Cloning and nucleotide sequence of the α-galactosidase cDNA from Cyamopsis tetragonoloba (guar)

Overbeeke, Nico; Aj, Fellinger; My, Toonen; D, van Wassenaar; Ct, Verrips

doi:10.1007/bf00027314

Cited by 73 publications

(39 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Substrate specificity has been scarcely investigated on (X-galactosyl stubs because it is difficult to get such kinds of oligosaccharides. We found that galactomannooligosaccharides having (X-galactosyl stubs could be prepared from a hydrolyzate of copra galactomannan with fJ-mannanase 2 ) and also found (X-galactosidase from Aspergillus niger 5-16 acted on the (X-galactosyl stubs, but the enzyme from A. niger [5][6][7][8][9][10][11][12][13][14][15][16] has not yet been purified. This paper deals with the purification of (X-galactosidase from P. purpurogenum No.…”

Section: -mentioning

confidence: 99%

Purification and Some Properties ofα-Galactosidase fromPenicillium purpurogenum

Shibuya

Kobayashi

Park

et al. 1995

Bioscience, Biotechnology, and Biochemistry

View full text Add to dashboard Cite

Section: -mentioning

confidence: 99%

Purification and Some Properties ofα-Galactosidase fromPenicillium purpurogenum

Shibuya

Kobayashi

Park

et al. 1995

Bioscience, Biotechnology, and Biochemistry

View full text Add to dashboard Cite

“…The guar -galactosidase has the unique and commercially attractive property of being able to release galactose from galactomannan, resulting in a galactomannan with improved gelling properties (Critchley 1987). The gene encoding guar -galactosidase has been cloned (Overbeeke et al 1989) and to study the enzyme in more detail in prokaryotes it was expressed and secreted by B. subtilis DB104 using the B. amyloliquefaciens -amylase signal peptide. Although this protease-deficient strain efficiently sectreted the -galactosidase in the exponential growth phase, residual protease activity caused a considerable decrease in the enzyme activity upon prolonged cultivation (Overbeeke et al 1990).…”

Section: Introductionmentioning

confidence: 99%

Production of secreted guar α-galactosidase by Lactococcus lactis

Leenhouts

Bolhuis

Ledeboer

et al. 1995

Applied Microbiology and Biotechnology

View full text Add to dashboard Cite

Production of secreted guar α-galactosidase by Lactococcus lactis Leenhouts, K.J.; Bolhuis, A.; Ledeboer, A.; Venema, G.; Kok, J. Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Abstract A plant -galactosidase gene was inserted in the expression vector pGKV259. The resulting plasmid pGAL2 consisted of the replication functions of the broad-host-range lactococcal plasmid pWV01, the lactococcal promoter P59, and the DNA sequences encoding the -amylase signal sequence from Bacillus amyloliquefaciens and the mature part of the -galactosidase from Cyamopsis tetragonoloba (guar).¸ac-tococcus cells of strain MG1363 harbouring this vector produced the plant -galactosidase and secreted the enzyme efficiently as judged by Western blotting and activity assays. Expression levels of up to 4.3 mg extracellular -galactosidase g (dry weight) of biomass\ were achieved in standard laboratory batch cultures. The -galactosidase produced by¸actococcus was active on the chromogenic substrate 5-bromo-4-chloro-3-indolyl -D-galactopyranoside, the trisaccharide raffinose and on the galactomannan substrate, guar gum.

show abstract

“…Genes encoding ␣-Gals have been cloned from various sources, including humans (3), plants (20,33), yeasts (12), filamentous fungi (4,19,26,28), and bacteria (1,2,13,17,18). ␣-Gals from eukaryotes show a significant degree of similarity and are grouped into family 27.…”

mentioning

confidence: 99%

Purification and Characterization of the RecombinantThermussp. Strain T2 α-Galactosidase Expressed inEscherichia coli

