A novel α-galactosidase from Fusarium oxysporum and its application in determining the structure of the gum arabic side chain

Maruta, Akiho; Yamane, Mirei; Matsubara, Midori; Suzuki, Shiho; Nakazawa, Masami; Ueda, Mitsuhiro; Sakamoto, Tatsuji

doi:10.1016/j.enzmictec.2017.04.006

Cited by 22 publications

(30 citation statements)

References 37 publications

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“…There were many reports on the cloning and expression of α‐galactosidases. Many α‐galactosidase genes from Fusarim oxysporum , Bacillus megaterium , and Rhizomucor miehei were successfully cloned and expressed . Thus, in order to have better productivity and facilitate enzyme purification, it is necessary to clone the enzyme in the near future.…”

Section: Discussionmentioning

confidence: 99%

Hydrolysis of oligosaccharides by a fungal α‐galactosidase from fruiting bodies of a wild mushroom Leucopaxillus tricolor

Geng

Fan²,

et al. 2018

J Basic Microbiol

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A novel acidic α-galactosidase (EC 3.2.1.22) designated as Leucopaxillus tricolor α-galactosidase (LTG) has been purified to homogeneity from the fruiting bodies of L. tricolor to 855-fold with a specific activity of 956 U mg by the application of chromatography and gel filtration. The molecular mass of LTG was estimated to be 60 kDa as determined by both SDS-PAGE and by gel filtration. The purified enzyme was identified by LC-MS/MS and four inner amino acid sequences were obtained. When 4-nitrophenyl α-D-glucopyranoside (pNPGal) was used as substrate, the optimal pH and optimal temperature of LTG were pH 5.0 and 50 °C, respectively. The enzyme activity was strongly inhibited by Hg , Fe , Cu , Cd , and Mn ions. The chemical modification agent N-bromosuccinimide (NBS) completely inhibited the enzyme activity of LTG, indicating the paramount importance of tryptophan residue(s) to its enzymatic activity. Besides, LTG displayed wide substrate diversity with activity toward a variety of substrates such as stachyose, raffinose, melibiose, locust bean gum, and guar gum. Given the good ability of degrading the non-digestible and flatulence-causing oligosaccharides, this fungus may become a useful source of α-galactosidase for multiple applications.

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Section: Discussionmentioning

confidence: 99%

Hydrolysis of oligosaccharides by a fungal α‐galactosidase from fruiting bodies of a wild mushroom Leucopaxillus tricolor

Geng

Fan²,

et al. 2018

J Basic Microbiol

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“…For instance, the optimum activity for F. oxysporum α-galactosidase was found at 35°C. After 1 h incubation at 40°C all the enzyme activity has been retained [23]. Optimum temperature of P. djamor α-galactosidase was reported as 53.5°C and the enzyme was preserved 68% of its initial activity after incubation at 50°C for 1 h [24].…”

Section: Stepmentioning

confidence: 96%

“…α-Galactosidases have find various industrial applications such as in su-gar industry to increase the yield of crystallized sugar, in feed industry as an animal feed additive to improve nutritional value, in pulp and paper industry to improve pulp bleaching and also in clinical applications such as for the treatment of Fabry's disease by enzyme replacement therapy and to convert B blood group to O blood group [20][21][22]. α-Galactosidases have been purified from different microbial sources with a number of purification protocols including salting-out, ion-exchange, affinity chromatography etc [23][24][25]. These protocols have generally multiple steps, need large process times and also have high operation costs.…”

Section: Introductionmentioning

confidence: 99%

Three-Phase Partitioning of α-Galactosidase from Aspergillus lentulus: Optimization of System and Characterization of Enzyme

