Isolation and biochemical characterization of an endo-1,3-β-glucanase from Streptomyces sioyaensis containing a C-terminal family 6 carbohydrate-binding module that binds to 1,3-β-glucan
The GenBank accession number for the sequence reported in this paper is AF21741.
A gene encoding 1,3-β-glucanase was isolated from Streptomyces sioyaensis based on an activity plate assay. Analysis of the deduced amino acid sequence of the gene revealed that the matured 1,3-β-glucanase has two functional domains separated by a stretch of nine glycine residues. The N-terminal domain shares sequence similarity with bacterial endo-1,3-β-glucanases classified in glycosyl hydrolase family 16 (GHF 16), while the C-terminal domain is a putative carbohydrate-binding module (CBM) grouped into CBM family 6. To characterize the function of each domain, both the full-length and the CBM-truncated versions of the protein were expressed in Escherichia coli and purified to homogeneity. Biochemical data suggest that the glycosyl hydrolase domain preferentially catalyses the hydrolysis of glucans with 1,3-β linkage, and has an endolytic mode of action. Binding assay indicated that the C-terminal CBM binds to various insoluble β-glucans (1,3-, 1,3–1,4- and 1,4- linkages) but not to xylan, a primary binding target for most members of CBM family 6. The full-length and the CBM-truncated proteins had similar specific activity (units per mol of hydrolase domain) on soluble 1,3-β-glucan, whereas the former had much stronger specific activity on insoluble 1,3-β-glucans, suggesting that the C-terminal CBM enhances the activity of the S. sioyaensis 1,3-β-glucanase against insoluble substrates, presumably by increasing the frequency of encounter events between the hydrolase domain and the substrate.
AltschulS. F.,
MaddenT. L.,
SchäfferA.,
ZhangA. J. Z.,
MillerW.,
LipmanD. J.
1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402[CrossRef]
BradfordM. M.
1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254[CrossRef]
BucciagliaP. A.,
SmithA. G.
1994; Cloning and characterization of Tag 1, a tobacco anther β-1,3-glucanase expressed during tetrad dissolution. Plant Mol Biol 24:903–914[CrossRef]
CastresanaC.,
de CarvalhoF.,
GheysenG.,
HabetsM.,
InzeD.,
van MontaguM.
1990; Tissue-specific and pathogen-induced regulation of a Nicotiana plumbaginifolia β-1,3-glucanase gene. Plant Cell 2:1131–1143
ChangM. M.,
CulleyD. E.,
HadwigerL. A.
1993; Nucleotide sequence of a pea ( Pisum sativum L.) β-1,3-glucanase gene. Plant Physiol 101:1121–1122[CrossRef]
ChenC. H.,
HuangJ. W.,
TzengD. S.
2000; Development of PMS 502- Streptomyces biopesticide and evaluation of its efficacy on the control of crop fungal diseases. Plant Pathol Bull 9:193 in Chinese
de la CruzJ.,
Pintor-ToroJ. A.,
BenitezT.,
LlobellA.,
RomeroL. C.
1995; A novel endo-β-1,3-glucanase, BGN13.1, involved in the mycoparasitism of Trichoderma harzianum
. J Bacteriol 177:6937–6945
de LooseM.,
AlliotteT.,
GheysenG.,
GenetelloC.,
GielenJ.,
SoetaertP,
van MontaguM.,
InzeD.
1988; Primary structure of a hormonally regulated β-glucanase of Nicotiana plumbaginifolia
. Gene 70:13–23[CrossRef]
FerrerP.,
HalkierT.,
HedegaardL.,
SavvaD.,
DiersI.,
AsenjoJ. A.
1996; Nucleotide sequence of a β-1,3-glucanase isoenzyme IIA gene of Oerskovia xanthineolytica LL G109 ( Cellulomonas cellulans ) and initial characterization of the recombinant enzyme expressed in Bacillus subtilis
. J Bacteriol 178:4751–4757
FontesC. M. G. A.,
ClarkeJ. H.,
HazlewoodG. P.,
FernandesT. H.,
GilbertH. J.,
FerreiraL. M. A.
1998; Identification of tandemly repeated type VI cellulose-binding domains in an endoglucanase from the aerobic soil bacterium Cellvibrio mixtus. Appl Microbiol Biotechnol 49:552–559[CrossRef]
GueguenY.,
VoorhorstW. G. B.,
van der OostJ.,
de VosW. M.
1997; Molecular and biochemical characterization of an endo-β-1,3-glucanase of the hyperthermophilic archaeon Pyrococcus furiosus
. J Biol Chem 272:31258–31264[CrossRef]
HahnM.,
KeitelT.,
HeinemannU.
1995; Crystal and molecular structure at 0·16-nm resolution of the hybrid Bacillus endo-1,3–1,4-β-d-glucan 4-glucanohydrolase H (A16-M). Eur J Biochem 232:849–858[CrossRef]
HenrissatB.,
BairochA.
