Saccharomyces cerevisiae 由来グリセロール脱水素
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1. http www caa go jp foods index4 html BOEHZEUUIDE S8 1 H Onishi 1963 van Zyl and Prior 1990 Onishi 1963 Iwaki et al 2001 Watanabe et al 20042. C GCYl YPR1 S cerevisiae 2 6XHis GCY1 YPR1 BGI1805 ORF Ce XCHiS HA Protein A ZZ N C GCY1 YPR1 Bio Rad 36 E XL WE HA A280nm 1A 1B 1c iD UV2 C UVI 1A 1B 1C 1D a LI Li 1 1 3 13 Ir 9 Ut 21 T 1 8 I dE 164 1j i 13 A 12 I kt ea daai Doa a as i a aia 1 os 2038 0 8 0 7 0 6 0 5 0 4 03 02 01 4 f 1 1 1 i i i b H i H E i 1 im E i F i ERRORES GoM MOWENSORCOR EURO BOUM TET
3. ij om VC Saccharomyces cerevisiae Kluyvermyces lactis S cerevisiae N C Cheng H Jiang N Shen A and Feng Y 2005 Molecular cloning and functional expression of D arabitol dehydroge nase gene from Gluconobacter oxydans in Escherichia coli FE
4. GCY1 38 E XL WE HA 5 Cellulomonas sp G3512 Enterobacter aerogenes G4783 A Cellulomonas sp G3512 B Enterbacter aerogenes G4783 100 YPR1 3 GST YPR1 pEGH ORF N S GST 6 xHis YPRI GST GST YPRI
5. Ki N C 1 S cerevisiae GCY1 TPA7 ORF BG1805 His pEGH S BOAT M ORF 2 S S Powder ura l Difco 25 1s 75 4g Ile 0 45 g Val 2 25g Ade 0 3g Arg 0 3g His 0 3g Leu 0 45g Lys 0 45g Met 0 3g Phe 0 75g Trp 0 3g Tyr 0 45g
6. RETE EDER ZN Y GST YPR1 2 4 B 5 5 A Cellulomonas sp S7 G3512 5 B Enterobactor aerogenes G4783 Oliveira et al 2006 Dominguez et al 2010
7. 1 C YPRI GCYI 2 GCY1 Jv SDS PAGE CCY7 35KDa E 6 XHis 19KDa T 54KDa 1D ERO HA GCY1 YPR1 R3 1D M 1A 1B 1C 1D 200 150 100 75 B 50 2 CCY7 Profinia SDS M 1A IB 1C 1D Profinia Saccharomyces cerevisiae o7 Ka kDa M 1A 1B 1C 1D M 1A 1B 1C 1D 250 250 150 150 100 100 75 75
8. polyhydroxy alkane Jw CEU JutreJubT c Der wee S qe D D D LO B ETAT TF RENT oA Onishi 1963 van Zyl and Prior 1990 BESC D Bk 4 D ET HEETE 2011 3 25 2011 7 22
9. 2 SC ura SC ura 500ml S Powder ura 3 75 p 10g 1 1 SC ura 500ml S Powder ura 3 75g bg 5g 69602 7 3 XYP 500ml Difco 2 15g Becton Dickinson 30 g 30g 3 2 9 SC ura CCY7 YPRI 30 C 3 EE 2 D def pee EH 090 Wu e SC ura 3ml CIEN L 30C 3 H fe EL 1ml 2 SC ura 100ml 30C CARE dE L 7z OD600nm 6 3 XYP 50ml
10. 8 4 ml 1 7ml 500 mM JB x amp pe B 18 pH10 0 6ml 1M 0 lmD IM 0 3mD 10mM NAD 0 1mD 0 1ml U 1800 340nm 20 1 S cerevisiae GCY1 YPR1 S cerevisiae GCYI YPRI Costenoble et al 2000 S cerevisiae ORF BG 1805
11. Minamihara and Suzuki 1997 Kluyveromyces lactis D HE 3 7z S cerevisiae Dujon et al 2004 Gelperin et al 2009 S cerevisiae GCYI YPRI http www yeastgenome org 6 T
12. Gelperin et aL 2009 BG 1805 GALI KEIT iA T A25 Gelperin et al 2009 OCY7 YPRI GCCY7 GCYl YPRI YPR1 pH rU pH 9 5 Cheng et ab 2005 Hallborn et al 1995 Lin 1961 Link et al 2005 Stein et al 1997 Wong et al 1993 GCY1 YPR1
