scholarly journals A novel amidase signature family amidase from the marine actinomyceteSalinispora arenicolaCNS-205

2018 ◽  
Author(s):  
Ma Yanling ◽  
Zhang Xinfeng ◽  
Zeng Rong
Keyword(s):  
Bioinformatics Analysis ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Acyl Transferase ◽  
Optimal Temperature ◽  
Aryl Acylamidase ◽  
Marine Actinomycete ◽  
Potential Applications ◽  
New Gene ◽  
Substrate Spectrum

AbstractWe cloned a new gene from the amidase signature (AS) family, designatedam, from the marine actinomyceteSalinispora arenicolaCNS-205. As indicated by bioinformatics analysis and site-directed mutagenesis, the AM protein belonged to the AS family. AM was expressed, purified, and characterised inEscherichia coliBL21 (DE3), and the AM molecular mass was determined to be 51 kDa. The optimal temperature and pH were 40 °C and pH 8.0, respectively. AM exhibited a wide substrate spectrum and showed amidase, aryl acylamidase, and acyl transferase activities. AM had high activity towards aromatic and aliphatic amides. The AM substrate specificity for anilides was very narrow; only propanil could be used as an effective substrate. The extensive substrate range of AM indicates it may have broad potential applications in biosynthetic processes and biodegradation.

scholarly journals A Novel Amidase Signature Family Amidase from the Marine Actinomycete Salinispora arenicola CNS-205

2019 ◽  
Vol 13 (4) ◽  
pp. 557-563 ◽  
Author(s):  
Yanling Ma ◽  
Xinfeng Zhang ◽  
Tianmei Chen ◽  
Fulai Liu ◽  
Shuting Fang ◽  
...  
Keyword(s):  
Bioinformatics Analysis ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Optimal Temperature ◽  
Gene Encoding ◽  
Aryl Acylamidase ◽  
Marine Actinomycete ◽  
Potential Applications ◽  
New Gene ◽  
Substrate Spectrum

We cloned a new gene from the amidase signature (AS) family, designated am , from the marine actinomycete Salinispora arenicola CNS-205. As indicated by bioinformatics analysis and site-directed mutagenesis, the AM (a gene encoding a putative amidase) protein belonged to the AS family. AM was expressed, purified, and characterised in Escherichia coli BL21 (DE3), and the AM molecular mass was determined to be 51 kDa. The optimal temperature and pH were 40 °C and pH 8.0, respectively. AM exhibited a wide substrate spectrum and showed amidase, aryl acylamidase, and acyl transferase activities. AM had high activity towards aromatic and aliphatic amides. The AM substrate specificity for anilides was very narrow, and only propanil could be used as an effective substrate. The extensive substrate range of AM indicates it may have broad potential applications in biosynthetic processes and biodegradation.

scholarly journals A new twist of rubredoxin function in M. tuberculosis

2020 ◽  
Author(s):  
Tatsiana Sushko ◽  
Anton Kavaleuski ◽  
Irina Grabovec ◽  
Anna Kavaleuskaya ◽  
Daniil Vakhrameev ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Isothermal Calorimetry ◽  
Cytochrome P450s ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Biological Processes ◽  
Single Charge ◽  
Iron Starvation ◽  
Potential Applications ◽  
Electron Carriers

AbstractElectron transfer mediated by metalloproteins drives many biological processes. Rubredoxins are ubiquitous iron-containing electron carriers that play important roles in bacterial adaptation to changing environmental conditions. In Mycobacterium tuberculosis, oxidative and acidic stresses as well as iron starvation induce rubredoxin expression. However, their functions during M. tuberculosis infection is unknown. In the present work, we show that rubredoxin B (RubB) supports catalytic activity of mycobacterial cytochrome P450s, CYP124, CYP125, and CYP142, which are important for bacterial viability and pathogenicity. We solved the crystal structure of RubB and characterized the interaction between RubB and CYPs using site-directed mutagenesis. Mutations that neutralized single charge on the surface of RubB did not dramatically decrease activity of studied CYPs, and isothermal calorimetry (ITC) experiments indicated that interactions are transient and not highly specific. Our findings suggest that a switch from ferredoxins to rubredoxins support CYP activity in M. tuberculosis-infected macrophages. Our electrochemical experiments suggest potential applications of RubB in biotechnology.

A peek in the micro-sized world: a review of design principles, engineering tools, and applications of engineered microbial community

2020 ◽  
Vol 48 (2) ◽  
pp. 399-409
Author(s):  
Baizhen Gao ◽  
Rushant Sabnis ◽  
Tommaso Costantini ◽  
Robert Jinkerson ◽  
Qing Sun
Keyword(s):  
Microbial Community ◽  
Microbial Communities ◽  
Environmental Remediation ◽  
Real Life ◽  
Design Principles ◽  
Microbial Interactions ◽  
Potential Applications ◽  
New Gene ◽  
Global Biogeochemical Cycles ◽  
Synthetic Microbial Communities

Microbial communities drive diverse processes that impact nearly everything on this planet, from global biogeochemical cycles to human health. Harnessing the power of these microorganisms could provide solutions to many of the challenges that face society. However, naturally occurring microbial communities are not optimized for anthropogenic use. An emerging area of research is focusing on engineering synthetic microbial communities to carry out predefined functions. Microbial community engineers are applying design principles like top-down and bottom-up approaches to create synthetic microbial communities having a myriad of real-life applications in health care, disease prevention, and environmental remediation. Multiple genetic engineering tools and delivery approaches can be used to ‘knock-in' new gene functions into microbial communities. A systematic study of the microbial interactions, community assembling principles, and engineering tools are necessary for us to understand the microbial community and to better utilize them. Continued analysis and effort are required to further the current and potential applications of synthetic microbial communities.

The site-directed mutagenesis and prokaryotic expression of midkine gene in Cynoglossus semilaevis

2013 ◽  
Vol 37 (3) ◽  
pp. 330
Author(s):  
Yanan WANG ◽  
Xudong LIU ◽  
Linlin MU ◽  
Zhipeng LIU ◽  
Chunmei LI ◽  
...  
Keyword(s):  
Prokaryotic Expression ◽  
Cynoglossus Semilaevis ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis
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