precise and scarless genome excision method, employing the Cre/loxP system in concert with double-strand break (DSB)-stimulated intramolecular recombination was developed. The DSBs were mediated by the restriction endonuclease, I-SceI. It permitted multiple deletions of independent 14-, 43-, and 10-kb-long genomic r...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-005-1976-4
更新日期:2005-11-01 00:00:00
abstract::Relative quantification real-time PCR was used to quantify several bacterial species in ruminal samples from two lactating cows, each sampled 3 h after feeding on two successive days. Abundance of each target taxon was calculated as a fraction of the total 16S rRNA gene copies in the samples, using taxon-specific and ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-006-0802-y
更新日期:2007-05-01 00:00:00
abstract::Enterobacter sp. strain FL was newly isolated from activated sludge and exhibited significant capability of auto-aggregation as well as aerobic denitrification. The removal efficiencies of NO3--N, total nitrogen (TN), and TOC by strain FL in batch culture reached 94.6, 63.9, and 72.5% in 24 h, respectively. The produc...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-017-8720-8
更新日期:2018-02-01 00:00:00
abstract::Laccases are blue multicopper oxidases with potential applications in environmental and industrial biotechnology. In this study, a new bacterial laccase gene of 1.32 kb was obtained from a marine microbial metagenome of the South China Sea by using a sequence screening strategy. The protein (named as Lac15) of 439 ami...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-010-2934-3
更新日期:2011-02-01 00:00:00
abstract::This study represents two different large-scale proteomic experiments analyzing the antibiotic response and the mechanisms of production of β-lactamases in the nosocomial pathogen Stenotrophomonas maltophilia. Two-dimensional gel electrophoresis on the cytoplasmic protein fraction, together with iTRAQ® differential la...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-012-4167-0
更新日期:2012-08-01 00:00:00
abstract::Flavour and fragrance compounds are extremely important for food, feed, cosmetic and pharmaceutical industries. In the last decades, due to the consumer's increased trend towards natural products, a great interest in natural aroma compounds has arisen to the detriment of chemically synthesised ones. Recently, solid st...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-018-9157-4
更新日期:2018-09-01 00:00:00
abstract::In this study, an evolved Saccharomyces cerevisiae HJ7-14 with high ability of algae-based ethanol production was characterized by multi-omic approaches. Genome sequencing of the HJ7-14 revealed a point mutation in the GAL83 gene (G703A) involved in the catabolite repression as well as the galactose metabolism. Cultur...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-018-9306-9
更新日期:2018-10-01 00:00:00
abstract::Sulfur-containing aroma compounds are key contributors to the flavour of a diverse range of foods and beverages, such as wine. The tropical fruit characters of Sauvignon Blanc wines are attributed to the presence of the aromatic thiols 3-mercaptohexan-1-ol (3-MH), its acetate ester 3-mercaptohexyl acetate (3-MHA), and...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-015-6833-5
更新日期:2015-11-01 00:00:00
abstract::Gene therapy, i.e., the expression in cells of genetic material with therapeutic activity, holds great promise for the treatment of human diseases. A delivery vehicle (vector), of either viral or non-viral origin, must be used to carry the foreign gene into a cell. Viral vectors take advantage of the facile integratio...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-003-1321-8
更新日期:2003-07-01 00:00:00
abstract::Indigo is an insoluble blue dye historically used for dyeing textiles. A traditional approach for indigo dyeing involves microbial reduction of polygonum indigo to solubilize it under alkaline conditions; however, the mechanism by which microorganisms reduce indigo remains poorly understood. Here, we aimed to identify...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-018-9284-y
更新日期:2018-11-01 00:00:00
abstract::Extracellular polymeric substance (EPS) is proposed to facilitate calcium ion supersaturation through its nucleation effect during the microbially induced calcium carbonate precipitation (MICP) process. However, the supersaturation effect of Ca2+ via EPS in MICP has not been clearly demonstrated. Enhanced exopolysacch...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-017-8372-8
更新日期:2017-08-01 00:00:00