Ishiguro

Kaneko

Kuno

et al. 2001

Appl Environ Microbiol

View full text Add to dashboard Cite

The nucleotide sequence of the Thermus sp. strain T2 DNA coding for a thermostable ␣-galactosidase was determined. The deduced amino acid sequence of the enzyme predicts a polypeptide of 474 amino acids (M r , 53,514). The observed homology between the deduced amino acid sequences of the enzyme and ␣-galactosidase from Thermus brockianus was over 70%. Thermus sp. strain T2 ␣-galactosidase was expressed in its active form in Escherichia coli and purified. Native polyacrylamide gel electrophoresis and gel filtration chromatography data suggest that the enzyme is octameric. The enzyme was most active at 75°C for p-nitrophenyl-␣-Dgalactopyranoside hydrolysis, and it retained 50% of its initial activity after 1 h of incubation at 70°C. The enzyme was extremely stable over a broad range of pH (pH 6 to 13) after treatment at 40°C for 1 h. The enzyme acted on the terminal ␣-galactosyl residue, not on the side chain residue, of the galactomanno-oligosaccharides as well as those of yeasts and Mortierella vinacea ␣-galactosidase I. The enzyme has only one Cys residue in the molecule. para-Chloromercuribenzoic acid completely inhibited the enzyme but did not affect the mutant enzyme which contained Ala instead of Cys, indicating that this Cys residue is not responsible for its catalytic function.␣-Galactosidases (␣-Gals) are known to occur widely in microorganisms, plants, and animals, and some of them have been purified and characterized (5). ␣-Gals catalyze the hydrolysis of 1,6-linked ␣-galactose residues from oligosaccharides and polymeric galactomannans (19,27,28). In the sugar beet industry, ␣-Gals have been used to increase the sucrose yield by eliminating raffinose, which prevents the crystallization of beet sugar (31). Raffinose and stachyose in beans are known to cause flatulence. ␣-Gal has the potential to alleviate these symptoms, for instance, in the treatment of soybean milk (6).We have studied the substrate specificity of ␣-Gals from eukaryotes by using galactomanno-oligosaccharides, such as 6 3 -mono-␣-D-galactopyranosyl-␤-1,4-mannotriose (Gal 3 Man 3 ) and 63 -mono-␣-D-galactopyranosyl-␤-1,4-mannotetraose (Gal 3 Man 4 ). The structures of these galactomanno-oligosaccharides are shown in Fig. 1. Mortierella vinacea ␣-Gal I (11) and yeast ␣-Gals (32) are specific for Gal 3 Man 3, having an ␣-galactosyl residue (designated the terminal ␣-galactosyl residue) attached to the O-to-6 position of the nonreducing end mannose of ␤-1,4-mannotriose. On the other hand, Aspergillus niger 5-16 ␣-Gal (12) and Penicillium purpurogenum ␣-Gal (27) show a preference for Gal 3 Man 4 , having an ␣-galactosyl residue (designated as the side chain ␣-galactosyl residue) attached to the O-to-6 position of the third mannose from the reducing end of ␤-1,4-mannotetraose. The M. vinacea ␣-Gal II (28) acts on both substrates to almost equal extents. These facts indicate that eukaryotic ␣-Gals were classified into three groups based on the substrate specificity of these galactomanno-oligosaccharides.Genes encoding ␣-Gals have been cloned ...

show abstract

Cloning and nucleotide sequence of the α-galactosidase cDNA from Cyamopsis tetragonoloba (guar)

Cited by 73 publications

References 30 publications

Purification and Some Properties ofα-Galactosidase fromPenicillium purpurogenum

Purification and Some Properties ofα-Galactosidase fromPenicillium purpurogenum

Production of secreted guar α-galactosidase by Lactococcus lactis

Purification and Characterization of the RecombinantThermussp. Strain T2 α-Galactosidase Expressed inEscherichia coli

Contact Info

Product

Resources

About