The lipA gene of Serratia marcescens which encodes an extracellular lipase having no N-terminal signal peptide

Akatsuka, Hiroyuki; Kawai, Eri; Omori, Kenji; Komatsubara, Saburo; Shibatani, Takeji; Tosa, Tetsuya

doi:10.1128/jb.176.7.1949-1956.1994

Cited by 78 publications

(38 citation statements)

References 71 publications

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“…Another unrelated lipase, Plu2266, is similar to LipP (carboxylesterase) of Pseudomonas sp. strain B11-1 35 , and Plu2313 is highly similar to the extracellular lipase, LipA, from Serratia marcescens 36 . Plu2313 lacks an N-terminal signal peptide but contains instead, in its C terminus, four almost exact repeats of the glycine-rich motif L-x-G-G-x-G-x-D of RTX toxins, suggestive of a similar secretion pathway 37 .…”

Section: Secreted Proteinsmentioning

confidence: 99%

The genome sequence of the entomopathogenic bacterium Photorhabdus luminescens

et al. 2003

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Photorhabdus luminescens is an enterobacterium that is symbiotic with soil entomopathogenic nematodes and pathogenic to a wide range of insects. P. luminescens promotes its own transmission among susceptible insect populations using its nematode host as vector 1 . Its life cycle comprises a symbiotic stage in the nematode's gut and a virulent stage in the insect larvae, which it kills through toxemia and septicemia. After the nematode attacks a prey insect and P. luminescens is released, the bacterium produces a wide variety of virulence factors ensuring rapid insect killing. Bioconversion of the insect cadaver by exoenzymes produced by the bacteria allows the bacteria to multiply and the nematode to reproduce. During this process P. luminescens produces antibiotics to prevent invasion of the insect cadaver by bacterial or fungal competitors. Finally, elimination of competitors allows P. luminescens and the nematode to reassociate specifically before leaving the insect cadaver 2,3 .To better understand this complex life style, we determined the genome sequence of P. luminescens subspecies laumondii strain TT01 4 , a symbiont of the nematode Heterorhabditis bacteriophora isolated on Trinidad and Tobago. RESULTS General featuresStrain TT01 possesses a single circular chromosome of 5,688,987 bp with an average GC content of 42.8%. No plasmid replicon was found.A total of 4,839 protein-coding genes, including 157 pseudogenes, seven complete sets (23S, 5S and 16S) of ribosomal RNA operons and 85 tRNA genes, were predicted ( Fig. 1; Supplementary Table 1 online). Toxins against insectsMore toxin genes were predicted in the P. luminescens genome than in any other bacterial genome sequenced yet. A large number of these toxins may be involved in the killing of a wide variety of insects. Some may act synergistically or use redundancy for 'overkill' 5 , ensuring a quick death of the host. In addition, some may kill insects by interfering with their development. In the TT01 genome, two paralogs, plu4092 and plu4436, encode proteins similar to juvenile hormone esterases (JHEs) of the insect Leptinotarsa decemlineata 6 . Juvenile hormone maintains the insect in a larval state. Its inactivation by JHE allows metamorphosis to proceed. JHEs may be used to trigger the insect endocrine machinery at an inappropriate time and thus represents a promising approach for insect control 7 . These genes are located downstream of highly related orphan genes (plu4093 and plu4437), suggesting a locus duplication.The toxicity of the proteins encoded by these two loci was verified experimentally. Two Escherichia coli clones, containing the recombinant BAC1A02 and BAC8C11, were shown to be toxic toward insects. BAC1A02, which contains the locus plu4093-plu4092, exhibited substantial oral toxicity toward three mosquito species, Aedes aegypti,

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Section: Secreted Proteinsmentioning

confidence: 99%

The genome sequence of the entomopathogenic bacterium Photorhabdus luminescens

et al. 2003

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“…For lipase production, the cells of S. marcescens were cultured at 30ЊC in 500-ml flasks containing 60 ml of the lipase medium (29), which was made of 2% Meast S (Asahi Brewery Co., Ltd., Tokyo, Japan), 1% dextrin, 0.2% (NH 4 ) 2 SO 4 , 0.1% KH 2 PO 4 , 0.05% MgSO 4 ⅐ 7H 2 O, 0.01% CaCl 2 ⅐ 2H 2 O, 0.001% FeSO 4 ⅐ 7H 2 O, Tween 80 (polyoxyethylene sorbitanmonooleate), and 0.1% Colorin 102 (Sanyo Chemical Industries, Kyoto, Japan) (pH 7.0). The modified tributyrin agar plate, which was used to detect the lipase activity of transformants, was described previously (1). The concentration of ampicillin and kanamycin added was 200 g/ml (each).…”

Section: Methodsmentioning

confidence: 99%

“…The antiserum against the S. marcescens lipase used was described previously (1). Peptide Prt1 (NH 2 -HPGDYNAGEGNPTYRDVT-COOH), corresponding to amino acid residues 202 to 219 of S. marcescens metalloprotease protein, was synthesized by the t-butyloxycarbinyl synthesis strategy.…”

Section: Methodsmentioning

confidence: 99%

“…The third component of secretion machinery for the S. marcescens metalloprotease and the heme-binding protein HasA (24), which corresponds to the TolC protein for the E. coli hemolysin or the prtF EC protein for the E. chrysanthemi metalloproteases, was not identified in the has locus (23,25). The lipase and metalloprotease proteins of S. marcescens have no signal sequence at the N terminus but contain the repeated sequence in the C-terminal moiety (1). The hasDE genes involved in the metalloprotease secretion have been cloned (23).…”

