Preparation and Application of Trichoderma reesei Mutants with Enhanced β -Glucosidase

Mihashi²,

et al. 2019

J Wood Sci

Enzymatic biomass saccharification is an important process for bioethanol production. Hitherto, numerous cellulase cocktails (crude enzyme) have been developed to improve enzymatic activity. For this purpose, the synergy of incorporating hydrolase functionality within a cellulase cocktail is a key function. However, such synergistic action, by potentially numerous different enzyme types, on biomass tissue has not been considered despite the importance toward the realistic case of biomass saccharification. This study aims to visualize the behavior of each of the key cellulase components on biomass tissue during saccharification. Time-lapse fluorescence microscopy observations were conducted during saccharification of a thin transverse sugarcane section to monitor enzymes modified with a fluorescence dye. Statistical image analysis successfully demonstrated a unique adsorption/desorption behavior of each enzyme component. Particularly, the behavior of endoxylanase10 (Xyn10), which was recently discovered from Penicillium sp. as a high-performance xylanase, displayed remarkable adsorption on tissues of sugarcane, which accounts for the superior activity of the cellulase mixture with Xyn10.

Section: Enzyme Preparationmentioning

confidence: 99%

Selective fluorescence labeling: time-lapse enzyme visualization during sugarcane hydrolysis

Mihashi²,

et al. 2019

J Wood Sci

FEMS Microbiology Letters

“…Efficient production of/3-glucosidase required screening mutants that are derepressed versus glucose, or those that hyperproduce this enzyme. Tahoun et al [12] and Kawamori et al [13] screened hyperproducer mutants of /3-glucosidase from Trichoderma viride and Trichoderma reesei, respectively. Similar investigations were reported for other microorganisms and enzymes: glucoamylase from Aspergillus terreus [14] and endocellular/3-glucosidase from Candida wickerhamii [15].…”

Section: Introductionmentioning

confidence: 99%

Selection and study of a Candida molischiana mutant derepressed for β-glucosidase production

Janbon¹

1994

A mutant strain of Candida molischiana was selected. Analysis of the exocellular activity of Candida molischiana 35M5N grown on different carbon sources revealed that the biosynthesis of beta-glucosidase is derepressed in this yeast strain. The strain is not a hyper-producer mutant. There were no observed differences in the endocellular and parietal activities of the wild and mutant strains. However, the mutant strain produced 35-fold more enzyme than the wild-type in the culture medium with glucose as carbon source. When glucose was used as carbon source, the mutant strain produced 90% more exocellular enzyme than when cellobiose was used as the carbon source.

Secretion of Trichoderma reesei β-glucosidase by Saccharomyces cerevisiae

Cummings¹,

Fowler²

1996

Current Genetics

An intronless form of the bgl1 gene encoding an extracellular beta-glucosidase from Trichoderma reesei was expressed in the yeast Saccharomyces cerevisiae under the control of the yeast GAL1 promoter. Transformation of a yeast strain with this vector resulted in transformants that produce and secrete active beta-glucosidase into the growth medium. Additionally, active recombinant beta-glucosidase protein was shown to be localized predominantly in the periplasmic space by using a p-nitrophenyl beta-D-glycoside hydrolysis assay against fractionated yeast cells. The apparent size of the recombinant enzyme was 10-15 kDa larger than that of the native form. Treatment of the recombinant beta-glucosidase with endoglycosidase-H indicated the apparent increase in size was due to N-linked glycosylation.