High-level extracellular expression of κ-carrageenase in Brevibacillus choshinensis for the production of a series of κ-carrageenan oligosaccharides

2018 ◽  
Vol 64 ◽  
pp. 83-92 ◽  
Author(s):  
Yujuan Zhao ◽  
Zhe Chi ◽  
Yanyan Xu ◽  
Nianci Shi ◽  
Zhenming Chi ◽  
...  
Keyword(s):  
Extracellular Expression ◽  
Brevibacillus Choshinensis ◽  
High Level

scholarly journals Constitutive High Level Expression of an Endoxylanase Gene from the Newly IsolatedBacillus subtilisAQ1 inEscherichia coli

2010 ◽  
Vol 2010 ◽  
pp. 1-12 ◽  
Author(s):  
Is Helianti ◽  
Niknik Nurhayati ◽  
Maria Ulfah ◽  
Budiasih Wahyuntari ◽  
Siswa Setyahadi
Keyword(s):  
Specific Activity ◽  
Biochemical Properties ◽  
Extracellular Expression ◽  
High Level Expression ◽  
Periplasmic Fraction ◽  
16S Rdna Sequence ◽  
E Coli ◽  
Encoding Gene ◽  
High Level ◽  
Very High

A xylanolytic bacterium was isolated from the sediment of an aquarium. Based on the 16S rDNA sequence as well as morphological and biochemical properties the isolate was identified and denoted asBacillus subtilis(B. subtilis) AQ1 strain. An endoxylanase-encoding gene along with its indigenous promoter was PCR amplified and after cloning expressed inE. coli. InE. colithe recombinant enzyme was found in the extracellular, in the cytoplasmic, and in the periplasmic fraction. The specific activity of the extracellular AQ1 recombinant endoxylanase after 24-hour fermentation was very high, namely,2173.6 ± 51.4and2745.3 ± 11 U/mg in LB and LB-xylan medium, respectively. This activity was clearly exceeding that of the nativeB. subtilisAQ1 endoxylanase and that of 95% homologous recombinant one fromB. subtilisDB104. The result shows that the original AQ1 endoxylanase promoter and the signal peptide gave a very high constitutive extracellular expression inE. coliand hence made the production inE. colifeasible.

scholarly journals Extracellular expression of Aerococcus viridans pyruvate oxidase in recombinant Escherichia coli through SecB co-expression

RSC Advances ◽  
2019 ◽  
Vol 9 (45) ◽  
pp. 26291-26301 ◽  
Author(s):  
Junwen Lu ◽  
Jianguo Zhang
Keyword(s):  
Escherichia Coli ◽  
Recombinant Escherichia Coli ◽  
Extracellular Expression ◽  
Pyruvate Oxidase ◽  
E Coli ◽  
Aerococcus Viridans ◽  
High Level

Extracellular pyruvate oxidase was expressed at a high level using E. coli by co-expression of chaperone SecB under bla promoter, and therefore cultivation optimization.

scholarly journals High-Level Extracellular Expression of a New β-N-Acetylglucosaminidase in Escherichia coli for Producing GlcNAc

2021 ◽  
Vol 12 ◽  
Author(s):  
Congna Li ◽  
Shun Jiang ◽  
Chao Du ◽  
Zhenghui Lu ◽  
Nisha He ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Chemical Process ◽  
Specific Activity ◽  
High Specific Activity ◽  
Culture Conditions ◽  
Extracellular Expression ◽  
Secreted Expression ◽  
Encoding Gene ◽  
High Level ◽  
The Cost

