Efficiency Analysis and Mechanism Insight of that Whole-Cell Biocatalytic Production of Melibiose from Raffinose with Saccharomyces cerevisiae

被引:26
作者
Zhou, Yingbiao [1 ,2 ]
Zhu, Yueming [2 ]
Dai, Longhai [2 ]
Men, Yan [2 ]
Wu, Jinhai [3 ]
Zhang, Juankun [1 ]
Sun, Yuanxia [2 ]
机构
[1] Tianjin Univ Sci & Technol, Key Lab Ind Fermentat Microbiol, Minist Educ, Coll Biotechnol, Tianjin 300457, Peoples R China
[2] Chinese Acad Sci, Natl Engn Lab Ind Enzymes, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China
[3] Shanxi Normal Univ, Sch Food Sci, Linfen 041000, Peoples R China
基金
中国国家自然科学基金; 国家高技术研究发展计划(863计划);
关键词
Whole-cell biocatalysis; Saccharomyces cerevisiae; Characteristics analysis; Melibiose; Raffinose; SIGNAL-SEQUENCE TRAP; ALPHA-GALACTOSIDASE; FRUCTOSE/H+ SYMPORTER; SECRETED PROTEINS; YEAST-STRAIN; INVERTASE; GENE; FSY1; CLONING;
D O I
10.1007/s12010-016-2220-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Melibiose is widely used as a functional carbohydrate. Whole-cell biocatalytic production of melibiose from raffinose could reduce its cost. However, characteristics of strains for whole-cell biocatalysis and mechanism of such process are unclear. We compared three different Saccharomyces cerevisiae strains (liquor, wine, and baker's yeasts) in terms of concentration variations of substrate (raffinose), target product (melibiose), and by-products (fructose and galactose) in whole-cell biocatalysis process. Distinct difference was observed in whole-cell catalytic efficiency among three strains. Furthermore, activities of key enzymes (invertase, alpha-galactosidase, and fructose transporter) involved in process and expression levels of their coding genes (suc2, mel1, and fsy1) were investigated. Conservation of key genes in S. cerevisiae strains was also evaluated. Results show that whole-cell catalytic efficiency of S. cerevisiae in the raffinose substrate was closely related to activity of key enzymes and expression of their coding genes. Finally, we summarized characteristics of producing strain that offered advantages, as well as contributions of key genes to excellent strains. Furthermore, we presented a dynamic mechanism model to achieve some mechanism insight for this whole-cell biocatalytic process. This pioneering study should contribute to improvement of whole-cell biocatalytic production of melibiose from raffinose.
引用
收藏
页码:407 / 423
页数:17
相关论文
共 22 条
[1]   Three-dimensional Structure of Saccharomyces Invertase ROLE OF A NON-CATALYTIC DOMAIN IN OLIGOMERIZATION AND SUBSTRATE SPECIFICITY [J].
Angela Sainz-Polo, M. ;
Ramirez-Escudero, Mercedes ;
Lafraya, Alvaro ;
Gonzalez, Beatriz ;
Marin-Navarro, Julia ;
Polaina, Julio ;
Sanz-Aparicio, Julia .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2013, 288 (14) :9755-9766
[2]   Crystallization and preliminary X-ray diffraction analysis of the invertase from Saccharomyces cerevisiae [J].
Angela Sainz-Polo, M. ;
Lafraya, Alvaro ;
Polo, Aitana ;
Marin-Navarro, Julia ;
Polaina, Julio ;
Sanz-Aparicio, Julia .
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 2012, 68 :1538-1541
[3]   Fsy1, the sole hexose-proton transporter characterized in Saccharomyces yeasts, exhibits a variable fructose:H+ stoichiometry [J].
Anjos, Jorge ;
de Sousa, Helena Rodrigues ;
Roca, Christophe ;
Cassio, Fernanda ;
Luttik, Marijke ;
Pronk, Jack T. ;
Salema-Oom, Madalena ;
Goncalves, Paula .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 2013, 1828 (02) :201-207
[4]   An Efficient Genome-Wide Fusion Partner Screening System for Secretion of Recombinant Proteins in Yeast [J].
Bae, Jung-Hoon ;
Sung, Bong Hyun ;
Kim, Hyun-Jin ;
Park, Soon-Ho ;
Lim, Kwang-Mook ;
Kim, Mi-Jin ;
Lee, Cho-Ryong ;
Sohn, Jung-Hoon .
SCIENTIFIC REPORTS, 2015, 5
[5]   2 DIFFERENTIALLY REGULATED MESSENGER-RNAS WITH DIFFERENT 5' ENDS ENCODE SECRETED AND INTRACELLULAR FORMS OF YEAST INVERTASE [J].
CARLSON, M ;
BOTSTEIN, D .
CELL, 1982, 28 (01) :145-154
[6]   FSY1, a horizontally transferred gene in the Saccharomyces cerevisiae EC1118 wine yeast strain, encodes a high-affinity fructose/H+ symporter [J].
Galeote, Virginie ;
Novo, Maite ;
Salema-Oom, Madalena ;
Brion, Christian ;
Valerio, Elisabete ;
Goncalves, Paula ;
Dequin, Sylvie .
MICROBIOLOGY-SGM, 2010, 156 :3754-3761
[7]   Signal-sequence trap in mammalian and yeast cells: A comparison [J].
Galliciotti, G ;
Schneider, H ;
Wyder, L ;
Vitaliti, A ;
Wittmer, M ;
Ajmo, M ;
Klemenz, R .
JOURNAL OF MEMBRANE BIOLOGY, 2001, 183 (03) :175-182
[8]   The repressor Rgt1 and the cAMP-dependent protein kinases control the expression of the SUC2 gene in Saccharomyces cerevisiae [J].
Gancedo, Juana M. ;
Flores, Carmen-Lisset ;
Gancedo, Carlos .
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 2015, 1850 (07) :1362-1367
[9]   FSY1, a novel gene encoding a specific fructose/H+ symporter in the type strain of Saccharomyces carlsbergensis [J].
Gonçalves, P ;
de Sousa, HR ;
Spencer-Martins, I .
JOURNAL OF BACTERIOLOGY, 2000, 182 (19) :5628-5630
[10]   Stability of rituximab in freeze-dried formulations containing trehalose or melibiose under different relative humidity atmospheres [J].
Heljo, Ville P. ;
Filipe, Vasco ;
Romeijn, Stefan ;
Jiskoot, Wim ;
Juppo, Anne M. .
JOURNAL OF PHARMACEUTICAL SCIENCES, 2013, 102 (02) :401-414