1993; New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 293:781–788
HudspethR. L.,
HobbsS. L.,
AndersonD. M.,
GrulaJ. W.
1996; Characterization and expression of chitinase and 1,3-β-glucanase genes in cotton. Plant Mol Biol 31:911–916[CrossRef]
JuncosaM.,
PonsJ.,
DotT.,
QuerolE.,
PlanasA.
1994; Identification of active site carboxylic residues in Bacillus licheniformis 1,3–1,4-β-d-glucan 4-glucanohydrolase by site-directed mutagenesis. J Biol Chem 269:14530–14535
KeitelT.,
SimonO.,
BorrissR.,
HeinemannU.
1993; Molecular and active-site structure of a Bacillus 1,3–1,4-β-glucanase. Proc Natl Acad Sci USA 90:5287–5291[CrossRef]
KrahM.,
MisselwitzR.,
PolitzO.,
TomsenK. K.,
WelfleH.,
BorrissR.
1998; The laminarinase from thermophilic eubacterium Rhodothermus marinus : conformation, stability, and identification of active site carboxylic residues by site directed mutagenesis. Eur J Biochem 257:101–111[CrossRef]
Rios-HernandezM.,
Dos-SantosN.,
Silvia-CardosoJ.,
Bello-GarcigaJ. L.,
PedrosoM.
1994; Immunopharmacological studies of β-1,3-glucan. Arch Med Res 25:179–180
RyanE. M.,
WardO. P.
1985; Study of the effect of β-1,3-glucanase from basidiomycete QM 806 on yeast extract production. Biotechnol Lett 7:409–412[CrossRef]
ShenS. H.,
ChretienP.,
BastienL.,
SlilatyS. N.
1991; Primary sequence of the glucanase gene from Oerskovia xanthineolytica : expression and purification of the enzyme from Escherichia coli
. J Biol Chem 15:1058–1063
SpilliaertR.,
HreggvidssonG. O.,
KristjanssonJ. K.,
EggertssonG.,
PalsdottirA.
1994; Cloning and sequencing of a Rhodothermus marinus gene, bglA , coding for a thermostable β-glucanase and its expression in Escherichia coli
. Eur J Biochem 224:923–930[CrossRef]
SunJ. L.,
SakkaK.,
KaritaS.,
KimuraT.,
OhmiyaK.
1998; Adsorption of Clostridium stercorarium xylanase A to insoluble xylan and the important of the CBDs to xylan hydrolysis. J Ferment Bioeng 85:63–68[CrossRef]
SunL.,
GurnonJ. R.,
AdamsB. J.,
GravesM. V.,
van EttenJ. L.
2000; Characterization of a β-1,3-glucanase encoded by chlorella virus PBCV-1. Virology 276:27–36[CrossRef]
TommeP.,
CreaghA. L.,
KilburnD. G.,
HaynesC. A.
1996; Interaction of polysaccharides with the N-terminal cellulose-binding domain of Cellulomonas fimi CenC. I. Binding specificity and calorimetric analysis. Biochemistry 35:13885–13894[CrossRef]
WatanabeT.,
KasaharaN.,
AidaK.,
TanakaH.
1992; Three N-terminal domains of β-1,3-glucanase A1 are involved in binding to insoluble β-1,3-glucan. J Bacteriol 174:186–190
YamamotoM.,
EzureT.,
WatanabeT.,
TanakaH.,
AonoR.
1998; C-terminal domain of β-1,3-glucanase H in Bacillus circulans IAM1165 has a role in binding to insoluble β-1,3-glucan. FEBS Lett 433:41–43[CrossRef]
YiS. Y.,
HwangB. K.
1997; Purification and antifungal activity of a basic 34 kDa β-1,3-glucanase from soybean hypocotyls inoculated with Phytophthora sojae f. sp. glycines. Mol Cells 7:408–413
ZverlovV. V.,
VolkovI. Y.,
VelikodvorskayaT. V.,
SchwarzW. H.
1997; Highly thermostable endo-1,3-β-glucanase (laminarinase) LamA from Thermotoga neapolitana : nucleotide sequence of the gene and characterization of the recombinant gene product. Microbiology 143:1701–1708[CrossRef]
ZverlovV. V.,
VolkovI. Y.,
VelikodvorskayaG. A.,
SchwarzW. H.
2001; The binding pattern of two carbohydrate-binding modules of laminarinase Lam16A from Thermotoga neapolitana : differences in β-glucan binding within family CBM4. Microbiology 147:621–629
Isolation and biochemical characterization of an endo-1,3-β-glucanase from Streptomyces sioyaensis containing a C-terminal family 6 carbohydrate-binding module that binds to 1,3-β-glucanThe GenBank accession number for the sequence reported in this paper is AF21741.