Optimization of protease secretion in Bacillus subtilis and Bacillus licheniformis by screening of homologous and heterologous signal peptides.

Abstract:

:Bacillus subtilis and Bacillus licheniformis are widely used for the large-scale industrial production of proteins. These strains can efficiently secrete proteins into the culture medium using the general secretion (Sec) pathway. A characteristic feature of all secreted proteins is their N-terminal signal peptides, which are recognized by the secretion machinery. Here, we have studied the production of an industrially important secreted protease, namely, subtilisin BPN' from Bacillus amyloliquefaciens. One hundred seventy-three signal peptides originating from B. subtilis and 220 signal peptides from the B. licheniformis type strain were fused to this secretion target and expressed in B. subtilis, and the resulting library was analyzed by high-throughput screening for extracellular proteolytic activity. We have identified a number of signal peptides originating from both organisms which produced significantly increased yield of the secreted protease. Interestingly, we observed that levels of extracellular protease were improved not only in B. subtilis, which was used as the screening host, but also in two different B. licheniformis strains. To date, it is impossible to predict which signal peptide will result in better secretion and thus an improved yield of a given extracellular target protein. Our data show that screening a library consisting of homologous and heterologous signal peptides fused to a target protein can identify more-effective signal peptides, resulting in improved protein export not only in the original screening host but also in different production strains.

journal_name

Appl Environ Microbiol

authors

Degering C,Eggert T,Puls M,Bongaerts J,Evers S,Maurer KH,Jaeger KE

doi

10.1128/AEM.01146-10

subject

Has Abstract

pub_date

2010-10-01 00:00:00

pages

6370-6

issue

19

eissn

0099-2240

issn

1098-5336

pii

AEM.01146-10

journal_volume

76

pub_type

杂志文章
  • Fundamental procedures for determining ergosterol content of decaying plant material by liquid chromatography.

    abstract::Portions of published procedures for measurement of ergosterol content of decomposing plants were examined for their influence upon ergosterol yield. Common methods of treatment of plant samples prior to sterol extraction (e.g., oven drying, freezing, lyophilization) led to reduced recoveries of ergosterol (ca. 20 to ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.54.7.1876-1879.1988

    authors: Newell SY,Arsuffi TL,Fallon RD

    更新日期:1988-07-01 00:00:00

  • Thermal resistance of naturally occurring airborne bacterial spores.

    abstract::Simulation of a heat process used in the terminal dry-heat decontamination of the Viking spacecraft is reported. Naturally occurring airborne bacterial spores were collected on Teflon ribbons in selected spacecraft assembly areas and subsequently subjected to dry heat. Thermal inactivation experiments were conducted a...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.36.3.473-479.1978

    authors: Puleo JR,Bergstrom SL,Peeler JT,Oxborrow GS

    更新日期:1978-09-01 00:00:00

  • Diel nitrogen fixation by cyanobacterial surface blooms in sanctuary lake, pennsylvania.

    abstract::Diel nitrogen fixation studies were conducted with assemblages of cyanobacteria sampled from surface blooms on Sanctuary Lake, Pa. The studies were conducted between July and September of 1982 to 1985 by using the acetylene reduction technique. Assemblages with the lowest cell concentrations (0.9 x 10 to 1.0 x 10 cell...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.56.2.466-471.1990

    authors: Storch TA,Saunders GW,Ostrofsky ML

    更新日期:1990-02-01 00:00:00

  • Genotype distribution of human sapoviruses in wastewater in Japan.

    abstract::Human sapovirus sequences were identified in 12 (100%) influent and 7 (58%) effluent wastewater samples collected once a month for a year. The strains were characterized based on their partial capsid gene sequences and classified into 10 genotypes, demonstrating that genetically diverse sapovirus strains infect humans...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.00088-11

    authors: Kitajima M,Haramoto E,Phanuwan C,Katayama H

    更新日期:2011-06-01 00:00:00

  • Nutritional and biochemical characterization of Methanospirillum hungatii.

    abstract::To ascertain its physiological similarity to other methanogenic bacteria, Methanospirillum hungatii, the type species of the genus, was characterized nutritionally and biochemically. Good growth occurred in a medium consisting of mineral salts, cysteine sulfide reducing buffer, and an H2-CO2 (80:20) atmosphere. Additi...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.34.4.371-376.1977

    authors: Ferry JG,Wolfe RS

    更新日期:1977-10-01 00:00:00

  • Development of a solid medium for growth and isolation of axenic microcystis strains (cyanobacteria).

