Purification and characterization of thermostable α-galactosidase from Ganoderma lucidum
α-Galactosidase was purified from a fresh fruiting body of Ganoderma lucidum by precipitation with ammonium sulfate and column chromatographies with DEAE-Sephadex and Con A-Sepharose. The purified enzyme was homogeneous on polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was si...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
2014
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| Online Access: | http://www.scopus.com/inward/record.url?eid=2-s2.0-2442419823&partnerID=40&md5=2d81fd7bbae1df8033661e67f49e9282 http://cmuir.cmu.ac.th/handle/6653943832/6040 |
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| Institution: | Chiang Mai University |
| Language: | English |
| Summary: | α-Galactosidase was purified from a fresh fruiting body of Ganoderma lucidum by precipitation with ammonium sulfate and column chromatographies with DEAE-Sephadex and Con A-Sepharose. The purified enzyme was homogeneous on polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was similar to that of Mortierella vinacea α-galactosidase. The molecular mass of the enzyme was about 56 kDa by SDS-polyacrylamide gel electrophoresis, and about 249 kDa by gel filtration column chromatography. The optimum pH and temperature were 6.0 and 70°C, respectively. The enzyme was fully stable to heating at 70°C for 30 min. It hydrolyzed p-nitrophenyl-α-D- galactopyranoside (Km = 0.4 mM) but hydrolyzed little o-nitrophenyl-α-D-galactopyranoside. It also hydrolyzed melibiose, raffinose, and stachyose. The enzyme catalyzed the transgalactosylation reaction which synthesized melibiose. The product was confirmed by various analyses. |
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