In vivo functional expression of an extracellular Ca2+-independent Bacillus pumilus lipase in Bacillus subtilis WB800N

2015 ◽  
Vol 65 (4) ◽  
pp. 1973-1983 ◽  
Author(s):  
Jing Ma ◽  
Yushu Ma ◽  
Wei Wei ◽  
Dong-zhi Wei
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Pumilus ◽  
Functional Expression ◽  
Bacillus Subtilis Wb800n

scholarly journals Transcriptional Control of the Low-Temperature-Inducible des Gene, Encoding the Δ5 Desaturase of Bacillus subtilis

1999 ◽  
Vol 181 (22) ◽  
pp. 7028-7033 ◽  
Author(s):  
Pablo S. Aguilar ◽  
Paloma Lopez ◽  
Diego de Mendoza
Keyword(s):  
Bacillus Subtilis ◽  
Transcriptional Control ◽  
Unsaturated Fatty Acids ◽  
Functional Expression ◽  
Mrna Levels ◽  
Gene Encoding ◽  
Major Mechanism ◽  
Temperature Downshift ◽  
The Stability

ABSTRACT The Bacillus subtilis des gene encodes the cold-inducible Δ5 lipid desaturase involved in the formation of unsaturated fatty acids from saturated phospholipid precursors. Here, we describe the expression pattern of the des gene in response to a temperature downshift from 37 to 20°C. We found that the synthesis of des mRNA is undetectable at 37°C but dramatically induced upon the temperature downshift. Decay characteristics of the des transcript as well as the in vivo decay of B. subtilis bulk mRNA were investigated. The results showed that the stability of the des transcript as well as of bulk mRNA lasted substantially longer at 20°C than at 37°C. Functional expression of des at 37°C was achieved by exchanging its promoter with the non-cold shock spacpromoter. These data provide the first direct evidence that temperature-mediated control of transcription is the major mechanism regulating the mRNA levels of the B. subtilis desaturase. The present results also demonstrate that the only component of the desaturation system regulated by temperature is the desaturase enzyme.

scholarly journals Avaliação in vitro da adsorção de aflatoxina B1 por produtos comerciais utilizados na alimentação animal

2017 ◽  
Vol 84 (0) ◽  
Author(s):  
Raizza Eveline Escórcio Pinheiro ◽  
Carina Maricel Pereyra ◽  
Josyanne Araújo Neves ◽  
Rodrigo Maciel Calvet ◽  
Julliet Teixeira de Oliveira Santos ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Bacillus Subtilis ◽  
Bacillus Licheniformis ◽  
Lactobacillus Acidophilus ◽  
Oreochromis Niloticus ◽  
Enterococcus Faecium ◽  
Bacillus Pumilus ◽  
Bifidobacterium Bifidum

RESUMO: Objetivou-se avaliar a capacidade de adsorção in vitro de aflatoxina B1 (AFB1) por produtos comerciais utilizados na alimentação animal. Muitas pesquisas estão sendo realizadas para a descontaminação de AFB1 em alimentos. Os produtos comerciais utilizados frequentemente na alimentação de peixes, disponíveis na forma de probióticos, são formados por cepas de bactérias e leveduras utilizadas na maioria dos ensaios de adsorção de micotoxinas. Foram utilizados três produtos comerciais: A, composto por Bacillus subtilis, Bifidobacterium bifidum, Enterococcus faecium e Lactobacillus acidophilus; B, por leveduras secas de Saccharomyces cerevisiae provenientes de cervejaria; e C, por Bacillus subtilis, Bacillus licheniformis e Bacillus pumilus. Cinco suspensões da dose máxima recomendada pelo fabricante de cada produto (0; 25; 50; 75 e 100%) foram testadas contra AFB1 (1000 ng.mL-1) em microtubos para determinação da capacidade de adsorção. Para simular o pH do estômago e do intestino de tilápias do Nilo (Oreochromis niloticus) foram formuladas soluções tampão fosfato salino (PBS), com pH 1,5 e 7,5; respectivamente. Os microtubos foram introduzidos em uma centrífuga com agitação mecânica, a 37ºC por 1 h e depois centrifugados por 10 min a 14.000 rpm; os sobrenadantes foram quantificados por cromatografia líquida de alta eficiência. Os produtos comerciais, nas concentrações máximas, foram capazes de adsorver AFB1 em quantidades de 45,01 a 129,59; 123,90 a 215,59 e 209,98 a 370,73 ng.mL-1, respectivamente. Concluiu-se que todos os produtos comerciais analisados adsorvem AFB1 em condições simuladas de pH gastrointestinal e são candidatos potenciais para adsorção de AFB1 para futuros ensaios in vivo.

Antibiotic production and biocontrol activity by Bacillus subtilis CL27 and Bacillus pumilus CL45

1995 ◽  
Vol 78 (2) ◽  
pp. 97-108 ◽  
Author(s):  
C. Leifert ◽  
H. Li ◽  
Siripun Chidburee ◽  
S. Hampson ◽  
Suzanne Workman ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Pumilus ◽  
Antibiotic Production ◽  
Biocontrol Activity

Deciphering characteristics of the designer cellulosome from Bacillus subtilis WB800N via enzymatic analysis

2017 ◽  
Vol 117 ◽  
pp. 147-155 ◽  
Author(s):  
Chia-Chi Lin ◽  
Cally Joe San Yap ◽  
Shu-Chen Kan ◽  
Nai-Chi Hsueh ◽  
Liang-Yu Yang ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Enzymatic Analysis ◽  
Bacillus Subtilis Wb800n ◽  
Designer Cellulosome

Monitoring of changes in the membrane proteome during stationary phase adaptation of Bacillus subtilis using in vivo labeling techniques