    abstract::Solid media on a base of B-12 or CB medium with agarose or agarose of low melting temperature were developed for the cultivation of Microcystis species. The media with 0.4% gel showed the highest number of CFU, and increasing the gel concentration resulted in a reduction of the number of CFU. There was no difference i...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.55.10.2569-2571.1989

    authors: Shirai M,Matumaru K,Ohotake A,Takamura Y,Aida T,Nakano M

    更新日期:1989-10-01 00:00:00

  • Quantitative detection of perchlorate-reducing bacteria by real-time PCR targeting the perchlorate reductase gene.

    abstract::A quantitative real-time PCR assay targeting the pcrA gene, encoding the catalytic subunit of perchlorate reductase, detected pcrA genes from perchlorate-reducing bacteria in three different genera and from soil microbial communities. Partial pcrA sequences indicated differences in the composition of perchlorate-reduc...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01658-07

    authors: Nozawa-Inoue M,Jien M,Hamilton NS,Stewart V,Scow KM,Hristova KR

    更新日期:2008-03-01 00:00:00

  • Interaction of Aeromonas strains with lactic acid bacteria via Caco-2 cells.

    abstract::The genus Aeromonas includes some species that have now been identified as human pathogens of significant medical importance. We investigated the ability of 13 selected Aeromonas strains belonging to nine species isolated from clinical cases (n = 5), environmental waters (n = 5), and fish (n = 3) to adhere to and tran...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.03200-13

    authors: Hatje E,Neuman C,Katouli M

    更新日期:2014-01-01 00:00:00

  • Serological diversity within a terrestrial ammonia-oxidizing population.

    abstract::Fluorescent antibodies (FAs) prepared against 16 ammonia-oxidizing nitrifying bacteria were examined as to cross-reactivity in heterologous FA staining tests. Virtually all cross-reactions to the seven Nitrosomonas FAs were confined to the Nitrosomonas isolates. The five Nitrosospira, three Nitrosolobus, and one Nitro...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.36.4.589-593.1978

    authors: Belser LW,Schmidt EL

    更新日期:1978-10-01 00:00:00

  • Transformation of Escherichia coli with DNA from Saccharomyces cerevisiae cell lysates.

    abstract::We developed a system to monitor the transfer of heterologous DNA from a genetically manipulated strain of Saccharomyces cerevisiae to Escherichia coli. This system is based on a yeast strain that carries multiple integrated copies of a pUC-derived plasmid. The bacterial sequences are maintained in the yeast genome by...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.65.12.5303-5306.1999

    authors: Adam AC,González-Blasco G,Rubio-Texeira M,Polaina J

    更新日期:1999-12-01 00:00:00

  • Response of Vibrio cholerae to Low-Temperature Shifts: CspV Regulation of Type VI Secretion, Biofilm Formation, and Association with Zooplankton.

    abstract:UNLABELLED:The ability to sense and adapt to temperature fluctuation is critical to the aquatic survival, transmission, and infectivity of Vibrio cholerae, the causative agent of the disease cholera. Little information is available on the physiological changes that occur when V. cholerae experiences temperature shifts....

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.00807-16

    authors: Townsley L,Sison Mangus MP,Mehic S,Yildiz FH

    更新日期:2016-06-30 00:00:00

  • Isolation and Partial Characterization of Two Aeromonas hydrophila Bacteriophages.

    abstract::Two Aeromonas hydrophila bacteriophages, Aeh1 and Aeh2, were isolated from sewage. Both phages showed binal symmetry. The dimensions of A. hydrophila phages Aeh1 and Aeh2 differed from those of the other Aeromonas phages. Also, phage Aeh2 was the largest Aeromonas phage studied to date. Phage Aeh1 formed small, clear ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.45.5.1670-1676.1983

    authors: Chow MS,Rouf MA

    更新日期:1983-05-01 00:00:00

  • Binding of virus-like particles of Norwalk virus to romaine lettuce veins.

    abstract::Noroviruses (NoV) annually cause millions of cases of gastrointestinal disease in the United States. NoV are associated with raw shellfish outbreaks, particularly oysters, which are thought to bioaccumulate NoV particles during the filter-feeding process. NoV outbreaks, however, have also been known to occur from othe...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01566-10

    authors: Gandhi KM,Mandrell RE,Tian P

    更新日期:2010-12-01 00:00:00

  • Sulfate Ester Detergent Degradation in Pseudomonas aeruginosa Is Subject to both Positive and Negative Regulation.