N-acetyl-β-D glucosamine (GlcNAc) is wildly used in cosmetics, nutraceuticals and pharmaceuticals. The traditional chemical process for GlcNAc production from chitin causes serious acidic pollution. Therefore, the enzymatic hydrolysis becomes a great promising and alternative strategy to produce GlcNAc. β-N-acetylglucosaminidase (NAGase) can hydrolyze chitin to produce GlcNAc. Here, a GH3 family NAGase encoding gene BlNagZ from Bacillus licheniformis was expressed extracellularly in Escherichia coli guided by signal peptide PelB. The recombinant BlNagZ presented the best activity at 60°C and pH 5.5 with a high specific activity of 13.05 U/mg. The BlNagZ activity in the fermentation supernatant can reach 13.62 U/mL after optimizing the culture conditions, which is 4.25 times higher than optimization before. Finally, combining BlNagZ with chitinase ChiA we identified before, chitin conversion efficiency to GlcNAc can reach 89.2% within 3.5 h. In all, this study provided not only a high active NAGase, and a secreted expression strategy to reduce the cost of production, which is conducive to the industrial application.

High-Level Production of Recombinant Chicken Interferon-γ by Brevibacillus choshinensis

2001 ◽  
Vol 23 (1) ◽  
pp. 113-120 ◽  
Author(s):  
Koji Yashiro ◽  
John W. Lowenthal ◽  
Terri E. O'Neil ◽  
Shogo Ebisu ◽  
Hiroaki Takagi ◽  
...  
Keyword(s):  
Interferon Γ ◽  
Brevibacillus Choshinensis ◽  
High Level ◽  
Level Production

scholarly journals High-level intracellular expression of heterologous proteins in Brevibacillus choshinensis SP3 under the control of a xylose inducible promoter

2013 ◽  
Vol 12 (1) ◽  
pp. 12 ◽  
Author(s):  
Nunzia D’Urzo ◽  
Manuele Martinelli ◽  
Chiara Nenci ◽  
Cecilia Brettoni ◽  
John L Telford ◽  
...  
Keyword(s):  
Inducible Promoter ◽  
Heterologous Proteins ◽  
Intracellular Expression ◽  
Brevibacillus Choshinensis ◽  
High Level

High-level extracellular expression of inulin fructotransferase in Pichia pastoris for DFA III production

2014 ◽  
Vol 95 (7) ◽  
pp. 1408-1413 ◽  
Author(s):  
Rongrong Zhan ◽  
Wanmeng Mu ◽  
Bo Jiang ◽  
Yungao Li ◽  
Liuming Zhou ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Extracellular Expression ◽  
Inulin Fructotransferase ◽  
High Level ◽  
Dfa Iii

Transcription factor/DNA interactions visualized by electron spectroscopic imaging

1992 ◽  
Vol 50 (1) ◽  
pp. 450-451
Author(s):  
David P. Bazett-Jones ◽  
Mark L. Brown
Keyword(s):  
Transcription Factor ◽  
Dna Sequences ◽  
Transcription Initiation ◽  
Molecular Mechanisms ◽  
Phosphorus Content ◽  
Spectroscopic Imaging ◽  
Electron Spectroscopic Imaging ◽  
Dna Backbone ◽  
Two Parameters ◽  
High Level

A multisubunit RNA polymerase enzyme is ultimately responsible for transcription initiation and elongation of RNA, but recognition of the proper start site by the enzyme is regulated by general, temporal and gene-specific trans-factors interacting at promoter and enhancer DNA sequences. To understand the molecular mechanisms which precisely regulate the transcription initiation event, it is crucial to elucidate the structure of the transcription factor/DNA complexes involved. Electron spectroscopic imaging (ESI) provides the opportunity to visualize individual DNA molecules. Enhancement of DNA contrast with ESI is accomplished by imaging with electrons that have interacted with inner shell electrons of phosphorus in the DNA backbone. Phosphorus detection at this intermediately high level of resolution (≈lnm) permits selective imaging of the DNA, to determine whether the protein factors compact, bend or wrap the DNA. Simultaneously, mass analysis and phosphorus content can be measured quantitatively, using adjacent DNA or tobacco mosaic virus (TMV) as mass and phosphorus standards. These two parameters provide stoichiometric information relating the ratios of protein:DNA content.

