scholarly journals Construction of a Mutant Bacillus Subtilis Strain for High Purity Poly-γ-Glutamic Acid Production

2022 ◽  
Author(s):  
Linlin He ◽  
Lu Liu ◽  
Rui Ban
Keyword(s):  
Bacillus Subtilis ◽  
Glutamic Acid ◽  
Protein Content ◽  
Protein Purification ◽  
Drug Carrier ◽  
Specific Protein ◽  
Extracellular Protein ◽  
Subtilis Strain ◽  
Potential Applications ◽  
Glutamic Acid Production

Abstract Purpose To construct a Bacillus subtilis strain for improved purity of poly-γ-glutamic acid. Results The construction of strain GH16 was achieved by knocking out five extracellular protein genes and an operon from Bacillus subtilis G423. Then we analyzed the protein content in the γ-PGA produced by the resultant strain GH16/pHPG which decreased by 6.08%. Subsequently the fla-che operon, PBSX and the yrpD, ywoF and yclQ genes were knocked out successively and the mutant strain GH17, GH18, and GH19 was obtained. Ultimately, the protein content was reduced by 43.9%. In addition, the polysaccharide content in the γ-PGA was decreased from 2.21–1.93% due to the epsA-O operon was knocked. Conclusion γ-PGA has potential applications as a drug carrier, sustained-releasing agent and medical composite in medicine. To our knowledge, this is the first report of engineered Bacillus subtilis strains which can produce γ-PGA with a purity higher than 97%. Our results confirmed that this upstream strategy significantly enhanced specific protein purity by the removal of extracellular protein genes in Bacillus subtilis, and it is promising in other protein purification.

scholarly journals Regulation of poly-γ-glutamic acid production in Bacillus subtilis GXA-28 by potassium

2016 ◽  
Vol 61 ◽  
pp. 83-89 ◽  
Author(s):  
Wei Zeng ◽  
Zhiqun Liang ◽  
Zhi Li ◽  
Yaxi Bian ◽  
Zhihong Li ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Glutamic Acid ◽  
Acid Production ◽  
Glutamic Acid Production

scholarly journals Poly-l-gamma-glutamic acid production by recombinant Bacillus subtilis without pgsA gene

AMB Express ◽  
2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Kazuhisa Sawada ◽  
Hiroyuki Araki ◽  
Yasushi Takimura ◽  
Kenta Masuda ◽  
Yasushi Kageyama ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Glutamic Acid ◽  
Acid Production ◽  
Glutamic Acid Production ◽  
Recombinant Bacillus Subtilis

scholarly journals Complete Genome Sequence of Bacillus subtilis Strain DKU_NT_02, Isolated from Traditional Korean Food Using Soybean (Chung-gook-jang) for High-Quality Poly-γ-Glutamic Acid Activity

2018 ◽  
Vol 6 (25) ◽  
Author(s):  
Man-Seok Bang ◽  
Hee-Won Jeong ◽  
Yea-jin Lee ◽  
Su Ji Lee ◽  
Sang-Cheol Lee ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Glutamic Acid ◽  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Genetic Factors ◽  
Subtilis Strain ◽  
High Quality ◽  
Content Type ◽  
Korean Food

The complete genome sequence of Bacillus subtilis strain DKU_NT_02, isolated from traditional Korean food using soybeans (chung-gook-jang), is presented here. This strain was chosen to help identify genetic factors with high-quality poly-γ-glutamic acid (γPGA) activity.

scholarly journals The effect of magnesium ion deprivation on the synthesis of mucopeptide and its precursors in Bacillus subtilis

1969 ◽  
Vol 115 (3) ◽  
pp. 419-430 ◽  
Author(s):  
A J Garrett
Keyword(s):  
Bacillus Subtilis ◽  
Glutamic Acid ◽  
Trichloroacetic Acid ◽  
Amino Sugar ◽  
Diaminopimelic Acid ◽  
Magnesium Ion ◽  
Subtilis Strain ◽  
Amino Sugars ◽  
Intracellular Concentrations

1. Mg2+ or Mn2+ starvation causes suspensions of Bacillus subtilis strain W 23 to accumulate bound amino sugars that are soluble in trichloroacetic acid. 2. The presence of chloramphenicol or puromycin produces higher intracellular concentrations of amino sugars during Mg2+ starvation, but neither compound can stimulate the accumulation when Mg2+ is present. 3. The major component of the amino sugar fraction extracted from cells deprived of Mg2+ is a nucleotide containing uridine, phosphorus, N-acetylmuramic acid, alanine, glutamic acid and α∈-diaminopimelic acid in the molar proportions of 1:2:1:3:1:1. This compound represents at least 80% of the bound N-acetylhexosamine extracted by trichloroacetic acid. 4. Studies of the binding of this nucleotide with vancomycin support the proposal that it is the mucopeptide precursor UDP-N-acetylmuramyl-l-alanyl-d-glutaminyl- α∈-diaminopimelyl-d-alanyl-d-alanine. 5. A method is described for the isolation of this material labelled with [3H]α∈-diaminopimelic acid. 6. When Mg2+ is supplied to cells previously starved of Mg2+, the accumulated pool of amino sugars rapidly decreases. 7. The biosynthesis of mucopeptide is inhibited by 35–50% under conditions of Mg2+ starvation. The presence of EDTA increases this inhibition to 70%. The amount of N-acetylhexosamine that accumulates is balanced exactly by the associated fall in mucopeptide synthesis. 8. ‘Chase’ experiments show that the accumulated N-acetylhexosamine compound is utilized in mucopeptide synthesis.

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