Expression, Purification, and Characterization of Recombinant α-N-Acetylgalactosaminidase Produced in the YeastPichia pastoris

Zhu, Alex; Monahan, Catherine; Wang, Zhong-Kun; Goldstein, Jack

doi:10.1006/prep.1996.0124

Cited by 31 publications

(12 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In these studies, which used a lower enzyme concentration, prolonged incubation at 37 ± C was necessary to achieve deantigenation. Another group, as reported by Goldstein (25,26). The problem with the chicken enzyme is the high concentration and low pH required for deantigenation.…”

Section: Tablementioning

confidence: 99%

Purification and Characterization of a‐N‐Acetylgalactosaminidase from Clostridium perfringens

et al. 2000

View full text Add to dashboard Cite

Summary®-N-Acetylgalactosaminidase from Clostridium perfringens is an exoglycosidase that degrades the human blood type A epitope. A highly puri ed preparation of ®-N-acetylgalactosaminidase was obtained from C. perfringens by salt precipitation, gel ltration, ionexchange chromatography, chromatofocusing, and high-pressure liquid chromatography. The nal preparation was homogeneous by sodium dodecyl sulfate -polyacrylamide gel electrophoresis, with a molecular mass of 72.1 kDa. The enzyme was highly selective for terminal N-acetyl-®-D-galactosamine residues. No other substantial glycosidase activities, speci cally neuraminidase, were detected. The pH optimum of the enzyme was between 6.5 and 7.0, and activity was unaffected by ionic strength. No protease activity was detected and enzyme activity was stable at 4 ± C for 12 months. ELISA experiments demonstrated activity against blood type A epitope. IUBMB Life, 50: 91 -97, 2000

show abstract

Section: Tablementioning

confidence: 99%

Purification and Characterization of a‐N‐Acetylgalactosaminidase from Clostridium perfringens

et al. 2000

View full text Add to dashboard Cite

show abstract

“…The specific activities of the purified rAppA Sc (658 U/mg) were lower than that of the rAppA Ec (1122 U/ mg). The decrease may be due to the difference in molecular weight caused by hyperglycosylation (Zhu et al 1996). Both purified rAppA Ec and rAppA Sc were examined and compared for its thermostability at temperature of 50-70°C, respectively.…”

Section: L E R a G G N I E L Y T Q R Y Q S S F R T L 188 841 Tggaaamentioning

confidence: 99%

Molecular Cloning of the Phytase Gene from Citrobacter braakii and its Expression in Saccharomyces cerevisiae

Kim

Lee

et al. 2006

Biotechnol Lett

View full text Add to dashboard Cite

The gene, appA, encoding phytase was cloned from a size-selected genomic library of Citrobacter braakii YH-15 by Southern hybridization using a degenerate probe based on the N-terminal amino acid sequence of the phytase. The deduced amino acid sequence of appA contained the N-terminal RHGXRXP motif and the C-terminal HD motif, which are common in histidine acid phosphatases. It also had significant homology (60% identity) with phytase from Escherichia coli, while the physical mapping analysis of appA revealed that gene organization near appA in C. braakii was similar to that in Salmonella typhimurium genome. C. braakii AppA contained five putative N-glycosylation sites. The recombinant phytases, rAppAEc and rAppASc, were produced in E. coli and Saccharomyces cerevisiae, respectively, with both being fused with C-terminal His-tag. After purification, rAppASc was shown to be hyperglycosylated by Endo-H treatment. It had greater thermostability than the wild type phytase and rAppAEc.

show abstract

“…It is difficult to convert group A RBCs because the biochemistry of A 1 antigen is more complex than that of B antigen. A variety of αNAGAs derived from human beings, chicken liver, sea squirt, yeast, niger and so on have been cloned, but the specificity and efficiency were low [8][9][10][11] . The αNAGA and endo-β-galactosidase from clostridium perfringens could convert group A 2 RBCs to O, but they could not convert group A 1 RBCs [12,13] .…”

mentioning

confidence: 99%

Human RBCs blood group conversion from A to O using a novel α-N-acetylgalactosaminidase of high specific activity

Yu¹,

Wang³

et al. 2008

Sci. Bull.

View full text Add to dashboard Cite

Expression, Purification, and Characterization of Recombinant α-N-Acetylgalactosaminidase Produced in the YeastPichia pastoris

Cited by 31 publications

References 0 publications

Purification and Characterization of a‐N‐Acetylgalactosaminidase from Clostridium perfringens

Purification and Characterization of a‐N‐Acetylgalactosaminidase from Clostridium perfringens

Molecular Cloning of the Phytase Gene from Citrobacter braakii and its Expression in Saccharomyces cerevisiae

Human RBCs blood group conversion from A to O using a novel α-N-acetylgalactosaminidase of high specific activity

Contact Info

Product

Resources

About