Swedish
Albanian
Arabic
Armenian
Azerbaijani
Belarusian
Bengali
Bosnian
Catalan
Czech
Danish
Deutsch
Dutch
English
Estonian
Finnish
Français
Greek
Haitian Creole
Hebrew
Hindi
Hungarian
Icelandic
Indonesian
Irish
Italian
Japanese
Korean
Latvian
Lithuanian
Macedonian
Mongolian
Norwegian
Persian
Polish
Portuguese
Romanian
Russian
Serbian
Slovak
Slovenian
Spanish
Swahili
Swedish
Turkish
Ukrainian
Vietnamese
Български
中文(简体)
中文(繁體)
Wei sheng wu xue bao = Acta microbiologica Sinica 2012-May

[A novel alpha-galactosidase from Arthrobacter sp. GN14 isolated from Grus nigricollis feces: gene cloning, heterologous expression and characterization].

Endast registrerade användare kan översätta artiklar
Logga in Bli medlem
Länken sparas på Urklipp
Junpei Zhou
Lu Pan
Junjun Li
Xianghua Tang
Zunxi Huang

Nyckelord

Abstrakt

OBJECTIVE

Cloning and heterologously expressing the alpha-galactosidase gene (agaAGN14) from Arthrobacter sp. GN14 isolated from feces of black-neck crane (Grus nigricollis).

METHODS

The full-length agaAGN14 was cloned based on degenerate PCR and GC TAIL-PCR (thermal asymmetric interlaced PCR), ligated into pET-28a (+) vector and expressed in Escherichia coli BL21 (DE3) cells. The recombinant alpha-galactosidase (rAgaAGN14) was purified to electrophoretic homogeneity by Ni(2+)-NTA metal chelating affinity chromatography, and then the enzyme characterizations were determined. Amino acids sequences of agaAGN14 (AgaAGN14) and alpha-galactosidases from Actinobacteria and gastrointestinal microorganisms were aligned and used for constructing a neighbor-joining phylogenetic tree.

RESULTS

The 2109-bp full-length agaAGN14 (66.8% GC content) encodes a 702-residue polypeptide (AgaAGN14; 77.5 kDa). AgaAGN14 showed the highest identity of 53.7% with alpha-galactosidases in public databases, and < 43% identities with alpha-galactosidases from gastrointestinal microorganisms. AgaAGN14 was put in a phylogenetic branch sharing the catalytic motifs KWD and SDXXDXXXR, and close to alpha-galactosidases from soil microorganisms and far from alpha-galactosidases from gastrointestinal microorganisms. The purified rAgaAGN14 efficiently hydrolyzed pNPG, raffinose, melibiose, stachyose, rapeseed meal and cottonseed meal; showed apparent optimal at pH 6.0 and 45 degrees C, stability and activity (> 50%) at pH 6.0-9.0, and activities of 28%, 30% and 80% at 10 degrees C, 20 degrees C and 37 degrees C, respectively; exhibited K(m), V(max) and k(cat) values of 0.41 mmol/L, 18.28 micromol/min/mg and 25.36 s(-1), respectively, using pNPG as the substrate at 45 degrees C and pH 6.5; strongly inhibited by Ag+, Hg2+ and SDS, partial inhibited by K+, Ca2+, Mn2+, Fe3+, Ni2+, Cu2+ and beta-mercaptoethanol, and little influenced by Co2+, Pb2+, Zn2+, Mg2+, Na+ and EDTA.

CONCLUSIONS

The Arthrobacter strain isolated from feces of Grus nigricollis, and the sequence analysis, phylogenetic analysis, heterologous expression and recombinant enzyme's biochemical characterizations of an alpha-galactosidase from Arthrobacter strain were first reported. rAgaAGN14 was a novel alpha-galactosidase.

Gå med på vår
facebook-sida

Den mest kompletta databasen med medicinska örter som stöds av vetenskapen

  • Fungerar på 55 språk
  • Växtbaserade botemedel som stöds av vetenskap
  • Örter igenkänning av bild
  • Interaktiv GPS-karta - märka örter på plats (kommer snart)
  • Läs vetenskapliga publikationer relaterade till din sökning
  • Sök efter medicinska örter efter deras effekter
  • Organisera dina intressen och håll dig uppdaterad med nyheterna, kliniska prövningar och patent

Skriv ett symptom eller en sjukdom och läs om örter som kan hjälpa, skriv en ört och se sjukdomar och symtom den används mot.
* All information baseras på publicerad vetenskaplig forskning

Google Play badgeApp Store badge