Optimization for Production of Exo-β-1,3-glucanase (Laminarinase) from Aspergillus oryzae in Saccharomyces cerevisiae

Abstract

1) In this study, a EXGA gene code for exo-β-1,3-glucanase from Aspergillus oryzae was overexpressed and secretory produced in Saccharomyces cerevisiae. To overexpress the β-1,3-glucanase, pGInu-exgA and pAInu-exgA plasmids having GAL10 and ADH1 promoter, respectively, and exoinulinase signal sequence (Inu s.s) were constructed and introduced in S. cerevisiae SEY2102 and 2805. The recombinant β-1,3-glucanase was successfully expressed and secreted into the medium and the β-1,3-glucanase activity in 2102/pGInu-exgA and 2102/pAInu-exgA strain were 5.01 unit/mL and 4.09 unit/mL, respectively. In the 2805/pGInu-exgA and 2805/pAInu-exgA strain, the β-1,3-glucanase activity showed 3.23 unit/mL and 3.22 unit/mL, respectively. Secretory efficiency in each strain reached 95% to 98%. Subsequently, the recombinant β-1,3- Department of Biotechnology and Bioengineering, Pukyong National University, Busan 608-737, Korea glucanase was used for ethanol production. Ethanol productivity in 2102/pAInu-exgA strain was 0.83 g/L when pre-treated Laminaria japonica which has initial reducing sugar of 1.4 g/L was used as substrate. It is assumed that the polysaccharides of Laminaria japonica was effectively saccharified by recombinant β-1,3-glucanase, resulting in increase of ethanol productivity. These results suggested that recombinant β-1,3-glucanase was efficiently overexpressed and secreted in S. cerevisiae SEY2102 as host strain by using ADH1 promoter-Inu s.s system.

Construction of an expression system for the secretory production of recombinant α-agarase in yeast

“…When the EXGA gene coding for exo-b-1,3-glucanase from Aspergillus oryzae was expressed in S. cerevisiae, the glucanase activity differed only slightly between the constitutive and inducible expression systems (Kim et al 2011). However, the AgaA gene was efficiently expressed by the inducible promoter in S. cerevisiae.…”

A First Expression, Purification and Characterization of Endo-β-1,3-Glucanase from Penicillium expansum

“…β-1,3glucanases of various origins have been reported to improve the strains used in industry. For example, beta-1,3-glucanase from Aspergillus oryzae expressed in Saccharomyces cerevisiae enables yeast to degrade cellulosic materials [15,16]. Similarly, to enhance oligoglucoside hydrolysis, a thermostable β-1,3-glucanase from Trichoderma harzianum was expressed in Pichia pastoris [11,17].…”

Breeding of Ethanol-producing and Ethanol-tolerant Saccharomyces cerevisiae using Genome Shuffling

“…Following cell fusion, four candidate cells (No. 3,9,11, and 12 strains) showing thermotolerance at 40℃ were selected, and their ethanol tolerance (7% ethanol concentration) and β-1,3-glucanase activity were subsequently analyzed. All the phenotypes of the two parent cells were simultaneously expressed in one (No.…”