Engineered cyanophycin synthetase (CphA) from Nostoc ellipsosporum confers enhanced CphA activity and cyanophycin accumulation to Escherichia coli.

Abstract:

:The cyanophycin (CGP) synthetase gene (cphANE1) of the transposon-induced argL mutant NE1 of the cyanobacterium Nostoc ellipsosporum, which exhibits a CGP-leaky phenotype during diazotrophical growth, was cloned and expressed in Escherichia coli strain TOP10. Its amino acid sequence exhibited high similarities to CphAs of other cyanobacteria. Recombinant cells of E. coli, which harbored a fragment comprising the complete cphANE1 gene plus 400 bp of its downstream region in colinear orientation to the lacZ promoter, accumulated CGP up to 17 and 8.5% (wt/wt) of cellular dry matter (CDM) if cultivated in complex medium in the presence or absence of isopropyl-beta-D-thiogalactopyranoside, respectively. Two truncated CphAs, lacking 31 (CphANE1del96) or 59 (CphANE1del180) amino acids of the C-terminal region, were derived from cphANE1 by deleting 96 or 180 bp from its 3' region through the introduction of stop codons. In comparison to the wild-type gene, cphANE1del96 conferred about 2.1- to 2.2-fold-higher enzyme activity (up to 5.75 U/mg protein) on E. coli. Furthermore, these cells accumulated about twofold more CGP (up to 34.5% [wt/wt] of CDM) than cells expressing the wild-type gene. An engineered CphA possessing significantly enhanced activity and conferring the highest CGP content on E. coli is demonstrated. In contrast, CphANE1del180 was inactive and did not confer CGP accumulation on E. coli. Interestingly, a short conserved stretch of 4 to 5 hydrophobic amino acids is located in the protein region present in CphANE1del96 but absent in CphANE1del180. In addition, CphANE1 and CphANE1del96 are, besides CphA from Acinetobacter baylyi, the only CphAs exhibiting rigid substrate specificities that do not enable the incorporation of lysine instead of arginine into CGP.

journal_name

Appl Environ Microbiol

authors

Hai T,Frey KM,Steinbüchel A

doi

10.1128/AEM.01132-06

subject

Has Abstract

pub_date

2006-12-01 00:00:00

pages

7652-60

issue

12

eissn

0099-2240

issn

1098-5336

pii

AEM.01132-06

journal_volume

72

pub_type

杂志文章
  • Probiotic Lactobacillus rhamnosus Reduces Organophosphate Pesticide Absorption and Toxicity to Drosophila melanogaster.

    abstract::Organophosphate pesticides used in agriculture can pose health risks to humans and wildlife. We hypothesized that dietary supplementation with Lactobacillus, a genus of commensal bacteria, would reduce absorption and toxicity of consumed organophosphate pesticides (parathion and chlorpyrifos [CP]). Several Lactobacill...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01510-16

    authors: Trinder M,McDowell TW,Daisley BA,Ali SN,Leong HS,Sumarah MW,Reid G

    更新日期:2016-09-30 00:00:00

  • Tailing of thermal inactivation curve of Aspergillus niger spores.

    abstract::The nonlinear thermal inactivation of Aspergillus niger spores in phosphate-citrate buffer was studied. The thermal inactivation pattern of the spore consisted of a shoulder, an accelerated decline, and a tail at various constant temperatures around 60 degrees C. The pattern fitted a thermotolerant subpopulation model...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.62.10.3745-3749.1996

    authors: Fujikawa H,Itoh T

    更新日期:1996-10-01 00:00:00

  • Improved specimen coating technique for scanning electron microscope observation of decomposer microorganisms.

    abstract::Sputter coating of leaf litter microbe samples provides scanning electron microscope images with greater information content than either vacuum evaporation of thin metal coatings or tissue conductance. ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.31.2.313-315.1976

    authors: Draggan S

    更新日期:1976-02-01 00:00:00

  • Expression of the isoamylase gene of Flavobacterium odoratum KU in Escherichia coli and identification of essential residues of the enzyme by site-directed mutagenesis.

    abstract::The isoamylase gene from Flavobacterium odoratum KU was cloned into and expressed in Escherichia coli JM109. The promoter of the gene was successful in E. coli, and the enzyme produced was excreted into the culture medium, depending on the amount of the enzyme expressed. The enzyme found in the culture medium showed a...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.65.9.4163-4170.1999

    authors: Abe J,Ushijima C,Hizukuri S

    更新日期:1999-09-01 00:00:00

  • Effect of different holding regimens on the intestinal microflora of herring (Clupea harengus) larvae.

