Biochemical Analysis of a Novel Lipolytic Enzyme YvdO fromBacillus subtilis168

Kato, Shiro; Yoshimura, Teizo; Hemmi, Hisashi; Moriyama, Ryuichi

doi:10.1271/bbb.90704

Cited by 6 publications

(5 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…PLP (AFC70055.1; Rahman et al, 2010 ), Bacillus subtilis . YvdO (QJR48018.1; Kato et al, 2010 ). The conserved catalytic residues Ser214 and Asp407 are indicated at the amino acid position by a star.…”

Section: Resultsmentioning

confidence: 99%

Rv3091, An Extracellular Patatin-Like Phospholipase in Mycobacterium tuberculosis, Prolongs Intracellular Survival of Recombinant Mycolicibacterium smegmatis by Mediating Phagosomal Escape

et al. 2020

View full text Add to dashboard Cite

Patatin-like phospholipases (PLPs) are important virulence factors of many pathogens. However, there are no prevailing studies regarding PLPs as a virulence factor of Mycobacterium tuberculosis (Mtb). Analysis of Rv3091, a putative protein of Mtb, shows that it belongs to the PLPs family. Here, we cloned and expressed the rv3091 gene in Mycobacterium smegmatis and, subsequently, conducted protein purification and characterization. We show that it possesses phospholipase A 1 , phospholipase A 2 , and lipase activity. We confirm the putative active site residues, namely, Ser214 and Asp407, using site directed mutagenesis. The Rv3091 is an extracellular protein that alters the colony morphology of M. smegmatis. The presence of Rv3091 enhances the intracellular survival capability of M. smegmatis in murine peritoneal macrophages. Additionally, it promotes M. smegmatis phagosomal escape from macrophages. Moreover, Rv3091 significantly increased the survival of M. smegmatis and aggravated lesions in C57BL/6 J murine lungs in vivo. Taken together, our results indicate that Rv3091 as an extracellular PLP that is critical to the pathogenicity of mycobacterium as it allows mycobacterium to utilize phospholipids for its growth and provides resistance to phagosome killing, resulting in its enhanced intracellular survival.

show abstract

Section: Resultsmentioning

confidence: 99%

Rv3091, An Extracellular Patatin-Like Phospholipase in Mycobacterium tuberculosis, Prolongs Intracellular Survival of Recombinant Mycolicibacterium smegmatis by Mediating Phagosomal Escape

et al. 2020

View full text Add to dashboard Cite

show abstract

“…On the other hand, the enzymes from bacteria have been less characterized. ExoU of P. aeruginosa was characterized as a PLA 2 enzyme [Sato et al, 2005], while protein YvdO of Bacillus subtilis appears to have lysophospholipase activity [Kato et al, 2010]. For pathogenic bacteria, patatin-like proteins have been shown to serve as virulence factors for the invasion of host cells, as in the case of ExoU and its homologous proteins in other species [Hauser et al, 1998].…”

Section: Discussionmentioning

confidence: 99%

“…DOI: 10.1159/000491698 beyond a few hours [Fu et al, 2015;Kato et al, 2010;Wei et al, 2015;Wemheuer et al, 2013]. PLP_2.9 also showed acyltransferase activity since the addition of methanol to the enzymatic reaction mixture containing PC resulted in the formation of fatty acid methyl esters.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a Novel Thermostable Enzyme from Thermus sp. 2.9 with Phospholipase and Acyltransferase Activities

et al. 2018

View full text Add to dashboard Cite

Phospholipases are classified in different enzyme families according to the ester bond they cleave within phospholipids. The use of phospholipases in industrial processes has prompted the search for new enzymes with differential properties. A gene encoding a novel phospholipase (PLP_2.9) was identified in the genome of the thermophilic strain Thermus sp. 2.9. The analysis of the primary sequence unveiled a patatin-like domain. The alignment of the amino acid sequence of PLP_2.9 to other bacterial patatin-related proteins showed that the four blocks characteristic of this type of phospholipases and the amino acids representing the catalytic dyad are conserved in this protein. PLP_2.9 was overexpressed in Escherichia coli and the purified enzyme was characterized biochemically. PLP_2.9 hydrolyzed p-nitrophenyl palmitate at alkaline pH over a wide range of temperatures (55–80°C), showing high thermostability. PLP_2.9 displayed phospholipase A and acyltransferase activities on egg yolk phosphatidylcholine. Due to its high thermostability, PLP_2.9 has potential applications as a catalyst in several industrial processes.

