Mutagenic effects on the fluorescence of tryptophan residues in bacteriophage T4 lysozyme: correlation with dynamics

1992 ◽  
Author(s):  
Bruce S. Hudson ◽  
Dan Harris
Keyword(s):  
Bacteriophage T4 ◽  
T4 Lysozyme ◽  
Tryptophan Residues ◽  
Mutagenic Effects

scholarly journals Purification of Bacteriophage T4 Lysozyme

1968 ◽  
Vol 243 (2) ◽  
pp. 391-397 ◽  
Author(s):  
Akira Tsugita ◽  
Masayori Inouye ◽  
Eric Terzaghi ◽  
George Streisinger
Keyword(s):  
Bacteriophage T4 ◽  
T4 Lysozyme

scholarly journals Evaluation of Feasibility of Using the Bacteriophage T4 Lysozyme to Improve the Hydrolysis and Biochemical Methane Potential of Secondary Sludge

Energies ◽  
2019 ◽  
Vol 12 (19) ◽  
pp. 3644
Author(s):  
Sangmin Kim ◽  
Seung-Gyun Woo ◽  
Joonyeob Lee ◽  
Dae-Hee Lee ◽  
Seokhwan Hwang
Keyword(s):  
Cell Wall ◽  
Bacterial Cell ◽  
Wastewater Treatment Plants ◽  
Biogas Production ◽  
Bacteriophage T4 ◽  
Suspended Solid ◽  
T4 Lysozyme ◽  
Biochemical Methane Potential ◽  
Secondary Sludge ◽  
Methane Potential

Anaerobic digestion (AD) of secondary sludge is a rate-limiting step due to the bacterial cell wall. In this study, experiments were performed to characterize secondary sludges from three wastewater treatment plants (WWTPs), and to investigate the feasibility of using bacteriophage lysozymes to speed up AD by accelerating the degradation of bacterial cell walls. Protein was the main organic material (67.7% of volatile solids in the sludge). The bacteriophage T4 lysozyme (T4L) was tested for hydrolysis and biochemical methane potential. Variations in the volatile suspended solid (VSS) concentration and biogas production were monitored. The VSS reduction efficiencies by hydrolysis using T4L for 72 h increased and ranged from 17.8% to 26.4%. Biogas production using T4L treated sludges increased and biogas production was increased by as much as 82.4%. Biogas production rate also increased, and the average reaction rate coefficient of first-order kinetics was 0.56 ± 0.02/d, which was up to 47.5% higher compared to the untreated samples at the maximum. Alphaproteobacteria, Betaproteobacteria, Flavobacteriia, Gammaproteobacteria, and Sphingobacteriia were major microbial classes in all sludges. The interpretation of the microbial community structure indicated that T4L treatment is likely to increase the rate of cell wall digestion.

scholarly journals Genetically Encoded Biosensor-Based Screening for Directed Bacteriophage T4 Lysozyme Evolution

2020 ◽  
Vol 21 (22) ◽  
pp. 8668
Author(s):  
Seung-Gyun Woo ◽  
Seong Keun Kim ◽  
Baek-Rock Oh ◽  
Seung-Goo Lee ◽  
Dae-Hee Lee
Keyword(s):  
High Throughput Screening ◽  
Bacteriophage T4 ◽  
T4 Lysozyme ◽  
Screening System ◽  
Random Mutation ◽  
Secondary Sludge ◽  
Enzyme Screening ◽  
Model Protein ◽  
Genetically Encoded Biosensor ◽  
Lysozyme Evolution

Lysozyme is widely used as a model protein in studies of structure–function relationships. Recently, lysozyme has gained attention for use in accelerating the degradation of secondary sludge, which mainly consists of bacteria. However, a high-throughput screening system for lysozyme engineering has not been reported. Here, we present a lysozyme screening system using a genetically encoded biosensor. We first cloned bacteriophage T4 lysozyme (T4L) into a plasmid under control of the araBAD promoter. The plasmid was expressed in Escherichia coli with no toxic effects on growth. Next, we observed that increased soluble T4L expression decreased the fluorescence produced by the genetic enzyme screening system. To investigate T4L evolution based on this finding, we generated a T4L random mutation library, which was screened using the genetic enzyme screening system. Finally, we identified two T4L variants showing 1.4-fold enhanced lytic activity compared to native T4L. To our knowledge, this is the first report describing the use of a genetically encoded biosensor to investigate bacteriophage T4L evolution. Our approach can be used to investigate the evolution of other lysozymes, which will expand the applications of lysozyme.

Reexamination of the role of Asp20 in catalysis by bacteriophage T4 lysozyme

Biochemistry ◽  
1991 ◽  
Vol 30 (39) ◽  
pp. 9457-9463 ◽  
Author(s):  
Larry W. Hardy ◽  
Anthony R. Poteete
Keyword(s):  
Bacteriophage T4 ◽  
T4 Lysozyme
Sign in / Sign up

Export Citation Format

Share Document