Production enhancement of the extracellular lipase LipA in Bacillus subtilis: Effects of expression system and Sec pathway components

2018 ◽  
Vol 142 ◽  
pp. 81-87 ◽  
Author(s):  
Ran Jing Ma ◽  
Yan Hong Wang ◽  
Lu Liu ◽  
Lei Lei Bai ◽  
Rui Ban
Keyword(s):  
Bacillus Subtilis ◽  
Expression System ◽  
Extracellular Lipase ◽  
Sec Pathway

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.

scholarly journals Cell Membrane-Interrupting Antimicrobial Peptides from Isatis indigotica Fortune Isolated by a Bacillus subtilis Expression System

Biomolecules ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 30 ◽  
Author(s):  
Jia Wu ◽  
Hafiz Muhammad Khalid Abbas ◽  
Jiale Li ◽  
Yuan Yuan ◽  
Yunjun Liu ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Cell Membrane ◽  
Antimicrobial Peptides ◽  
Viral Infections ◽  
Clinical Medicine ◽  
Raw Materials ◽  
Expression System ◽  
Electrical Potential ◽  
Antimicrobial Activities ◽  
Isatis Indigotica

The situation of drug resistance has become more complicated due to the scarcity of plant resistance genes, and overcoming this challenge is imperative. Isatis indigotica has been used for the treatment of wounds, viral infections, and inflammation for centuries. Antimicrobial peptides (AMPs) are found in all classes of life ranging from prokaryotes to eukaryotes. To identify AMPs, I. indigotica was explored using a novel, sensitive, and high-throughput Bacillus subtilis screening system. We found that IiR515 and IiR915 exhibited significant antimicrobial activities against a variety of bacterial (Xanthomonas oryzae, Ralstonia solanacearum, Clavibacter michiganensis, and C. fangii) and fungal (Phytophthora capsici and Botrytis cinerea) pathogens. Scanning electron microscope and cytometric analysis revealed the possible mechanism of these peptides, which was to target and disrupt the bacterial cell membrane. This model was also supported by membrane fluidity and electrical potential analyses. Hemolytic activity assays revealed that these peptides may act as a potential source for clinical medicine development. In conclusion, the plant-derived novel AMPs IiR515 and IiR915 are effective biocontrol agents and can be used as raw materials in the drug discovery field.

scholarly journals Precise Deletion of tagD and Controlled Depletion of Its Product, Glycerol 3-Phosphate Cytidylyltransferase, Leads to Irregular Morphology and Lysis of Bacillus subtilisGrown at Physiological Temperature

2001 ◽  
Vol 183 (22) ◽  
pp. 6688-6693 ◽  
Author(s):  
Amit P. Bhavsar ◽  
Terry J. Beveridge ◽  
Eric D. Brown
Keyword(s):  
Bacillus Subtilis ◽  
Gene Product ◽  
Genetic Background ◽  
Expression System ◽  
Teichoic Acid ◽  
Structural Defects ◽  
Acid Biosynthesis ◽  
Physiological Temperature ◽  
Conditional Expression ◽  
Phenotypic Rescue

ABSTRACT Using a previously reported conditional expression system for use in Bacillus subtilis (A. P. Bhavsar, X. Zhao, and E. D. Brown, Appl. Environ. Microbiol. 67:403–410, 2001), we report the first precise deletion of a teichoic acid biosynthesis (tag) gene, tagD,in B. subtilis. This teichoic acid mutant showed a lethal phenotype when characterized at a physiological temperature and in a defined genetic background. This tagD mutant was subject to full phenotypic rescue upon expression of the complementing copy oftagD. Depletion of the tagD gene product (glycerol 3-phosphate cytidylyltransferase) via modulated expression of tagD from the amyE locus revealed structural defects centered on shape, septation, and division. Thickening of the wall and ultimately lysis followed these events.

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