    abstract::The aerobic intestinal microflora of 2-week-old herring (Clupea harengus) larvae was characterized by using conventional microbiological methods and electron microscopy. Larvae were hatched and kept in filtered seawater or in seawater with penicillin and streptomycin. The gastrointestinal tract of herring larvae is es...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.58.2.461-470.1992

    authors: Hansen GH,Strøm E,Olafsen JA

    更新日期:1992-02-01 00:00:00

  • Use of 16S-23S rRNA intergenic spacer region PCR and repetitive extragenic palindromic PCR analyses of Escherichia coli isolates to identify nonpoint fecal sources.

    abstract::Despite efforts to minimize fecal input into waterways, this kind of pollution continues to be a problem due to an inability to reliably identify nonpoint sources. Our objective was to find candidate source-specific Escherichia coli fingerprints as potential genotypic markers for raw sewage, horses, dogs, gulls, and c...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/aem.69.8.4942-4950.2003

    authors: Seurinck S,Verstraete W,Siciliano SD

    更新日期:2003-08-01 00:00:00

  • Ammonium Limitation Results in the Loss of Ammonia-Oxidizing Activity in Nitrosomonas europaea.

    abstract::The effects of limiting concentrations of ammonium on the metabolic activity of Nitrosomonas europaea, an obligate ammonia-oxidizing soil bacterium, were investigated. Cells were harvested during late logarithmic growth and were incubated for 24 h in growth medium containing 0, 15, or 50 mM ammonium. The changes in ni...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.64.4.1514-1521.1998

    authors: Stein LY,Arp DJ

    更新日期:1998-04-01 00:00:00

  • Rapid and simple cryopreservation of anaerobic ammonium-oxidizing bacteria.

    abstract::A quick and simple protocol for long-term cryopreservation of anaerobic ammonium-oxidizing bacteria (anammox bacteria) was developed. After 29 weeks of preservation at -80°C, activity recovery for all tested cultures under at least one of the applied sets of preservation conditions was observed. Growth recovery was al...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.07501-11

    authors: Heylen K,Ettwig K,Hu Z,Jetten M,Kartal B

    更新日期:2012-04-01 00:00:00

  • Listeria phage and phage tail induction triggered by components of bacterial growth media (phosphate, LiCl, nalidixic acid, and acriflavine).

    abstract::The detection of Listeria monocytogenes from food is currently carried out using a double enrichment. For the ISO methodology, this double enrichment is performed using half-Fraser and Fraser broths, in which the overgrowth of L. innocua can occur in samples where both species are present. In this study, we analyzed t...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.03235-14

    authors: Lemaître JP,Duroux A,Pimpie R,Duez JM,Milat ML

    更新日期:2015-03-01 00:00:00

  • Population dynamics of Bifidobacterium species in human feces during raffinose administration monitored by fluorescence in situ hybridization-flow cytometry.

    abstract::The population dynamics of bifidobacteria in human feces during raffinose administration were investigated at the species level by using fluorescence in situ hybridization (FISH) coupled with flow cytometry (FCM) analysis. Although double-staining FISH-FCM using both fluorescein isothiocyanate (FITC) and indodicarbocy...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01777-06

    authors: Dinoto A,Marques TM,Sakamoto K,Fukiya S,Watanabe J,Ito S,Yokota A

    更新日期:2006-12-01 00:00:00

  • Application of a competition model to the growth of Streptococcus mutans and Streptococcus sanguis in binary continuous culture.

    abstract::Streptococcus mutans 6715-15 and Streptococcus sanguis 10558 were grown together in continuous culture with glucose as the limiting carbon source. The relationship of growth rate to substrate concentration was determined for pure cultures of each organism in continuous and batch cultures. A model based on competition ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.45.4.1277-1282.1983

    authors: Kemp CW,Robrish SA,Curtis MA,Sharer SA,Bowen WH

    更新日期:1983-04-01 00:00:00

  • Extracellular Polysaccharide Is Not Responsible for Aluminum Tolerance of Rhizobium leguminosarum bv. Phaseoli CIAT899.

    abstract::Strain UHM-5, a pSym Exo derivative of the aluminum-tolerant Rhizobium leguminosarum bv. phaseoli strain CIAT899, was equally tolerant of aluminum (Al) as the parental culture. Dialyzed culture supernatants of the wild-type cells grown in YEM broth (10 cells ml) contained 185 mug of glucose equivalents ml whereas UHM-...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.58.4.1095-1101.1992

    authors: Kingsley MT,Bohlool BB

    更新日期:1992-04-01 00:00:00

  • Improved microautoradiographic method to determine individual microorganisms active in substrate uptake in natural waters.

    abstract::We report a method which combines epifluorescence microscopy and microautoradiography to determine both the total number of microorganisms in natural water populations and those individual organisms active in the uptake of specific substrates. After incubation with H-labeled substrate, the sample is filtered and, whil...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.44.4.945-953.1982

    authors: Tabor PS,Neihof RA

    更新日期:1982-10-01 00:00:00

  • Genotypic and phenotypic diversity within species of purple nonsulfur bacteria isolated from aquatic sediments.

