Novel Chitosanase from<i>Streptomyces griseus</i>HUT 6037 with Transglycosylation Activity

Tanabe, Toshizumi; Morinaga, Kazuko; Fukamizo, Tamo; Mitsutomi, Masaru

doi:10.1271/bbb.67.354

Cited by 74 publications

(52 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…GH9 family enzymes have an (␣/␣) 6 -barrel fold, and this family of enzymes is known to contain members that exhibit different enzymatic activities, including endoglucanase (45), cellobiohydrolase (15), 1,4-␤-D-glucan glucohydrolase (43), ␤-glucosidase (40), and exo-␤-glucosaminidase (23). The GH5 family of enzymes have an (␤/␣) 8 -barrel fold, and this family also encodes diverse enzymatic activities, including endoglucanase (16), mannanase (20), mannosidase (20), xylanase (14), chitosanase (50), glucosidase (11), licheninase (25), fucosidase (61), galactanase (44), endoglycoceramidase (9), and mannan transglycosylase (6). The binding cleft in GH9 catalytic modules can hold six sugar molecules, among which four subsites (Ϫ4 to Ϫ1) are at the nonreducing end and the two remaining subsites are at the reducing end (ϩ1 to ϩ2) (45).…”

mentioning

confidence: 99%

Biochemical and Mutational Analyses of a Multidomain Cellulase/Mannanase from Caldicellulosiruptor bescii

Mackie

Cann

2012

Appl Environ Microbiol

View full text Add to dashboard Cite

Thermophilic cellulases and hemicellulases are of significant interest to the biofuel industry due to their perceived advantages over their mesophilic counterparts. We describe here biochemical and mutational analyses of Caldicellulosiruptor bescii Cel9B/ Man5A (CbCel9B/Man5A), a highly thermophilic enzyme. As one of the highly secreted proteins of C. bescii, the enzyme is likely to be critical to nutrient acquisition by the bacterium. CbCel9B/Man5A is a modular protein composed of three carbohydratebinding modules flanked at the N terminus and the C terminus by a glycoside hydrolase family 9 (GH9) module and a GH5 module, respectively. Based on truncational analysis of the polypeptide, the cellulase and mannanase activities within CbCel9B/ Man5A were assigned to the N-and C-terminal modules, respectively. CbCel9B/Man5A and its truncational mutants, in general, exhibited a pH optimum of ϳ5.5 and a temperature optimum of 85°C. However, at this temperature, thermostability was very low. After 24 h of incubation at 75°C, the wild-type protein maintained 43% activity, whereas a truncated mutant, TM1, maintained 75% activity. The catalytic efficiency with phosphoric acid swollen cellulose as a substrate for the wild-type protein was 7.2 s ؊1 ml/mg, and deleting the GH5 module led to a mutant (TM1) with a 2-fold increase in this kinetic parameter. Deletion of the GH9 module also increased the apparent k cat of the truncated mutant TM5 on several mannan-based substrates; however, a concomitant increase in the K m led to a decrease in the catalytic efficiencies on all substrates. These observations lead us to postulate that the two catalytic activities are coupled in the polypeptide.

show abstract

mentioning

confidence: 99%

Biochemical and Mutational Analyses of a Multidomain Cellulase/Mannanase from Caldicellulosiruptor bescii

Mackie

Cann

2012

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…In addition, in Cel9A C , a carbohydrate-binding module 4 (CBM4) probably contributing to binding to substrates was also found [12]. The deduced amino acid sequence consisting of 372 residues from the cel5A C gene showed 75% identity to chitosanase II from Streptomyces griseus HUT 6037, which catalyzes transglycosylation in the hydrolysis of chitooligosaccharides and exhibits CMCase activity as well [30]. Meanwhile, the deduced amino acid sequence from cel5A M (459 amino acids) was highly homologous (81% identity) to CelA from S. lividans 66 [5].…”

Section: Cloning and Nucleotide Sequencing Of Secretorymentioning

confidence: 96%

Cloning and Sequence Analysis of the Cellulase Genes Isolated from Two Cellulolytic Streptomycetes and Their Heterologous Expression in <i>Streptomyces lividans </i>

Tomotsune

Kasuga

Tsuchida

et al. 2014

Int.J.Soc.Mater.Eng.Resour.