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The three genes lipB, lipC, and lipD involved in the extracellular secretion of the Serratia marcescens lipase which lacks an N-terminal signal peptide

Akatsuka¹,

Kawai²,

Omori³

et al. 1995

J Bacteriol

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The extracellular lipase of Serratia marcescens Sr41, lacking a typical N-terminal signal sequence, is secreted via a signal peptide-independent pathway. The 20-kb SacI DNA fragment which allowed the extracellular lipase secretion was cloned from S. marcescens by selection of a phenotype conferring the extracellular lipase activity on the Escherichia coli cells. The subcloned 6.5-kb EcoRV fragment was revealed to contain three open reading frames which are composed of 588, 443, and 437 amino acid residues constituting an operon (lipBCD). Comparisons of the deduced amino acid sequences of the lipB, lipC, and lipD genes with those of the Erwinia chrysanthemi prtD EC , prtE EC , and prtF EC genes encoding the secretion apparatus of the E. chrysanthemi protease showed 55, 46, and 42% identity, respectively. The products of the lipB and lipC genes were 54 and 45% identical to the S. marcescens hasD and hasE gene products, respectively, which were secretory components for the S. marcescens heme-binding protein and metalloprotease. In the E. coli DH5 cells, all three lipBCD genes were essential for the extracellular secretion of both S. marcescens lipase and metalloprotease proteins, both of which lack an N-terminal signal sequence and are secreted via a signal-independent pathway. Although the function of the lipD gene seemed to be analogous to those of the prtF EC and tolC genes encoding third secretory components of ABC transporters, the E. coli TolC protein, which was functional for the S. marcescens Has system, could not replace LipD in the LipB-LipC-LipD transporter reconstituted in E. coli. These results indicated that these three proteins are components of the device which allows extracellular secretion of the extracellular proteins of S. marcescens and that their style is similar to that of the PrtDEF EC system.The 62-kDa extracellular lipase of Serratia marcescens has no typical N-terminal signal sequence, but a sequence consisting of multiple repeats of nine amino acid residues (GGXGXD XXX), which is characterized as a glycine-and aspartic acidrich region, is situated in the C-terminal moiety. This sequence was found in the following extracellular proteins of gram-negative bacteria: metalloprotease from Erwinia chrysanthemi (5, 6); hemolysin, encoded by the hlyA gene in Escherichia coli (9); leukotoxin, encoded by the lktA gene in Pasteurella haemolytica (26); cyclolysin, a multifunctional protein carrying an adenylate cyclase activity and a hemolytic activity, encoded by the cyaA gene in Bordetella pertussis (11); and Ca 2ϩ -binding protein, encoded by the nodO gene in Rhizobium leguminosarum (7). The colicin V protein, the cvaC gene product of E. coli (10), possesses a repeated glycine-rich sequence which is not homologous to the GGXGXDXXX sequence but shares some characteristics with it. Since the E. coli cells carrying the S. marcescens lipA gene encoding the lipase did not secrete the lipase protein into the medium, the lipase is expected to be secreted extracellularly via a signal peptide-independent s...

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“…marcescens has been shown for a metalloprotease [38] and a lipase [39], which are secreted by the ATP-binding cassette (ABC) protein-mediated exporter. The proteins excreted through the ABC pathway are known to contain a Gly-rich sequence that is repeated 4-36 times at the C-terminus [39,40]. However, chitinases C1 and B do not possess this type of secretion signal at their C-termini.…”

Section: Figure 7 Relationships Among Family 18 Bacterial Chitinasesmentioning

confidence: 99%

The third chitinase gene (chiC) of Serratia marcescens 2170 and the relationship of its product to other bacterial chitinases

Suzuki

Taiyoji

Sato

et al. 1999

Biochemical Journal

142

123

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The third chitinase gene (chiC) of Serratia marcescens 2170, specifying chitinases C1 and C2, was identified. Chitinase C1 lacks a signal sequence and consists of a catalytic domain belonging to glycoside hydrolase family 18, a fibronectin type III-like domain (Fn3 domain) and a C-terminal chitin-binding domain (ChBD). Chitinase C2 corresponds to the catalytic domain of C1 and is probably generated by proteolytic removal of the Fn3 and ChBDs. The loss of the C-terminal portion reduced the hydrolytic activity towards powdered chitin and regenerated chitin, but not towards colloidal chitin and glycol chitin, illustrating the importance of the ChBD for the efficient hydrolysis of crystalline chitin. Phylogenetic analysis showed that bacterial family 18 chitinases can be clustered in three subfamilies which have diverged at an early stage of bacterial chitinase evolution. Ser. marcescens chitinase C1 is found in one subfamily, whereas chitinases A and B of the same bacterium belong to another subfamily. Chitinase C1 is the only Ser. marcescens chitinase that has an Fn3 domain. The presence of multiple, divergent, chitinases in a single chitinolytic bacterium is perhaps necessary for efficient synergistic degradation of chitin.

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The lipA gene of Serratia marcescens which encodes an extracellular lipase having no N-terminal signal peptide

Cited by 78 publications

References 71 publications

The genome sequence of the entomopathogenic bacterium Photorhabdus luminescens

The genome sequence of the entomopathogenic bacterium Photorhabdus luminescens

The three genes lipB, lipC, and lipD involved in the extracellular secretion of the Serratia marcescens lipase which lacks an N-terminal signal peptide

The third chitinase gene (chiC) of Serratia marcescens 2170 and the relationship of its product to other bacterial chitinases

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