title: Mating type and ploidy effect on the β-glucosidase activity and ethanol-producing performance of Saccharomyces cerevisiae with multiple δ-integrated bgl1 gene.
abstract: In order to investigate the effect of mating type and ploidy on enzymatic activity and fermentation performance in yeast with multiple δ-integrated foreign genes, eight ploidy series strains were constructed. The initial haploid strain BGL-a was shown to contain about 19 copies of the bgl1 gene. In rich media containing 2% (w/v) sugar the specific activities of BGL-aα were lower than those of BGL-aa or BGL-αα, which indicates the existence of mating type effects. While the maximum OD660 decreased with rising ploidy, the biomass yield showed no significant difference between the eight strains and the specific activities (expressed as U/mL or U/mg DCW) showed little to no variation. When cellobiose was used as the carbon source and β-glucosidase substrate, β-glucosidase was expressed more quickly and at higher levels than in glucose-containing media. The maximum specific activitiy values obtained were 19.07U/mL and 19.39U/mL for BGL-αα and BGL-aa, repsectively. The anaerobic biomass and ethanol-producing performance in rich media containing 10% cellobiose showed no significant difference among the eight strains. Their maximal ethanol concentrations and corresponding yields ranged from 40.27 to 43.46g/L and 77.56 to 83.71%, respectively. When the acid- and alkali-pretreated corncob (10% solids content) was used, the diploid BGL-aα fermented the best. When urea was used as the only supplemented nutrient, the ethanol titer and yield were 35.65g/L and 83.69%, respectively, while a control experiment using industrial Angel yeast with exogenous β-glucosidase addition gave values of 37.93g/L and 89.04%. The combined effects of δ-integration of bgl1, ploidy and mating type result in BGL-aa or BGL-αα being the optimal choice for enzyme production and BGL-aα being more suitable for cellulosic ethanol fermentation. These results provide valuable information for future yeast breeding and utilization efforts.
authors.affiliation: Tianjin R&D Center for Petrochemical Technology, Tianjin University, Tianjin 300072, China; School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China. Electronic address: wjj2013@tju.edu.cn.
authors.forename: Jianjun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Tianjin R&D Center for Petrochemical Technology, Tianjin University, Tianjin 300072, China. Electronic address: myy@tju.edu.cn.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Tianjin R&D Center for Petrochemical Technology, Tianjin University, Tianjin 300072, China. Electronic address: zhangkun@tju.edu.cn.
authors.forename: Kun
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Tianjin R&D Center for Petrochemical Technology, Tianjin University, Tianjin 300072, China; School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China. Electronic address: huajunyang@163.com.
authors.forename: Huajun
authors.initials: H
authors.lastname: Yang
authors.affiliation: Tianjin R&D Center for Petrochemical Technology, Tianjin University, Tianjin 300072, China. Electronic address: liuc@tju.edu.cn.
authors.forename: Cheng
authors.initials: C
authors.lastname: Liu
authors.affiliation: Tianjin R&D Center for Petrochemical Technology, Tianjin University, Tianjin 300072, China; School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China. Electronic address: slzhou@tju.edu.cn.
authors.forename: Shaolan
authors.initials: S
authors.lastname: Zou
authors.affiliation: Tianjin R&D Center for Petrochemical Technology, Tianjin University, Tianjin 300072, China. Electronic address: hjf@tju.edu.cn.
authors.forename: Jiefang
authors.initials: J
authors.lastname: Hong
authors.affiliation: Tianjin R&D Center for Petrochemical Technology, Tianjin University, Tianjin 300072, China; Key Laboratory for Green Chemical Technology of Ministry of Education, Tianjin University, Tianjin 300072, China. Electronic address: mhzhang@tju.edu.cn.
authors.forename: Minhua
authors.initials: M
authors.lastname: Zhang
mesh_terms: D057005:Bioengineering; D018533:Biomass; D000431:Ethanol / Q000032:analysis / Q000378:metabolism*; D005285:Fermentation; D049770:Genes, Mating Type, Fungal / Q000235:genetics*; D043326:Glucan 1,3-beta-Glucosidase / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D001617:beta-Glucosidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D000431:Ethanol; D001617:beta-Glucosidase; C468487:EXG1 protein, S cerevisiae; D043326:Glucan 1,3-beta-Glucosidase
keywords: Ethanol; Mating-type; Ploidy; β-Glucosidase; δ-Integration