PROTEOMICS ◽  
2008 ◽  
Vol 8 (10) ◽  
pp. 2062-2076 ◽  
Author(s):  
Annette Dreisbach ◽  
Andreas Otto ◽  
Dörte Becher ◽  
Elke Hammer ◽  
Alexander Teumer ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Stationary Phase ◽  
Membrane Proteome ◽  
In Vivo Labeling

scholarly journals In Bacillus subtilis, the Sirtuin Protein Deacetylase, Encoded by the srtN Gene (Formerly yhdZ), and Functions Encoded by the acuABC Genes Control the Activity of Acetyl Coenzyme A Synthetase

2009 ◽  
Vol 191 (6) ◽  
pp. 1749-1755 ◽  
Author(s):  
Jeffrey G. Gardner ◽  
Jorge C. Escalante-Semerena
Keyword(s):  
Bacillus Subtilis ◽  
Coenzyme A ◽  
Acetyl Coenzyme A ◽  
Acetyl Coenzyme ◽  
New Information ◽  
Low Concentrations ◽  
Dependent Protein ◽  
Protein Deacetylase

ABSTRACT This report provides in vivo evidence for the posttranslational control of the acetyl coenzyme A (Ac-CoA) synthetase (AcsA) enzyme of Bacillus subtilis by the acuA and acuC gene products. In addition, both in vivo and in vitro data presented support the conclusion that the yhdZ gene of B. subtilis encodes a NAD+-dependent protein deacetylase homologous to the yeast Sir2 protein (also known as sirtuin). On the basis of this new information, a change in gene nomenclature, from yhdZ to srtN (for sirtuin), is proposed to reflect the activity associated with the YdhZ protein. In vivo control of B. subtilis AcsA function required the combined activities of AcuC and SrtN. Inactivation of acuC or srtN resulted in slower growth and cell yield under low-acetate conditions than those of the wild-type strain, and the acuC srtN strain grew under low-acetate conditions as poorly as the acsA strain. Our interpretation of the latter result was that both deacetylases (AcuC and SrtN) are needed to maintain AcsA as active (i.e., deacetylated) so the cell can grow with low concentrations of acetate. Growth of an acuA acuC srtN strain on acetate was improved over that of the acuA + acuC srtN strain, indicating that the AcuA acetyltransferase enzyme modifies (i.e., inactivates) AcsA in vivo, a result consistent with previously reported in vitro evidence that AcsA is a substrate of AcuA.

scholarly journals Functional expression of complement factor I following AAV-mediated gene delivery in the retina of mice and human cells

Gene Therapy ◽  
2021 ◽  
Author(s):  
Anna K. Dreismann ◽  
Michelle E. McClements ◽  
Alun R. Barnard ◽  
Elise Orhan ◽  
Jane P. Hughes ◽  
...  
Keyword(s):  
Complement System ◽  
Functional Expression ◽  
Geographic Atrophy ◽  
Age Related Macular Degeneration ◽  
Complement Factor ◽  
Factor I ◽  
Complement Factor I ◽  
Age Related ◽  
The Complement System

AbstractDry age-related macular degeneration (AMD) is characterised by loss of central vision and currently has no approved medical treatment. Dysregulation of the complement system is thought to play an important role in disease pathology and supplementation of Complement Factor I (CFI), a key regulator of the complement system, has the potential to provide a treatment option for AMD. In this study, we demonstrate the generation of AAV constructs carrying the human CFI sequence and expression of CFI in cell lines and in the retina of C57BL/6 J mice. Four codon optimised constructs were compared to the most common human CFI sequence. All constructs expressed CFI protein; however, most codon optimised sequences resulted in significantly reduced CFI secretion compared to the non-optimised CFI sequence. In vivo expression analysis showed that CFI was predominantly expressed in the RPE and photoreceptors. Secreted protein in vitreous humour was demonstrated to be functionally active. The findings presented here have led to the formulation of an AAV-vectored gene therapy product currently being tested in a first-in-human clinical trial in subjects with geographic atrophy secondary to dry AMD (NCT03846193).

Tetramer formation of Bacillus subtilis YabJ protein that belongs to YjgF/YER057c/UK114 family

2021 ◽  
Vol 85 (2) ◽  
pp. 297-306
Author(s):  
Zui Fujimoto ◽  
Le Thi Thu Hong ◽  
Naomi Kishine ◽  
Nobuhiro Suzuki ◽  
Keitarou Kimura
Keyword(s):  
Bacillus Subtilis ◽  
Quaternary Structure ◽  
Single Amino Acid ◽  
Wild Type ◽  
Amino Acid Mutation ◽  
X Ray Crystallography ◽  
Ligand Binding Sites ◽  
Potential Ligand ◽  
Single Amino Acid Mutation

ABSTRACT Bacillus subtilis YabJ protein belongs to the highly conserved YjgF/YER057c/UK114 family, which has a homotrimeric quaternary structure. The dominant allele of yabJ gene that is caused by a single amino acid mutation of Ser103Phe enables poly-γ-glutamic acid (γPGA) production of B. subtilis under conditions where the cell-density signal transduction was disturbed by the loss of DegQ function. X-ray crystallography of recombinant proteins revealed that unlike the homotrimeric wild-type YabJ, the mutant YabJ(Ser103Phe) had a homotetrameric quaternary structure, and the structural change appeared to be triggered by an inversion of the fifth β-strand. The YabJ homotetramer has a hole that is highly accessible, penetrating through the tetramer, and 2 surface concaves as potential ligand-binding sites. Western blot analyses revealed that the conformational change was also induced in vivo by the Ser103Phe mutation.

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