Çamurlu

Bayraktar

Özdemir

et al. 2020

Hacettepe Journal of Biology and Chemistry

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Ü çlü-faz ayırma (TPP) tekniği α-galaktozidazın Aspergillus lentulus'dan tek adımda kısmi saflaştırılması için ilk kez başarıyla kullanıldı. Yüksek aktivite ve saflaştırma katı elde etmek için enzimin ekstraksiyon etkinliğine amonyum sülfat konsantrasyonu, ekstrakt t-butanol oranı ve pH etkisi araştırıldı. Sistemin optimum saflaştırma parametreleri %55(w/v) amonyum sülfat konsantrasyonu, 1:1 (v/v) ham ekstrakt t-butanol oranı ve pH 5.5 olarak belirlendi. Bu optimize TPP sistemi α-galaktosidaz için 5.3 saflaştırma katı ile %178 aktivite verimi oluşturdu. pH 6.5 ve 50°C'de maksimum aktivite gözlendi. α-Galaktozidaz 25-60°C sıcaklık aralığında ve pH 2.6-5.5 aralığında oldukça iyi bir kararlılık gösterdi. KM ve Vmax değerleri sırasıyla 0.365 mM ve 0.093 U olarak belirlendi. Metal iyonları ve şekerler arasında Na 2 CO 3 ve galaktoz enzim aktivitesi üzerinde kuvvetli inhibitor etkisi gösterdi. TPP ile A. lentulus'dan farklı biyokimyasal özelliklere sahip yeni bir α-galaktozidazın elde edilmesi onun çeşitli biyoteknolojik uygulamaları açısından ilgi çekici olacaktır.Anahtar Kelimeler α-galaktozidaz, biyoayırım, üçlü-faz ayırma (TPP), enzim saflaştırma, enzim karakterizasyonu.

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“…To substitute H with D, the sample (5 mg) was dissolved in D 2 O, lyophilized, and dissolved in 0.6 mL of D 2 O. 1 H-NMR spectra were recorded using a 500 MHz spectrometer (JNM-ECZ500, JEOL, Ltd., Tokyo, Japan) in D 2 O at 80°C as previously reported [19].…”

Section: Nuclear Magnetic Resonance (Nmr) Spectroscopymentioning

confidence: 99%

Identification and characterization of GH62 bacterial α-l-arabinofuranosidase from thermotolerant Streptomyces sp. SWU10 that preferentially degrades branched l-arabinofuranoses in wheat arabinoxylan

Phuengmaung

Kunishige

Sukhumsirichart

et al. 2018

Enzyme and Microbial Technology

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We previously described thermotolerant Streptomyces sp. SWU10, which produced four endo-xylanases and one xylosidase able to digest xylan backbones. To achieve arabinoxylan degradation, the swu62A gene was cloned and overexpressed in Escherichia coli, and the recombinant enzyme, termed SWUAbf62A, was characterized. The 438 amino acids of SWUAbf62A revealed Glyco_hydro_62 and closely related with putative α-l-arabinofuranosidases belonging to glycoside hydrolase family 62. SWUAbf62A was purified in two steps, Ni-affinity and size-exclusion column chromatographies, and its molecular mass without signal peptide was determined to be 49 kDa. SWUAbf62A showed optimum activity at pH 5.0 and 50 °C, and more than 70% of its initial enzymatic activity remained after incubation at pH 4.1-10.5, while SWUAbf62A lost all activity after 1 h at 60 °C. SWUAbf62A activity was stimulated by Ba, Ca, and Mn and decreased by Ag, Cu, Fe, and EDTA. SWUAbf62A had no activity towards p-nitrophenyl-α-l-arabinofuranoside or p-nitrophenyl-β-d-xylopyranoside synthetic substrates. On the other hand, SWUAbf62A had the highest activity against wheat arabinoxylan, with a specific activity of 1.29 U/mg, and was also active against sugar beet arabinan, with a specific activity of 0.14 U/mg; these results indicated that SWUAbf62A is an arabinoxylan arabinofuranohydrolase. Using H-NMR analysis, SWUAbf62A was found to release l-arabinofuranoses singly linked to O-3 of wheat arabinoxylan. In addition, SWUAbf62A acted synergistically with endo-xylanase (XynSW3) and α-l-arabinofuranosidase, which releases arabinose linked to O-3 of double-substituted xylose residues on arabinoxylan, to digest the wheat arabinoxylan. SWUAbf62A is an important debranching enzyme for hydrolysis of hemicelluloses to monosaccharides and can be applied in various industrial biotechnologies.

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A novel α-galactosidase from Fusarium oxysporum and its application in determining the structure of the gum arabic side chain

Cited by 22 publications

References 37 publications

Hydrolysis of oligosaccharides by a fungal α‐galactosidase from fruiting bodies of a wild mushroom Leucopaxillus tricolor

Hydrolysis of oligosaccharides by a fungal α‐galactosidase from fruiting bodies of a wild mushroom Leucopaxillus tricolor

Three-Phase Partitioning of α-Galactosidase from Aspergillus lentulus: Optimization of System and Characterization of Enzyme

Identification and characterization of GH62 bacterial α-l-arabinofuranosidase from thermotolerant Streptomyces sp. SWU10 that preferentially degrades branched l-arabinofuranoses in wheat arabinoxylan

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