Characterization of β-N-acetylglucosaminidase from a marine Pseudoalteromonas sp. for application in N-acetyl-glucosamine production

Preparative Biochemistry & Biotechnology
Ha Ju ParkDockyu Kim

Abstract

The psychrotolerant Pseudoalteromonas issachenkonii PAMC 22718 was isolated for its high exo-acting chitinase activity in the Kara Sea, Arctic. An exo-acting chitinase (W-Chi22718) was homogeneously purified from the culture supernatant of PAMC 22718, the molecular weight of which was estimated to be approximately 112 kDa. Due to its β-N-acetylglucosaminidase activity, W-Chi22718 was able to produce N-acetyl-D-glucosamine (GlcNAc) monomers from chitin oligosaccharide substrates. W-Chi22718 displayed chitinase activity from 0 to 37°C (optimal temperature of 30°C) and maintained activity from pH 6.0 to 9.0 (optimal pH of 7.6). W-Chi22718 exhibited a relative activity of 13 and 35% of maximal activity at 0 and 10°C, respectively, which is comparable to the activities of previously characterized, cold-adapted bacterial chitinases. W-Chi22718 activity was enhanced by K+, Ca2+, and Fe2+, but completely inhibited by Cu2+ and SDS. We found that W-Chi22718 can produce much more (GlcNAcs) from colloidal chitin, working together with previously characterized cold-active endochitinase W-Chi21702. Genome sequencing revealed that the corresponding gene (chi22718_IV) was 2,856 bp encoding a 951 amino acid protein with a calculated molecular w...Continue Reading

References

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Feb 12, 2002·International Journal of Systematic and Evolutionary Microbiology·Elena P IvanovaValery V Mikhailov
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Jun 12, 2013·Applied Microbiology and Biotechnology·Long LiuJian Chen

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Citations

Nov 17, 2017·Applied Microbiology and Biotechnology·Rui ZhangZunxi Huang

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Datasets Mentioned

BETA
GQ885144
PAMC
22718

Methods Mentioned

BETA
protein assay
gel filtration
size exclusion chromatography
ion-exchange

Software Mentioned

CD
BLASTP
EzTaxon server
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