    abstract::Bacteria using toxic chemicals, such as detergents, as growth substrates face the challenge of exposing themselves to cell-damaging effects that require protection mechanisms, which demand energy delivered from catabolism of the toxic compound. Thus, adaptations are necessary for ensuring the rapid onset of substrate ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01352-19

    authors: Panasia G,Oetermann S,Steinbüchel A,Philipp B

    更新日期:2019-11-14 00:00:00

  • Kinetic explanation for accumulation of nitrite, nitric oxide, and nitrous oxide during bacterial denitrification.

    abstract::The kinetics of denitrification and the causes of nitrite and nitrous oxide accumulation were examined in resting cell suspensions of three denitrifiers. An Alcaligenes species and a Pseudomonas fluorescens isolate characteristically accumulated nitrite when reducing nitrate; a Flavobacterium isolate did not. Nitrate ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.42.6.1074-1084.1981

    authors: Betlach MR,Tiedje JM

    更新日期:1981-12-01 00:00:00

  • Spectrum of the reductive dehalogenation activity of desulfitobacterium frappieri PCP-1

    abstract::Desulfitobacterium frappieri PCP-1 was induced for ortho- and para-dechlorinating activities by different chlorophenols. Dehalogenation rates ranging from 25 to 1,158 nmol/min/mg of cell protein were observed according to the chlorophenol tested and the position of the chlorine removed. D. frappieri shows a broad subs...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.64.11.4603-4606.1998

    authors: Dennie D,Gladu I I,Lepine F,Villemur R,Bisaillon J,Beaudet R

    更新日期:1998-11-01 00:00:00

  • Habitat-specific diversity of Borrelia burgdorferi sensu lato in Europe, exemplified by data from Latvia.

    abstract::The distribution of Borrelia burgdorferi sensu lato genospecies in questing Ixodes ricinus ticks from ecologically distinct habitats in Latvia was analyzed. A significant variation in the frequency of the genospecies across sites was observed, pointing to the importance of the host community in the ecology of Lyme bor...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/aem.69.5.3008-3010.2003

    authors: Etti S,Hails R,Schäfer SM,De Michelis S,Sewell HS,Bormane A,Donaghy M,Kurtenbach K

    更新日期:2003-05-01 00:00:00

  • Effect of culture conditions on ergosterol as an indicator of biomass in the aquatic hyphomycetes.

    abstract::Ergosterol is a membrane component specific to fungi that can be used to estimate fungal biomass using appropriate factors of conversion. Our objectives were to determine the limits of use of ergosterol content as a measure of biomass for aquatic hyphomycetes, and to evaluate a previously established ergosterol-to-bio...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.67.5.2051-2055.2001

    authors: Charcosset JY,Chauvet E

    更新日期:2001-05-01 00:00:00

  • Crystal Structure of TetR Family Repressor AlkX from Dietzia sp. Strain DQ12-45-1b Implicated in Biodegradation of n-Alkanes.

    abstract::n-Alkanes are ubiquitous in nature and are widely used by microorganisms as carbon sources. Alkane hydroxylation by alkane monooxygenases is a critical step in the aerobic biodegradation of n-alkanes, which plays important roles in natural alkane attenuation and is used in industrial and environmental applications. Th...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01447-17

    authors: Liang JL,Gao Y,He Z,Nie Y,Wang M,JiangYang JH,Zhang XC,Shu WS,Wu XL

    更新日期:2017-10-17 00:00:00

  • Variation in Sensitivity of Gaeumannomyces graminis to Antibiotics Produced by Fluorescent Pseudomonas spp. and Effect on Biological Control of Take-All of Wheat.

    abstract::Isolates of Gaeumannomyces graminis var. tritici, the causal agent of take-all of wheat, varied in sensitivity in vitro to the antibiotics phenazine-1-carboxylic acid (PCA) and 2,4-diacetylphloroglucinol (Phl) produced by fluorescent Pseudomonas spp. shown previously to have potential for biological control of this pa...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.61.7.2554-2559.1995

    authors: Mazzola M,Fujimoto DK,Thomashow LS,Cook RJ

    更新日期:1995-07-01 00:00:00

  • New method for isolating barophiles from intestinal contents of deep-sea fishes retrieved from the abyssal zone.

    abstract::We devised a new method (the dorayaki method) using marine agar under in situ pressures to isolate barophilic bacteria from the intestinal contents of three deep-sea fishes (two Coryphaenoides yaquinae samples and one Ilyophis sp. sample) retrieved from depths of 4,700 to 6,100 m in the Northwest Pacific Ocean. All 10...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.60.11.4210-4212.1994

    authors: Nakayama A,Yano Y,Yoshida K

    更新日期:1994-11-01 00:00:00

  • Long-Term Fungal Inhibition by Pisum sativum Flour Hydrolysate during Storage of Wheat Flour Bread.