    abstract::To assess the extent of genotypic and phenotypic diversity within species of purple nonsulfur bacteria found in aquatic sediments, a total of 128 strains were directly isolated from agar plates that had been inoculated with sediment samples from Haren and De Biesbosch in The Netherlands. All isolates were initially ch...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/aem.68.7.3467-3477.2002

    authors: Oda Y,Wanders W,Huisman LA,Meijer WG,Gottschal JC,Forney LJ

    更新日期:2002-07-01 00:00:00

  • Unique substrate spectrum and PCR application of Nanoarchaeum equitans family B DNA polymerase.

    abstract::The known archaeal family B DNA polymerases are unable to participate in the PCR in the presence of uracil. Here, we report on a novel archaeal family B DNA polymerase from Nanoarchaeum equitans that can successfully utilize deaminated bases such as uracil and hypoxanthine and on its application to PCR. N. equitans fa...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.00624-08

    authors: Choi JJ,Song JG,Nam KH,Lee JI,Bae H,Kim GA,Sun Y,Kwon ST

    更新日期:2008-11-01 00:00:00

  • Production of Ophthalmic Acid Using Engineered Escherichia coli.

    abstract::Ophthalmic acid (OA; l-γ-glutamyl-l-2-aminobutyryl-glycine) is an analog of glutathione (GSH; l-γ-glutamyl-l-cysteinyl-glycine) in which the cysteine moiety is replaced by l-2-aminobutyrate. OA is a useful peptide for the pharmaceutical and/or food industries. Herein, we report a method for the production of OA using ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.02806-17

    authors: Ito T,Tokoro M,Hori R,Hemmi H,Yoshimura T

    更新日期:2018-03-19 00:00:00

  • Application of a specific and sensitive radiometric assay for microbial lipase activities in marine water samples from the lagoon of nouméa.

    abstract::Marine microbiologists commonly assay lipase activities by using a synthetic fluorescent analog, 4-methylumbelliferyl (MUF)-oleate. The technique is convenient, but it is considered to be unspecific because of the structure of this analog. This study reports the design of a new specific and sensitive lipase assay base...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/aem.69.12.7395-7400.2003

    authors: Bourguet N,Torréton JP,Galy O,Arondel V,Goutx M

    更新日期:2003-12-01 00:00:00

  • Engineering of bacterial methyl ketone synthesis for biofuels.

    abstract::We have engineered Escherichia coli to overproduce saturated and monounsaturated aliphatic methyl ketones in the C₁₁ to C₁₅ (diesel) range; this group of methyl ketones includes 2-undecanone and 2-tridecanone, which are of importance to the flavor and fragrance industry and also have favorable cetane numbers (as we re...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.06785-11

    authors: Goh EB,Baidoo EE,Keasling JD,Beller HR

    更新日期:2012-01-01 00:00:00

  • Mycotoxins produced from fungi isolated from foodstuffs and soil: comparison of toxicity in fibroblasts and rat feeding tests.

    abstract::Thirty-nine isolates of fungi obtained from foodstuffs and soil samples from various parts of the world have been identified. The isolates were grown on a solid rice medium, and extracts were prepared with 50% aqueous methanol. The extracts were examined for toxicity in the following systems: (i) cytotoxicity to cultu...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.48.3.654-661.1984

    authors: Abbas HK,Mirocha CJ,Shier WT

    更新日期:1984-09-01 00:00:00

  • Quantification of yeast and bacterial gene transcripts in retail cheeses by reverse transcription-quantitative PCR.

    abstract::The cheese microbiota contributes to a large extent to the development of the typical color, flavor, and texture of the final product. Its composition is not well defined in most cases and varies from one cheese to another. The aim of the present study was to establish procedures for gene transcript quantification in ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.02360-12

    authors: Monnet C,Straub C,Castellote J,Onesime D,Bonnarme P,Irlinger F

    更新日期:2013-01-01 00:00:00

  • Role of host nutrition in symbiont regulation: impact of dietary nitrogen on proliferation of obligate and facultative bacterial endosymbionts of the pea aphid Acyrthosiphon pisum.

    abstract::The impact of host nutrition on symbiont regulation in the pea aphid Acyrthosiphon pisum was investigated. The population density of the obligate symbiont Buchnera aphidicola positively correlated with dietary nitrogen levels. In contrast, the population density of the facultative symbiont Serratia symbiotica increase...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01211-06

    authors: Wilkinson TL,Koga R,Fukatsu T

    更新日期:2007-02-01 00:00:00

  • Bioassay and dose measurement in UV disinfection.

