Competence related proteins in the supernatant of competent cells of Bacillus subtilis

1985 ◽  
Vol 198 (2) ◽  
pp. 329-335 ◽  
Author(s):  
Charles W. Finn ◽  
Otto E. Landman
Keyword(s):  
Bacillus Subtilis ◽  
Competent Cells ◽  
Related Proteins

scholarly journals DNA-dependent ATPases in Bacillus subtilis Mutants and in Competent Cells

Microbiology ◽  
1984 ◽  
Vol 130 (1) ◽  
pp. 113-117
Author(s):  
G. Mazza ◽  
A. Sidoli ◽  
S. Riva
Keyword(s):  
Bacillus Subtilis ◽  
Competent Cells

scholarly journals Uptake of Synthetic Polynucleotides by Competent Cells of Bacillus subtilis

1970 ◽  
Vol 104 (2) ◽  
pp. 684-688 ◽  
Author(s):  
Orio Ciferri ◽  
Sergio Barlati ◽  
Joshua Lederberg
Keyword(s):  
Bacillus Subtilis ◽  
Competent Cells

scholarly journals Metabolic engineering of Bacillus subtilis with an Endopolygalacturonase gene Isolated from Pectobacterium carotovorum; a Plant Pathogenic bacterial strain.

2021 ◽  
Author(s):  
Nagina Rafique ◽  
Saiqa Bashir ◽  
Muhammad Zubair Khan ◽  
Imran Hayat ◽  
Willium Orts ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Enzyme Production ◽  
Metal Ion ◽  
Paper Industry ◽  
Expression System ◽  
Pathogenic Strain ◽  
Pectobacterium Carotovorum ◽  
Iptg Induction ◽  
Competent Cells ◽  
Bacillus Subtilis Wb800n

Pectinolytic enzymes [pectinases] produced by microbes are highly important for their biotechnological use in processing of vegetables and fruits beverages and use in pulp and paper industry. A pectinases, namely endo-polygalacturonase [endo-PGase], encoding gene isolated from Pectobacterium carotovorum, a plant pathogenic strain of bacteria was successfully cloned into a secretion vector pHT43 having σ?-dependent promoter P grac . For enhanced expression analysis, competent cells of Bacillus subtilis (WB800N) were prepared at stationary phase using high salt medium. The recombinant B. subtilis competent cells, harboring the engineered pHT43 with the endo-PGase gene were cultured in 2X-yeast extract tryptone medium. The recombinant endo-PGase enzyme was secreted directly into the medium after 72 hours of the first IPTG induction. The recombinant endo-PGase was screened for its activity at various temperatures and pH ranges. Optimal activity was found at pH 5.0 and a temperature of 40°C with a stability ranging from pH 5.0-9.0. For detection of metal ion effect, recombinant enzyme was incubated with 1mM concentration of; Ca ++ , Mg ++ , Zn ++ , EDTA, K ++ for 45 minutes. Resultantly, Ca ++ , EDTA and Zn ++ strongly inhibited the enzyme activity. The chromatographic analysis of enzymatic hydrolysate of polygalacturonic acid [PGA] and pectin substrates using HPLC and TLC revealed that tri and tetra-galacturonates were the end products of hydrolysis. The study led to the conclusion that endo-PGase gene from the plant pathogenic strain was successfully expressed in Bacillus subtilis and assessed for enzyme production using a very simple medium with IPTG induction. These findings proposed that the Bacillus expression system might be safe for commercial enzyme production as compared to yeast and fungi to escape endotoxins.

scholarly journals Bacillus subtilis YhaM, a Member of a New Family of 3′-to-5′ Exonucleases in Gram-Positive Bacteria

2002 ◽  
Vol 184 (22) ◽  
pp. 6250-6259 ◽  
Author(s):  
Irina A. Oussenko ◽  
Roberto Sanchez ◽  
David H. Bechhofer
Keyword(s):  
Bacillus Subtilis ◽  
Mrna Turnover ◽  
Wild Type ◽  
Rnase R ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
New Family ◽  
Double Stranded Dna ◽  
Ob Fold ◽  
Related Proteins

ABSTRACT A strain of Bacillus subtilis lacking two 3′-to-5′ exoribonucleases, polynucleotide phosphorylase (PNPase) and RNase R, was used to purify another 3′-to-5′ exoribonuclease, which is encoded by the yhaM gene. YhaM was active in the presence of Mn2+ (or Co2+), was inactive in the presence of Mg2+, and could also degrade single-stranded DNA. The half-life of bulk mRNA in a mutant lacking PNPase, RNase R, and YhaM was not significantly different from that of the wild type, suggesting the existence of additional activities that can participate in mRNA turnover. Sequence homologues of YhaM were found only in gram-positive organisms. The Staphylococcus aureus homologue, CBF1, which had been characterized as a double-stranded DNA binding protein involved in plasmid replication, was also shown to be an Mn2+-dependent exoribonuclease. YhaM protein has a C-terminal “HD domain,” found in metal-dependent phosphohydrolases. By structure modeling, it was shown that YhaM also contains an N-terminal “OB-fold,” present in many oligosaccharide- and oligonucleotide-binding proteins. The combination of these two domains is unique. Thus, YhaM and 10 related proteins from gram-positive organisms constitute a new exonuclease family.

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