13. Mem Fac Agr Ehime Univ 33 56 33 39 2011 Saccharomyces cerevisiae KH Hong mei Zuao Yudai Sumaisur Hiroki TaNAKA and Yasuo WATANABE Purification of Glycerol Dehydrogenases from Saccharomyces cerevisiae and the Substrate Specificity of Bacterial Enzymes Abstract We have obtained the dehydrogenases being specific for mannitol and arabitol with the gene cloning and over expression in yeast cells in order to measure the content of various sugar alcohols by using the dehydrogenases In this study we carried out the isolations of glycerol dehydrogenases from Saccharomyces cerevisiae and determined its substrate specificities Target proteins were purified by affinity chromatography with the addition of tag at N terminus or C terminus by gene recombination The obtained enzymes showed the low specificity for sugar alcohols However we found the bacterial enzymes that showed the high specificity for glycerol in commercial products These glycerol dehydrogenases could be available in the enzymatic analysis of glycerol content
14. 24 1 1 9 SC ura 3 ml 30C 3 lml 100 ml 30 C C 3 RE OD600nm 2 4 1500 g 10 4 1500Xg 10 4 C 5ml 300mM KC 50mM pH8 2 15m Saccharomyces cerevisiae WRZ Y U v Bi7K 3 amp BE 38 OO T8 38 L AA EHE SS OO EUER IE 39 Z 15g Vibrogen Zellumuhle 10 1500Xg 10 4 10 000Xg 10 4 C 0 45um Bio Rad
15. Profinia 5 1 ml O IMAC Bio Rad 1ml GST Bio Rad Profinia 6 SDS PAGE Profinia 10ul 30uD 40ul 95C 5 Bio Rad TGX 4 15 200V 30 ORIOLE Bio Rad 90 7 SDS PAGE 200mA 60 PVDF HA Open Biosystems Invitrogen 3 Western Breeze
16. 3 GCYI KU YPRI Profinia A CCY7 B YPRI M 1A 1B 1C 1D Profinia N N N N N NY NY NY Y JV AM uon 3 PA D ph p Z AN zi 2 P P S NR SR aw KXXX S Gv 3 x NON X X X 44 QS uu X9 XN a ZO GN eV AV AT NE AY c SY aW ue LS S LA IA a QS uM NN S ow TA AS OA a Lu A SOM qd aV xx 43 X TX LEE J 4 CCY7 YPRI A CCY7 B YPRI 100 1D 6 XHis GCY1 YPR1 6 XHis GCYl YPRI Ni 4
17. 236 31 30 Link T Lohaus G Heiser I Mendgen K Hahn M and Voegele RT 2005 Characterization of a novel NADP dependent D arabitol dehydrogenase from the plant pathogen Uromyces fabae Biochem J 389 289 295 Minamihara T and Suzuki M 1997 Properties of sorbitol dehydrogenase from Pseudomonas sp KS E1806 and comparison with other sorbitiol dehydrogenases J Ferment Bioeng 84 254 256 Oliveira HM Segundo MA et al 2006 Kinetic enzymatic determination of glycerol in wine and beer using a sequential injection system with spectrophotometric detection J Agric Food Chem 54 4136 4140 Onishi H 1963 Osmophillic yeast Adv Food Res 12 53 mor VE 39 Y Stein R Gross W and Schnarrenberger C 1997 Characteri zation of a xylitol dehydrogenase and a D arabitol dehydro genase from the thermo and acidophilic red alga Galdieria sulphuraria Planta 202 487 493 Tang XM Kayingo G and Prior BA 2005 Functional analysis of the Zygosaccharomyces rouxii Fpslp homo logue Yeast 22 571 81 van Zyl PJ and Prior BA 1990 Water relations of polyol accumulation by Zygosaccharomyces rouxii in continuous culture Appl Microbiol Biotechnol 33 12 17 van Zyl PJ Kilian SG and Prior PA 1990 The role of an active transport mechanism in glycerol accumulation during osmoregulation by Zygosaccharomyces rouxii Appl Micro biol Biotechnol 34 231 235 Watanabe Y Nagayama K a