View full text Add to dashboard Cite

Cellulosic biomass, such as wood waste and rice straw, remains unexploited despite the high content of cellulose. Streptomycetes are producers of industrially important antibiotics and most of the antibioticproducing strains are unable to utilize cellulose. We have, therefore, aimed to breed streptomycetes that produce antibiotics from cellulosic biomass. Two cellulolytic streptomycetes, i.e. Streptomyces thermocarboxydus C42 and Streptomyces argenteolus M178, were selected on the basis of carboxymethyl cellulose (CMC)-degrading activity from approximately 1,000 soil-isolated streptomycetes. Shotgun cloning experiments of cellulase genes from these genomic DNAs were performed in Streptomyces lividans as a host to find putative four cellulase genes, i.e. cel9A C and cel5A C from the C42 strain and cel5A M and cel12B M from the M178 strain. The transformants of S. lividans harboring the four genes subcloned in an expression plasmid showed secreted CMC-degrading cellulase (CMCase) activities, thus demonstrating successful cloning and heterologous expression of the cellulase genes from the cellulolytic streptomycetes. 2.2 CMC assay and Measurement of CMC-degrading cellulase (CMCase) activity Streptomyces isolates were cultivated in 2 ml of GPY and CPY liquid media at 30˚C in a test tube containing a glass bead on a reciprocal shaker at 130 strokes/min for 3 to 4 days. A portion (50 µl) of the supernatant of culture broth was placed in wells (6 mm diameter and 5 mm depth) on 1.5% (w/v) agar plates containing 0.5% (w/v) CMC sodium salt and the plates were incubated at 37˚C for 12 hours for CMC degradation [14]. Staining of the plates with 0.1% (w/v) Congo red solution and the following destaining with 1 M NaCl visualized CMC-digested clear zones formed around the wells by the diffusion of CMCase. On the other hand, the CMCase activity was measured using CMC as the substrate as described by Okada et al. [15] with some modifications. The supernatant (20 µL) of the Streptomyces culture was mixed with 80 µL of 1.25% CMC solution in 50 mM acetate buffer (pH 6.0) for incubation at 37˚C for 60 min, and the reducing sugar released was determined by the 3,5-dinitrosalicylic acid method [16]. The experiment was conducted in triplicate. One unit of CMCase activity was defined as the amount of enzyme releasing 1 µmol of glucose equivalent per minute.2.3 DNA manipulation DNA manipulations using E. coli as a host were carried out as described by Sambrook et al. [18]. DNA manipulations in Streptomyces cells were conducted as described in a manual by Kieser et al. [16]. Enzymes and kits for DNA recombination techniques were purchased from Takara Bio Inc. and Toyobo Co. Ltd., and used according to the manufacturers' instructions. Polymerase chain reaction (PCR) was conducted using an iCycler thermal cycler (Bio-Rad Laboratories Inc.) with Takara LA Taq with GC buffer (Takara Bio Inc.) and KOD-Plus-(Toyobo Co. Ltd.). All PCR primers used for this study were synthesized commercially (Hokkaido System Science Co. Ltd.) as listed in Table...

show abstract

“…Chitosanases (EC 3.2.1.132) are glycosyl hydrolases that catalyze the endohydrolysis of -1,4 linkages between GlcN residues in a partially Nacetylated chitosan (Enzyme Nomenclature, http:// www.chem.qmul.ac.uk/iubmb/enzyme/). They are produced by various bacteria [1][2][3][4][5][6][7][8][9] and fungi, [10][11][12][13][14][15] and are useful in the preparation of biofunctional chitooligosaccharides. Fukamizo et al 16) have proposed that chitosanases can be classified into three subclasses according to their specificity for the hydrolysis of the -glycosidic linkages in partially N-acetylated chitosan molecules: subclass I chitosanases [16][17][18][19][20] split both GlcN-GlcN and GlcNAc-GlcN bonds, subclass II chitosanases 3) split only the GlcN-GlcN bond, and subclass III chitosanases 5,[21][22][23] split both GlcN-GlcN and GlcN-GlcNAc bonds.…”

mentioning

confidence: 99%

Classification of Chitosanases by Hydrolytic Specificity towardN¹,N⁴-Diacetylchitohexaose

Hirano

Watanabe

Seki

et al. 2012

Bioscience, Biotechnology, and Biochemistry

Self Cite

View full text Add to dashboard Cite

Novel Chitosanase fromStreptomyces griseusHUT 6037 with Transglycosylation Activity

Cited by 74 publications

References 29 publications

Biochemical and Mutational Analyses of a Multidomain Cellulase/Mannanase from Caldicellulosiruptor bescii

Biochemical and Mutational Analyses of a Multidomain Cellulase/Mannanase from Caldicellulosiruptor bescii

Cloning and Sequence Analysis of the Cellulase Genes Isolated from Two Cellulolytic Streptomycetes and Their Heterologous Expression in <i>Streptomyces lividans </i>

Classification of Chitosanases by Hydrolytic Specificity towardN¹,N⁴-Diacetylchitohexaose

Contact Info

Product

Resources

About

Novel Chitosanase fromStreptomyces griseusHUT 6037 with Transglycosylation Activity

Cited by 74 publications

References 29 publications

Biochemical and Mutational Analyses of a Multidomain Cellulase/Mannanase from Caldicellulosiruptor bescii

Biochemical and Mutational Analyses of a Multidomain Cellulase/Mannanase from Caldicellulosiruptor bescii

Cloning and Sequence Analysis of the Cellulase Genes Isolated from Two Cellulolytic Streptomycetes and Their Heterologous Expression in <i>Streptomyces lividans </i>

Classification of Chitosanases by Hydrolytic Specificity towardN1,N4-Diacetylchitohexaose

Contact Info

Product

Resources

About

Classification of Chitosanases by Hydrolytic Specificity towardN¹,N⁴-Diacetylchitohexaose