Obtaining and purification of lysostaphin secreted in the culture liquid by Brevibacillus choshinensis/pNCMO2/lsf12 recombinant strain

2020 ◽  
Vol 36 (1) ◽  
pp. 7-15
Author(s):  
E.V. Baranova ◽  
V.V. Levchuk ◽  
T.V. Reshetnyak ◽  
P.V. Soloviev ◽  
N.A. Shishkova ◽  
...  
Keyword(s):  
Culture Medium ◽  
Expression System ◽  
Culture Liquid ◽  
Exchange Chromatography ◽  
High Yield ◽  
Diagnostic Tools ◽  
Yield Production ◽  
Brevibacillus Choshinensis ◽  
The Russian Federation ◽  
High Output

Here, «host-vector» expression system of Brevibacillus choshinensis was developed and used for producing a recombinant lysostaphin with high-output. The recombinant plasmid pNCMO2/lsf12 was constructed, and its expression in Brevibacillus choshinensis (strain Brevibacillus choshinensis/pNCMO2/lsf12) provided a synthesis of the 27-kDa protein, which was secreted into the culture medium. Its specific staphylolitic activity being 557 U/mg at optimal pH (7.5-8.0) and temperature (50-55 °C) values was comparable with the natural and recombinant analogs. We hope that developed methods of a deep cultivation of the recombinant Brevibacillus choshinensis/pNCMO2/lsf12 strain for a high-yield production (up to 90 mg/L) and a single-stage purification of lysostaphin (up to 90% homogeneity) become the basis for the production of the enzyme on an industrial scale. Brevibacillus choshinensis, ion-exchange chromatography, lysostaphin The work was financially supported by the Grant No. 050 of Rospotrebnadzor «Monitoring of borreliosis pathogens circulation in regions of the Russian Federation and improvement of diagnostic tools for borreliosis»

scholarly journals Efficient Expression of Maltohexaose-Forming α-Amylase from Bacillus stearothermophilus in Brevibacillus choshinensis SP3 and Its Use in Maltose Production

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Zhu Li ◽  
Lingqia Su ◽  
Xuguo Duan ◽  
Dan Wu ◽  
Jing Wu
Keyword(s):  
Culture Medium ◽  
Bacillus Stearothermophilus ◽  
Corn Starch ◽  
Expression System ◽  
Initial Ph ◽  
Brevibacillus Choshinensis ◽  
Efficient Expression ◽  
Cellular Integrity ◽  
Maltose Syrup ◽  
Starch Industry

The maltohexaose-forming, Ca2+-independent α-amylase gene from Bacillus stearothermophilus (AmyMH) was efficiently expressed in Brevibacillus choshinensis SP3. To improve the production of AmyMH in B. choshinensis SP3, the temperature and initial pH of culture medium were optimized. In addition, single-factor and response surface methodologies were pursued to optimize culture medium. Addition of proline to the culture medium significantly improved the production of recombinant α-amylase in B. choshinensis SP3. This improvement may result from improved cellular integrity of recombinant B. choshinensis SP3 in existence of proline. Culture medium optimization resulted in an 8-fold improvement in α-amylase yield, which reached 1.72 × 104 U·mL−1. The recombinant α-amylase was applied to the production of maltose on a laboratory scale. A maltose content of 90.72%, which could be classified as an extremely high maltose syrup, could be achieved using 15% (m/v) corn starch as the substrate. This study demonstrated that the B. choshinensis SP3 expression system was able to produce substantial quantities of recombinant α-amylase that has potential application in the starch industry.

