Purification and characterization of a novel GH1 beta-glucosidase from Jeotgalibacillus malaysiensis

et al. 2020

AMB Expr

A newly identified cellulase-producing Fusarium chlamydosporum HML278 was cultivated under solid-state fermentation of sugarcane bagasse, and two new β-glucosides enzymes (BG FH1, BG FH2) were recovered from fermentation solution by modified non-denaturing active gel electrophoresis and gel filtration chromatography. SDS-PAGE analysis showed that the molecular weight of BG FH1 and BG FH2 was 93 kDa and 52 kDa, respectively, and the enzyme activity was 5.6 U/mg and 11.5 U/mg, respectively. The optimal reaction temperature of the enzymes was 60 ℃, and the enzymes were stable with a temperature lower than 70 ℃. The optimal pH of the purified enzymes was 6.0, and the enzymes were stable between pH 4–10. Km and Vmax values were 2.76 mg/mL and 20.6 U/mg for pNPG, respectively. Thin-layer chromatography and high-performance liquid chromatography analysis showed that BG FH1and BG FH2 had hydrolysis activity toward cellobiose and could hydrolyze cellobiose into glucose. In addition, both enzymes exhibited transglycoside activity, which could use glucose to synthesize cellobiose and cellotriose, and preferentially synthesize alcohol. In conclusion, our study demonstrated that F. chlamydosporum HML278 produces heat-resistant β-glucosidases with both hydrolytic activity and transglycosidic activity, and these β-glucosidases have potential application in bioethanol and papermaking industries.

Section: The Properities Of β-Glucosidase Isolated From F Chlamydospmentioning

confidence: 99%

One-step purification of two novel thermotolerant β-1,4-glucosidases from a newly isolated strain of Fusarium chlamydosporum HML278 and their characterization

et al. 2020

AMB Expr

“…After hydrolysis of substrate into glucose, it can be further fermented to produce ethanol (Chan et al, 2016). Liew et al puri ed a new β-glucosidase BglD5 (GH1) from Jeotgalibacillus malaysiensis, and BglD5 was stable at temperature below 65℃, which promoted the cellulase hydrolysis (Liew et al, 2018 ). Kumar found that endoglucanase and βglucosidase did not lose their enzymatic activity within 60 minutes at 80 °C.…”

Section: The Hydrolysis Activity Of Bg Fh1 and Bg Fh2mentioning

confidence: 99%

One-step purification and characterization of two novel Thermotolerant β-1,4-glucosidases from a newly isolated strain of Fusarium chlamydosporum HML278

et al. 2020

Preprint

A newly screened cellulase-producing Fusarium chlamydosporum HML278 was cultivated under solid-state fermentation of sugarcane bagasse, and two new β-glucosides enzymes (BG FH1, BG FH2) from fermentation solution were recovered by modified non-denaturing active gel electrophoresis and gel filtration chromatography. SDS-PAGE analysis showed that the molecular weight of BG FH1and BG FH2 was 93 kDa and 52 kDa, respectively, and the enzyme activity was 5.6 U/mg and 11.5 U/mg, respectively. The optimal reaction temperature of the enzymes was 60 ℃, and the enzymes were stable under 70 ℃. The optimal pH of the purified enzymes was 6.0, and the enzymes were stable between pH 4–10. Km and Vmax values of 2.76 mg/mL, 20.6 U/mg for pNPG. Thin-layer chromatography and high-performance liquid chromatography analysis showed that cellobiose BG FH1and BG FH2 had hydrolysis activity and can hydrolyze cellobiose into glucose. In addition, both enzymes also exhibited transglycoside activity, which can use low molecular weight monosaccharides to synthesize cellobiose and cellotriose, and preferentially synthesize alcohol. In conclusion, our study demonstrated that F. chlamydosporum HML278 can produce heat-resistant β-glucosidase with both hydrolytic activity and transglycosidic activity, and has potential application value in bioethanol and papermaking industries.

“…After hydrolysis of substrate to glucose, which can be further fermented to produce ethanol (Chan et al, 2016). Liew et al puri ed a new β-glucosidase BglD5 (GH1) from Jeotgalibacillus malaysiensis, and BglD5 was stable at temperature below 65 ℃, which promoted the cellulase hydrolysis (Liew et al, 2018 ). Kumar found that endoglucanase and β-glucosidase retained enzymatic activity within 60 minutes at 80 °C.…”

Section: The Hydrolysis Activity Of Bg Fh1 and Bg Fh2mentioning

confidence: 99%

One-step purification of two novel thermotolerant β-1,4-glucosidases from a newly isolated strain of Fusarium chlamydosporum HML278 and their characterization

et al. 2020

Preprint

A newly identified cellulase-producing Fusarium chlamydosporum HML278 was cultivated under solid-state fermentation of sugarcane bagasse, and two new β-glucosides enzymes (BG FH1, BG FH2) were recovered from fermentation solution by modified non-denaturing active gel electrophoresis and gel filtration chromatography. SDS-PAGE analysis showed that the molecular weight of BG FH1 and BG FH2 was 93 kDa and 52 kDa, respectively, and the enzyme activity was 5.6 U/mg and 11.5 U/mg, respectively. The optimal reaction temperature of the enzymes was 60 ℃, and the enzymes were stable with a temperature lower than 70 ℃. The optimal pH of the purified enzymes was 6.0, and the enzymes were stable between pH 4-10. Km and Vmax values were 2.76 mg/mL and 20.6 U/mg for pNPG, respectively. Thin-layer chromatography and high-performance liquid chromatography analysis showed that BG FH1and BG FH2 had hydrolysis activity toward cellobiose and could hydrolyze cellobiose into glucose. In addition, both enzymes exhibited transglycoside activity, which could use glucose to synthesize cellobiose and cellotriose, and preferentially synthesize alcohol. In conclusion, our study demonstrated that F. chlamydosporum HML278 produces heat-resistant β-glucosidases with both hydrolytic activity and transglycosidic activity, and these β-glucosidases have potential application in bioethanol and papermaking industries.