Heterologous expression of human granzyme K in Bacillus subtilis and characterization of its hydrolytic activity in vitro

1998 ◽  
Vol 27 (2) ◽  
pp. 117-124 ◽  
Author(s):  
Lilia M. Bab ◽  
Sienna Yoast ◽  
Mark Dreyer ◽  
Brian F. Schmidt
2018 ◽  
Vol 657 ◽  
pp. 78-88 ◽  
Author(s):  
Chinmayi R. Kaundinya ◽  
Handanahal S. Savithri ◽  
K. Krishnamurthy Rao ◽  
Petety V. Balaji

2018 ◽  
Vol 200 (17) ◽  
Author(s):  
Olga Ramaniuk ◽  
Martin Převorovský ◽  
Jiří Pospíšil ◽  
Dragana Vítovská ◽  
Olga Kofroňová ◽  
...  

ABSTRACTThe σIsigma factor fromBacillus subtilisis a σ factor associated with RNA polymerase (RNAP) that was previously implicated in adaptation of the cell to elevated temperature. Here, we provide a comprehensive characterization of this transcriptional regulator. By transcriptome sequencing (RNA-seq) of wild-type (wt) and σI-null strains at 37°C and 52°C, we identified ∼130 genes affected by the absence of σI. Further analysis revealed that the majority of these genes were affected indirectly by σI. The σIregulon, i.e., the genes directly regulated by σI, consists of 16 genes, of which eight (thedhbandykuoperons) are involved in iron metabolism. The involvement of σIin iron metabolism was confirmed phenotypically. Next, we set up anin vitrotranscription system and defined and experimentally validated the promoter sequence logo that, in addition to −35 and −10 regions, also contains extended −35 and −10 motifs. Thus, σI-dependent promoters are relatively information rich in comparison with most other promoters. In summary, this study supplies information about the least-explored σ factor from the industrially important model organismB. subtilis.IMPORTANCEIn bacteria, σ factors are essential for transcription initiation. Knowledge about their regulons (i.e., genes transcribed from promoters dependent on these σ factors) is the key for understanding how bacteria cope with the changing environment and could be instrumental for biotechnologically motivated rewiring of gene expression. Here, we characterize the σIregulon from the industrially important model Gram-positive bacteriumBacillus subtilis. We reveal that σIaffects expression of ∼130 genes, of which 16 are directly regulated by σI, including genes encoding proteins involved in iron homeostasis. Detailed analysis of promoter elements then identifies unique sequences important for σI-dependent transcription. This study thus provides a comprehensive view on this underexplored component of theB. subtilistranscription machinery.


2018 ◽  
Vol 120 ◽  
pp. 657-664 ◽  
Author(s):  
Xin Cui ◽  
Yuechen Jiang ◽  
Liuyi Chang ◽  
Lei Meng ◽  
Junhong Yu ◽  
...  

2020 ◽  
Vol 147 ◽  
pp. 890-897 ◽  
Author(s):  
Zhikuan Wang ◽  
Xiaoguang Li ◽  
Jiewei Tian ◽  
Yiwen Chu ◽  
Yongqiang Tian

2010 ◽  
Vol 192 (11) ◽  
pp. 2900-2907 ◽  
Author(s):  
Maarten Groeneveld ◽  
Ruud G. J. Detert Oude Weme ◽  
Ria H. Duurkens ◽  
Dirk Jan Slotboom

ABSTRACT Bacterial secondary transporters of the DctA family mediate ion-coupled uptake of C4-dicarboxylates. Here, we have expressed the DctA homologue from Bacillus subtilis in the Gram-positive bacterium Lactococcus lactis. Transport of dicarboxylates in vitro in isolated membrane vesicles was assayed. We determined the substrate specificity, the type of cotransported ions, the electrogenic nature of transport, and the pH and temperature dependence patterns. DctA was found to catalyze proton-coupled symport of the four C4-dicarboxylates from the Krebs cycle (succinate, fumurate, malate, and oxaloacetate) but not of other mono- and dicarboxylates. Because (i) succinate-proton symport was electrogenic (stimulated by an internal negative membrane potential) and (ii) the divalent anionic form of succinate was recognized by DctA, at least three protons must be cotransported with succinate. The results were interpreted in the light of the crystal structure of the homologous aspartate transporter GltPh from Pyrococcus horikoshii.


2016 ◽  
Vol 43 (6) ◽  
pp. 729-740 ◽  
Author(s):  
Tuyishime Philibert ◽  
Zhiming Rao ◽  
Taowei Yang ◽  
Junping Zhou ◽  
Genshu Huang ◽  
...  

2008 ◽  
Vol 3 (1) ◽  
pp. 26-31 ◽  
Author(s):  
Yu Qiao ◽  
Xiaobing Chen ◽  
Hongbiao Ding ◽  
Ming Yue

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