    abstract::In order to identify antifungal compounds from natural sources to be used as ingredients in the bakery industry, water/salt-soluble extracts (WSE) from different legume flour hydrolysates obtained by the use of a fungal protease were assayed against Penicillium roqueforti DPPMAF1. The agar diffusion assays allowed the...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.04088-14

    authors: Rizzello CG,Lavecchia A,Gramaglia V,Gobbetti M

    更新日期:2015-06-15 00:00:00

  • Genomic Variability of the Xanthomonas Pathovar mangiferaeindicae, Agent of Mango Bacterial Black Spot.

    abstract::The genetic diversity of 138 strains of the Xanthomonas pathovar mangiferaeindicae, which were isolated from three different hosts (mango, ambarella, and pepper tree) in 14 different countries, was assessed with restriction fragment length polymorphism markers. An analysis of patterns obtained by hybridization with an...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.63.1.246-253.1997

    authors: Gagnevin L,Leach JE,Pruvost O

    更新日期:1997-01-01 00:00:00

  • Diversity of basidiomycetes in michigan agricultural soils.

    abstract::We analyzed the communities of soil basidiomycetes in agroecosystems that differ in tillage history at the Kellogg Biological Station Long-Term Ecological Research site near Battle Creek, Michigan. The approach combined soil DNA extraction through a bead-beating method modified to increase recovery of fungal DNA, PCR ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.00826-06

    authors: Lynch MD,Thorn RG

    更新日期:2006-11-01 00:00:00

  • Cloning and characterization of the lactococcal plasmid-encoded type II restriction/modification system, LlaDII.

    abstract::The LlaDII restriction/modification (R/M) system was found on the naturally occurring 8.9-kb plasmid pHW393 in Lactococcus lactis subsp. cremoris W39. A 2.4-kb PstI-EcoRI fragment inserted into the Escherichia coli-L. lactis shuttle vector pCI3340 conferred to L. lactis LM2301 and L. lactis SMQ86 resistance against re...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.64.7.2424-2431.1998

    authors: Madsen A,Josephsen J

    更新日期:1998-07-01 00:00:00

  • Sucrose synthesis in the nitrogen-fixing Cyanobacterium Anabaena sp. strain PCC 7120 is controlled by the two-component response regulator OrrA.

    abstract::The filamentous, nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120 accumulates sucrose as a compatible solute against salt stress. Sucrose-phosphate synthase activity, which is responsible for the sucrose synthesis, is increased by salt stress, but the mechanism underlying the regulation of sucrose synthesis...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01501-14

    authors: Ehira S,Kimura S,Miyazaki S,Ohmori M

    更新日期:2014-09-01 00:00:00

  • Clostridium acetobutylicum Protoplast Formation and Regeneration.

    abstract::Techniques and media for the production and regeneration of stable Clostridium acetobutylicum protoplasts are described. ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.43.3.719-721.1982

    authors: Allcock ER,Reid SJ,Jones DT,Woods DR

    更新日期:1982-03-01 00:00:00

  • Odorous Sulfur Compounds Emitted during Production of Compost Used as a Substrate in Mushroom Cultivation.

    abstract::Large-scale composting facilities are known to cause environmental problems, mainly through pungent air emitted by composting material. In air samples taken above stacks set up to prepare compost used as a substrate in mushroom cultivation, several volatile compounds were identified by means of the coupled techniques ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.56.1.176-180.1990

    authors: Derikx PJ,Op Den Camp HJ,van der Drift C,van Griensven LJ,Vogels GD

    更新日期:1990-01-01 00:00:00

  • Genetic characterization and physiological role of endopeptidase O from Lactobacillus helveticus CNRZ32.

    abstract::A previously identified insert expressing an endopeptidase from a Lactobacillus helveticus CNRZ32 genomic library was characterized. Nucleotide sequence analysis revealed an open reading frame of 1,941 bp encoding a putative protein of 71.2 kDa which contained a zinc-protease motif. Protein homology searches revealed ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.64.9.3411-3415.1998

    authors: Chen YS,Steele JL

    更新日期:1998-09-01 00:00:00

  • Biofilm formation on reverse osmosis membranes is initiated and dominated by Sphingomonas spp.

    abstract::The initial formation and spatiotemporal development of microbial biofilm layers on surfaces of new and clean reverse osmosis (RO) membranes and feed-side spacers were monitored in situ using flow cells placed in parallel with the RO system of a full-scale water treatment plant. The feed water of the RO system had bee...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01998-09

    authors: Bereschenko LA,Stams AJ,Euverink GJ,van Loosdrecht MC

    更新日期:2010-04-01 00:00:00