show abstract

“…Another group of bacterial patatin-like proteins are much smaller and consist mainly of the patatin-like domain. One example is the lipolytic enzyme YvdO, present in dormant spores of Bacillus subtilis, and which can hydrolyse p-nitrophenyl esters of short and medium chain length (C2-C10) as well as the lysophosphatidylcholine 1-myristoyl-2-lyso-sn-glycero-3phosphocholine (Kato et al, 2010). Other examples are the patatin-like proteins SMc00930 and SMc01003 from S. meliloti.…”

Section: Patatin-like Lipases In Bacteriamentioning

confidence: 99%

“…For spectrophotometric measurements, we used an enzymatic assay similar to the one described for the PLP Yvdo from B. subtilis (Kato et al, 2010). Our standard assay mixture (1.0 ml) contained 625 μM p-NP ester, 50 mM Tris-HCl buffer (pH 8.5), 150 mM NaCl, 0.2% Triton X-100, 2.5% dimethylsulfoxide and the enzyme (1-100 μg protein of cellfree extracts).…”

Section: Enzymatic Assaysmentioning

confidence: 99%

Fatty acid‐releasing activities in Sinorhizobium meliloti include unusual diacylglycerol lipase

Sahonero‐Canavesi

Sohlenkamp

Sandoval-Calderón

et al. 2015

Environmental Microbiology

View full text Add to dashboard Cite

Summary Phospholipids are well known for their membrane forming properties and thereby delimit any cell from the exterior world. In addition, membrane phospholipids can act as precursors for signals and other biomolecules during their turnover. Little is known about phospholipid signalling, turnover and remodelling in bacteria. Recently, we showed that a FadD-deficient mutant of Sinorhizobium meliloti, unable to convert free fatty acids to their coenzyme A derivatives, accumulates free fatty acids during the stationary phase of growth. Enzymatic activities responsible for the generation of these free fatty acids were unknown in rhizobia. Searching the genome of S. meliloti, we identified a potential lysophospholipase (SMc04041) and two predicted patatin-like phospholipases A (SMc00930, SMc01003). Although SMc00930 as well as SMc01003 contribute to the release of free fatty acids in S. meliloti, neither one can use phospholipids as substrates. Here we show that SMc01003 converts diacylglycerol to monoacylglycerol and a fatty acid, and that monoacylglycerol can be further degraded by SMc01003 to another fatty acid and glycerol. A SMc01003-deficient mutant of S. meliloti transiently accumulates diacylglycerol, suggesting that SMc01003 also acts as diacylglycerol lipase (DglA) in its native background. Expression of the DglA lipase in Escherichia coli causes lysis of cells in stationary phase of growth.

show abstract

Biochemical Analysis of a Novel Lipolytic Enzyme YvdO fromBacillus subtilis168

Cited by 6 publications

References 39 publications

Rv3091, An Extracellular Patatin-Like Phospholipase in Mycobacterium tuberculosis, Prolongs Intracellular Survival of Recombinant Mycolicibacterium smegmatis by Mediating Phagosomal Escape

Rv3091, An Extracellular Patatin-Like Phospholipase in Mycobacterium tuberculosis, Prolongs Intracellular Survival of Recombinant Mycolicibacterium smegmatis by Mediating Phagosomal Escape

Characterization of a Novel Thermostable Enzyme from <b><i>Thermus</i></b> sp. 2.9 with Phospholipase and Acyltransferase Activities

Fatty acid‐releasing activities in Sinorhizobium meliloti include unusual diacylglycerol lipase

Contact Info

Product

Resources

About