High-yield production of a functional bacteriophage lysin with antipneumococcal activity using a plant virus-based expression system

2015 ◽  
Vol 200 ◽  
pp. 10-16 ◽  
Author(s):  
Urtė Starkevič ◽  
Luisa Bortesi ◽  
Marius Virgailis ◽  
Modestas Ružauskas ◽  
Anatoli Giritch ◽  
...  
Keyword(s):  
Plant Virus ◽  
Expression System ◽  
High Yield ◽  
Yield Production ◽  
High Yield Production

High-yield production of authentic human growth hormone using a plant virus-based expression system

2005 ◽  
Vol 3 (6) ◽  
pp. 613-620 ◽  
Author(s):  
Mario Gils ◽  
Romy Kandzia ◽  
Sylvestre Marillonnet ◽  
Victor Klimyuk ◽  
Yuri Gleba
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Plant Virus ◽  
Expression System ◽  
Human Growth ◽  
High Yield ◽  
Yield Production ◽  
High Yield Production

Random knock-in expression system for high yield production of heterologous protein in Bacillus subtilis

2018 ◽  
Vol 266 ◽  
pp. 50-58 ◽  
Author(s):  
Da-Eun Jeong ◽  
Younju So ◽  
Soo-Young Park ◽  
Seung-Hwan Park ◽  
Soo-Keun Choi
Keyword(s):  
Bacillus Subtilis ◽  
Heterologous Protein ◽  
Expression System ◽  
High Yield ◽  
Yield Production ◽  
High Yield Production

Evaluation of the Efficiency of Protein A Affinity Chromatography to Purify a Monoclonal Antibody for Cancer Treatment and its Purity Analysis

2020 ◽  
Vol 7 (2) ◽  
pp. 121-133
Author(s):  
Ayesha Akhtar ◽  
Shivakumar Arumugam ◽  
Shoaib Alam
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Affinity Chromatography ◽  
Protein A ◽  
Exchange Chromatography ◽  
Size Exclusion ◽  
High Yield ◽  
Staphylococcal Protein A ◽  
Purification Step ◽  
Purity Analysis

Background:: Protein A affinity chromatography is often employed as the most crucial purification step for monoclonal antibodies to achieve high yield with purity and throughput requirements. Introduction:: Protein A, also known as Staphylococcal protein A (SPA) is found in the cell wall of the bacteria staphylococcus aureus. It is one of the first discovered immunoglobulin binding molecules and has been extensively studied since the past few decades. The efficiency of Protein A affinity chromatography to purify a recombinant monoclonal antibody in a cell culture sample has been evaluated, which removes 99.0% of feed stream impurities. Materials and Method:: We have systematically evaluated the purification performance by using a battery of analytical methods SDS-PAGE (non-reduced and reduced sample), Cation Exchange Chromatography (CEX), Size-exclusion chromatography (SEC), and Reversed phased-Reduced Chromatography for a CHO-derived monoclonal antibody. Results and Discussion:: The analytical test was conducted to determine the impurity parameter, Host Cell Contaminating Proteins (HCP). It was evaluated to be 0.015ng/ml after the purification step; while initially, it was found to be 24.431ng/ml. Conclusion:: The tests showed a distinct decrease in the level of different impurities after the chromatography step. It can be concluded that Protein A chromatography is an efficient step in the purification of monoclonal antibodies.

scholarly journals Review: Managing sheep and goats for sustainable high yield production

animal ◽  
2021 ◽  
pp. 100293
Author(s):  
J. Simões ◽  
J.A. Abecia ◽  
A. Cannas ◽  
J.A. Delgadillo ◽  
D. Lacasta ◽  
...  
Keyword(s):  
High Yield ◽  
Sheep And Goats ◽  
Yield Production ◽  
High Yield Production

scholarly journals High‐Yield Production, Characterization, and Functionalization of Recombinant Magnetosomes in the Synthetic Bacterium Rhodospirillum rubrum “magneticum”

2021 ◽  
pp. 2101017
Author(s):  
Frank Mickoleit ◽  
Sabine Rosenfeldt ◽  
Mauricio Toro‐Nahuelpan ◽  
Miroslava Schaffer ◽  
Anna S. Schenk ◽  
...  
Keyword(s):  
Rhodospirillum Rubrum ◽  
High Yield ◽  
Yield Production ◽  
High Yield Production
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