18. Watanabe et al 2008 Zygosaccharomyces rouxii Saccharomyces cerevisiae DZ Iwaki et al 2001 Z rouxi van Zyl et 34 E XL WE HA al 1990 Tang et al 2005 de Figueroa and Lucca 2001 Lb BEBE S cerevisiae ORF YEL070w Watanabe et aL 2006 2008
19. EE momo m k Jd r L me mm i i i i 1 100 120 140 120 140 m 1 His Ni Profinia A B CCY7 C YPRI 1IA 1B 1C 1D UV1 280nm Ni 140 160mD UV 2 280nm Profinia IMAC UV280nm 1 1A 1B 1C 1D 1 A 1D 1 B CCY7 GCY1 1D
20. MS Microbiol Lett 252 35 42 Costenoble R Valadi H Gustafsson L Niklasson C and Franzen CJ 2000 Microaerobic glycerol formation in Saccharomyces cerevisiae Yeast 16 1483 1495 Saccharomyces cerevisiae de Figueroa LIC and Lucca ME 2001 Prodction of polyols by omsmotolerant yeasts Food Microbiol Protocols 14 99 324 Dominguez KBH Toth IV et al 2010 Sequential injection kinetic flow assay for monitoring glycerol in a sugar fermentation process by Saccharomyces cerevisiae Appl Biochem Biotechnol 160 1664 1673 Dujon B et al 2004 Genome evolution in yeasts Nature 430 6995 35 44 Gelperin DM White MA et al 2009 Biochemical and genetic analysis of the yeast proteome with a movable ORF collection Genes Developm 19 2816 2826 Hallborn J Walfridsson M Penttila M Keranen S and Hahn Hagerdal B 1995 A Short chain dehydrogenase gene from Pichia stipites having D arabitoldehydrogenase activ ity Yeast 11 839 847 Iwaki T Kurono S Yokose Y Kubota K Tamai Y and Watanabe Y 2001 Cloning of glycerol 3 phosphate dehydrogenase gene ZrGPDI and ZrGPD2 and glycerol dehydrogenase genes ZrGCY and ZrGCY2 from the salt tolerant yeast Zygosaccharomyces rouxii Yeast 18 1791 144 Lin ECC 1961 An inducible D arabitol dehydrogenase from Aerobacter aerogenes J Biol Chem
21. nd Tamai Y 2008 Expession of glycerol 3 phosphate dehydrogenase gene CvGPDI in salt tolerant yeast Candida versatilis in stimulated by high concentrations of NaCl Yeast 25 107 116 Watanabe Y Takechi Y Nagayama K and Tamai Y 2006 Overexpression of Saccharomyces cerevisiae mannitol dehydrogenase gene YELO7Ow in glycerol synthesis deficient S cerevisiae mutant Enzyme Microbial Technol 39 654 659 Watanabe Y Tsuchimoto S and Tamai Y 2004 Heterolo gous expression of Zygosaccharomyces rouxii glycerol 3 phosphate dehydrogenase gene ZrGPDI and glycerol dehydrogenase gene ZrGCYI1 in Saccharomyces cere visiae FEMS Yeast Res 4 505 510 Wong B Murray JS Castellanos M and Croen KD 1993 D arabitol metabolism in Candida albicans Studies of the biosynthetic pathway and the gene that encodes NAD dependent D arabitol dehydrogenase J Bacteriol 175 6314 6320 EAT 2008 Food Function 4 1 6
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