Advances in Stem Instrumentation for Biology

1990 ◽  
Vol 48 (1) ◽  
pp. 362-363
Author(s):  
J. S. Wall
Keyword(s):  
Scanning Transmission Electron Microscope ◽  
Carbon Films ◽  
Specimen Preparation ◽  
Material Deposition ◽  
Active User ◽  
Percent Improvement ◽  
Scanning Transmission ◽  
The Moment ◽  
Film Substrate ◽  
High Level

The forte of the Scanning transmission Electron Microscope (STEM) is high resolution imaging with high contrast on thin specimens, as demonstrated by visualization of single heavy atoms. of equal importance for biology is the efficient utilization of all available signals, permitting low dose imaging of unstained single molecules such as DNA.Our work at Brookhaven has concentrated on: 1) design and construction of instruments optimized for a narrow range of biological applications and 2) use of such instruments in a very active user/collaborator program. Therefore our program is highly interactive with a strong emphasis on producing results which are interpretable with a high level of confidence.The major challenge we face at the moment is specimen preparation. The resolution of the STEM is better than 2.5 A, but measurements of resolution vs. dose level off at a resolution of 20 A at a dose of 10 el/A2 on a well-behaved biological specimen such as TMV (tobacco mosaic virus). To track down this problem we are examining all aspects of specimen preparation: purification of biological material, deposition on the thin film substrate, washing, fast freezing and freeze drying. As we attempt to improve our equipment/technique, we use image analysis of TMV internal controls included in all STEM samples as a monitor sensitive enough to detect even a few percent improvement. For delicate specimens, carbon films can be very harsh-leading to disruption of the sample. Therefore we are developing conducting polymer films as alternative substrates, as described elsewhere in these Proceedings. For specimen preparation studies, we have identified (from our user/collaborator program ) a variety of “canary” specimens, each uniquely sensitive to one particular aspect of sample preparation, so we can attempt to separate the variables involved.

A Questionnaire Survey of Current Rehabilitation Practices for Adults With Normal Hearing Sensitivity Who Experience Auditory Difficulties

2020 ◽  
Vol 29 (4) ◽  
pp. 738-761
Author(s):  
Tess K. Koerner ◽  
Melissa A. Papesh ◽  
Frederick J. Gallun
Keyword(s):  
Hearing Aids ◽  
Questionnaire Survey ◽  
Patient Population ◽  
Clinical Care ◽  
Normal Hearing ◽  
Auditory Training ◽  
Training Methods ◽  
Hearing Sensitivity ◽  
Considerable Benefit ◽  
High Level

Purpose A questionnaire survey was conducted to collect information from clinical audiologists about rehabilitation options for adult patients who report significant auditory difficulties despite having normal or near-normal hearing sensitivity. This work aimed to provide more information about what audiologists are currently doing in the clinic to manage auditory difficulties in this patient population and their views on the efficacy of recommended rehabilitation methods. Method A questionnaire survey containing multiple-choice and open-ended questions was developed and disseminated online. Invitations to participate were delivered via e-mail listservs and through business cards provided at annual audiology conferences. All responses were anonymous at the time of data collection. Results Responses were collected from 209 participants. The majority of participants reported seeing at least one normal-hearing patient per month who reported significant communication difficulties. However, few respondents indicated that their location had specific protocols for the treatment of these patients. Counseling was reported as the most frequent rehabilitation method, but results revealed that audiologists across various work settings are also successfully starting to fit patients with mild-gain hearing aids. Responses indicated that patient compliance with computer-based auditory training methods was regarded as low, with patients generally preferring device-based rehabilitation options. Conclusions Results from this questionnaire survey strongly suggest that audiologists frequently see normal-hearing patients who report auditory difficulties, but that few clinicians are equipped with established protocols for diagnosis and management. While many feel that mild-gain hearing aids provide considerable benefit for these patients, very little research has been conducted to date to support the use of hearing aids or other rehabilitation options for this unique patient population. This study reveals the critical need for additional research to establish evidence-based practice guidelines that will empower clinicians to provide a high level of clinical care and effective rehabilitation strategies to these patients.

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