    abstract::A bioassay method was developed to measure the average intensity within a UV disinfection reactor. The survival of spores of Bacillus subtilis was determined as a function of UV dose to prepare a standard curve. Spores were added to unknown systems, and the survival rate was used to determine the average intensity. A ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.45.3.872-877.1983

    authors: Qualls RG,Johnson JD

    更新日期:1983-03-01 00:00:00

  • Importance of polyphosphate kinase 1 for Campylobacter jejuni viable-but-nonculturable cell formation, natural transformation, and antimicrobial resistance.

    abstract::Campylobacter jejuni, a gram-negative, microaerophilic bacterium, is a predominant cause of bacterial gastroenteritis in humans. Although considered fragile and fastidious and lacking many classical stress response mechanisms, C. jejuni exhibits a remarkable capacity for survival and adaptation, successfully infecting...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.01603-09

    authors: Gangaiah D,Kassem II,Liu Z,Rajashekara G

    更新日期:2009-12-01 00:00:00

  • Broad-Host-Range Agrocin of Agrobacterium tumefaciens.

    abstract::Eighteen strains of Agrobacterium tumefaciens isolated from crown galls were tested for agrocin production. Of six agrocin-producing strains, one (D286) produced a broad-host-range agrocin active against strains carrying nopaline, octopine, and agropine type Ti plasmids. Sensitivity to agrocin D286 was found to map in...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.45.5.1526-1532.1983

    authors: Hendson M,Askjaer L,Thomson JA,van Montagu M

    更新日期:1983-05-01 00:00:00

  • Association of Campylobacter jejuni with laying hens and eggs.

    abstract::Laying hens were individually caged at 20 weeks of age and tested for fecal excretion of Campylobacter jejuni (minimum level of detection was 100 cfu/g) during a 42-week period. Peak rates of C. jejuni isolation (approximately 25% of hens positive) occurred at two different times, in October and in late April to early...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.47.3.533-536.1984

    authors: Doyle MP

    更新日期:1984-03-01 00:00:00

  • Metabolism of styrene by Rhodococcus rhodochrous NCIMB 13259.

    abstract::Rhodococcus rhodochrous NCIMB 13259 grows on styrene, toluene, ethylbenzene, and benzene as sole carbon sources. Simultaneous induction tests with cells grown on styrene or toluene showed high rates of oxygen consumption with toluene cis-glycol and 3-methylcatechol, suggesting the involvement of a cis-glycol pathway. ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.60.4.1137-1145.1994

    authors: Warhurst AM,Clarke KF,Hill RA,Holt RA,Fewson CA

    更新日期:1994-04-01 00:00:00

  • Field-based stable isotope probing reveals the identities of benzoic acid-metabolizing microorganisms and their in situ growth in agricultural soil.

    abstract::We used a combination of stable isotope probing (SIP), gas chromatography-mass spectrometry-based respiration, isolation/cultivation, and quantitative PCR procedures to discover the identity and in situ growth of soil microorganisms that metabolize benzoic acid. We added [(13)C]benzoic acid or [(12)C]benzoic acid (100...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.00464-08

    authors: Pumphrey GM,Madsen EL

    更新日期:2008-07-01 00:00:00

  • Thiosulfate-dependent chemolithoautotrophic growth of Bradyrhizobium japonicum.

    abstract::Thiosulfate-oxidizing sox gene homologues were found at four loci (I, II, III, and IV) on the genome of Bradyrhizobium japonicum USDA110, a symbiotic nitrogen-fixing bacterium in soil. In fact, B. japonicum USDA110 can oxidize thiosulfate and grow under a chemolithotrophic condition. The deletion mutation of the soxY(...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.02783-09

    authors: Masuda S,Eda S,Ikeda S,Mitsui H,Minamisawa K

    更新日期:2010-04-01 00:00:00

  • Respiratory Physiology of Lactococcus lactis in Chemostat Cultures and Its Effect on Cellular Robustness in Frozen and Freeze-Dried Starter Cultures.

    abstract::In this study, we used chemostat cultures to analyze the quantitative effects of the specific growth rate and respiration on the metabolism in Lactococcus lactis CHCC2862 and on the downstream robustness of cells after freezing or freeze-drying. Under anaerobic conditions, metabolism remained homofermentative, althoug...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.02785-19

    authors: Johanson A,Goel A,Olsson L,Franzén CJ

    更新日期:2020-03-02 00:00:00

  • Bacteria on closed-boll and commercially harvested cotton.

    abstract::The bacterial content of specially treated cottons used by other investigators to test human pulmonary responses to cotton dust was examined. Cotton from Lubbock, Tex. and Stoneville, Miss. were either (i) harvested by machine and handled as commercial bale cotton, (ii) harvested as closed bolls with bracts intact and...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章

    doi:10.1128/AEM.44.2.355-362.1982

    authors: Millner PD,Ericson KE,Marsh PB

    更新日期:1982-08-01 00:00:00