title: Construction of engineered Saccharomyces cerevisiae strain to improve that whole-cell biocatalytic production of melibiose from raffinose.
abstract: There are excessive by-products in the biocatalysis process of this whole-cell biocatalytic production of melibiose from raffinose with current Saccharomyces cerevisiae strains. To solve this problem, we constructed engineered strains based on a liquor yeast (S. cerevisiae) via gene deletion (mel1 gene), heterologous integration (fsy1 or/and ffzi1 gene from Candida magnoliae), and gene overexpression (gcr1 gene). Functional verification showed that deletion of the mel1 gene led to elimination of the reactions catalyzed by α-galactosidase, as well as elimination of the degradation of melibiose and the formation of galactose by-product. Insertion of the fsy1 or/and ffzi1 gene and overexpression of the gcr1 gene could contribute to fructose transport for enhancing the biopurification rate of the fructose by-product. Compared with the wild-type strain, the optimal engineered strain of MP8 (Δmel1::fsy1cm::ffzi1cm::gcr1sc) had improved about 30% on yield, 31% on productivity, and 36% on purity of the melibiose product.
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology of Ministry of Education, Key Laboratory of Industrial Microbiology of Tianjin, College of Bioengineering, Tianjin University of Science and Technology, No. 32, 13th. Avenue, Tianjin Economic and Development Area, Tianjin, 300457, China.|National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, No. 32, West 7th Avenue, Tianjin Airport Economic Area, Tianjin, 300308, China.
authors.forename: Yingbiao
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, No. 32, West 7th Avenue, Tianjin Airport Economic Area, Tianjin, 300308, China.
authors.forename: Yueming
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, No. 32, West 7th Avenue, Tianjin Airport Economic Area, Tianjin, 300308, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Men
authors.affiliation: Tianjin Key Laboratory on Technology Enabling Development of Clinical Therapeutics and Diagnosis, School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China.
authors.forename: Caixia
authors.initials: C
authors.lastname: Dong
authors.affiliation: National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, No. 32, West 7th Avenue, Tianjin Airport Economic Area, Tianjin, 300308, China. syx0430@hotmail.com.
authors.forename: Yuanxia
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology of Ministry of Education, Key Laboratory of Industrial Microbiology of Tianjin, College of Bioengineering, Tianjin University of Science and Technology, No. 32, 13th. Avenue, Tianjin Economic and Development Area, Tianjin, 300457, China. Zkz_12@126.com.
authors.forename: Juankun
authors.initials: J
authors.lastname: Zhang
mesh_terms: D055162:Biocatalysis; D005632:Fructose / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D017353:Gene Deletion; D007218:Industrial Microbiology; D008553:Melibiose / Q000378:metabolism*; D000074041:Microorganisms, Genetically-Modified; D015202:Protein Engineering; D011887:Raffinose / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D000519:alpha-Galactosidase / Q000378:metabolism
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D005632:Fructose; D008553:Melibiose; D000519:alpha-Galactosidase; D011887:Raffinose; D005690:Galactose
keywords: Biopurification; Engineered strain; Melibiose; Raffinose; Saccharomyces cerevisiae; Whole-cell biocatalysis