Construction of Chromosomally Located T7 Expression System for Production of Heterologous Secreted Proteins in Bacillus subtilis

2010 ◽  
Vol 58 (9) ◽  
pp. 5392-5399 ◽  
Author(s):  
Po Ting Chen ◽  
Jei-Fu Shaw ◽  
Yun-Peng Chao ◽  
Tuan-Hua David Ho ◽  
Su-May Yu
Keyword(s):  
Bacillus Subtilis ◽  
Expression System ◽  
Secreted Proteins ◽  
T7 Expression System

scholarly journals Using cellulose binding module of CBM3A as the purification tag for secreted Endoglucanase A (CelA) in Bacillus subtilis

2016 ◽  
Vol 19 (2) ◽  
pp. 5-14
Author(s):  
Phuong Hoang Ngoc Nguyen ◽  
Phuoc Thanh Nguyen ◽  
Thang Luong Pham ◽  
Hoang Duc Nguyen ◽  
Thuoc Linh Tran ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Protein Purification ◽  
Clostridium Thermocellum ◽  
Expression System ◽  
Secreted Proteins ◽  
Amorphous Cellulose ◽  
Recombinant Protein Purification ◽  
Bacillus Subtilis Wb800n ◽  
Cellulose Binding

Traditional methods of recombinant protein purification are uneconomic and inconvenient to the secreted proteins at large-volume. CBM3a, a module from cellulosome’s scaffoldin of Clostridium thermocellum, directs the binding of the cellulase complex on the cheap cellulose substrate. Most of previous studies about CBM3a fused with cellulases as the purification tag were conducted in intracellular Escherichia coli system. In this research, we used the extracellular Bacillus subtilis WB800N expression system to investigate the CBM3a-tag fused with endoglucanase CelA into plasmid pHT. The results indicated that protein CelA was secrected and purified by CBM3a-tag binding on the Regenerated Amorphous Cellulose (RAC) subtrate. This can be used for further improvement in protein purification tag designing.

Production of HIV-1 gp120 in Packed-Bed Bioreactor Using the Vaccinia Virus/T7 Expression System

2000 ◽  
Vol 16 (5) ◽  
pp. 744-750 ◽  
Author(s):  
Y.-C. Hu ◽  
J. Kaufman ◽  
M.W. Cho ◽  
H. Golding ◽  
J. Shiloach
Keyword(s):  
Vaccinia Virus ◽  
Expression System ◽  
Packed Bed ◽  
Packed Bed Bioreactor ◽  
T7 Expression System ◽  
Hiv 1

scholarly journals Antimicrobial genes from Allium sativum and Pinellia ternata revealed by a Bacillus subtilis expression system

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Xi Kong ◽  
Mei Yang ◽  
Hafiz Muhammad Khalid Abbas ◽  
Jia Wu ◽  
Mengge Li ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Allium Sativum ◽  
Expression System ◽  
Pinellia Ternata ◽  
Bacillus Subtilis Expression

Production of Lysinibacillus sphaericus Mosquitocidal Protein Mtx2 from Bacillus subtilis as a Secretory Protein

2021 ◽  
Vol 28 ◽  
Author(s):  
Chutchanun Trakulnaleamsai ◽  
Boonhiang Promdonkoy ◽  
Sumarin Soonsanga
Keyword(s):  
Bacillus Subtilis ◽  
Larvicidal Activity ◽  
Mosquito Control ◽  
Expression System ◽  
Secretory Protein ◽  
Signal Peptides ◽  
Secreted Protein ◽  
Lysinibacillus Sphaericus ◽  
Putative Signal Peptide ◽  
Set Up

Background: Mtx2 is a mosquitocidal toxin produced during the vegetative growth of Lysinibacillus sphaericus. The protein shows synergism with other toxins against mosquito larvae; hence it could be used in mosquito control formulations. The protein expression system is needed for Mtx2 development as a biocontrol agent. Objective: The objective of the study was to set up a Bacillus subtilis system to produce Mtx2 as a secreted protein since the protein contains a putative signal peptide. Methods: Initially, four different promoters (P43, Pspac, PxylA, and PyxiE) were compared for their strength using GFP as a reporter in B. subtilis. Subsequently, six different signal peptides (SacB, Epr, AmyE, AprE, LipA, and Vip3A)were tested in conjunction with the selected promoter and mtx2 to evaluate levels of Mtx2 secreted by B. subtilis WB800, an extracellular protease-deficient strain. Results: The promoter PyxiE showed the highest GFP intensity and was selected for further study. Mtx2 was successfully produced as a secreted protein from signal peptides LipA and AmyE, and exhibited larvicidal activity against Aedesaegypti. Conclusion: B. subtilis was successfully developed as a host for the production of secreted Mtx2 and the protein retained its larvicidal activity. Although the Mtx2 production level still needs improvement, the constructed plasmids could be used to produce other soluble proteins.

Stable episomal expression system under control of a stress inducible promoter enhances the immunogenicity of Bacillus subtilis as a vector for antigen delivery

Vaccine ◽  
2006 ◽  
Vol 24 (15) ◽  
pp. 2935-2943 ◽  
Author(s):  
Juliano D. Paccez ◽  
Wilson B. Luiz ◽  
Maria E. Sbrogio-Almeida ◽  
Rita C.C. Ferreira ◽  
Wolfgang Schumann ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Expression System ◽  
Inducible Promoter ◽  
Antigen Delivery

scholarly journals Isolation of Antimicrobial Genes from Oryza rufipogon Griff by Using a Bacillus subtilis Expression System with Potential Antimicrobial Activities

2020 ◽  
Vol 21 (22) ◽  
pp. 8722
Author(s):  
Jiale Li ◽  
Samiul Islam ◽  
Pengfei Guo ◽  
Xiaoqing Hu ◽  
Wubei Dong
Keyword(s):  
Electron Microscopy ◽  
Bacillus Subtilis ◽  
Antimicrobial Peptides ◽  
Expression System ◽  
Oryza Rufipogon ◽  
Host Cells ◽  
Cdna Libraries ◽  
Potential Candidate ◽  
Oryza Rufipogon Griff ◽  
Bacillus Subtilis Expression

Antimicrobial genes are distributed in all forms of life and provide a primary defensive shield due to their unique broad-spectrum resistance activities. To better isolate these genes, we used the Bacillus subtilis expression system as the host cells to build Oryza rufipogon Griff cDNA libraries and screen potential candidate genes from the library at higher flux using built-in indicator bacteria. We observed that the antimicrobial peptides OrR214 and OrR935 have strong antimicrobial activity against a variety of Gram-positive and Gram-negative bacteria, as well as several fungal pathogens. Owing to their high thermal and enzymatic stabilities, these two peptides can also be used as field biocontrol agents. Furthermore, we also found that the peptide OrR214 (MIC 7.7–10.7 μM) can strongly inhibit bacterial growth compared to polymyxin B (MIC 5–25 μM) and OrR935 (MIC 33–44 μM). The cell flow analysis, reactive oxygen burst, and electron microscopy (scanning and transmission electron microscopy) observations showed that the cell membranes were targeted by peptides OrR214 and OrR935, which revealed the mode of action of bacteriostasis. Moreover, the hemolytic activity, toxicity, and salt sensitivity experiments demonstrated that these two peptides might have the potential to be used for clinical applications. Overall, OrR214 and OrR935 antimicrobial peptides have a high-throughput bacteriostatic activity that acts as a new form of antimicrobial agent and can be used as a raw material in the field of drug development.

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