Screening of Respiration-deficient Saccharomyces cerevisiae Strains with Sugar-and Thermo-tolerances

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1 3 3 Vol3 No3 3 9 Life Science Research June 9 *,,, 535 :, 3, 5- (TTC), 3,, T,, ; (5~5 μ 375~5 μ),, 5% (W/V), 55,, 55% 9% : ; ; ; ; :Q39 + :A :7-77(9) Screening of Respiration-deficient Saccharomyces cerevisiae Strains with Sugar-and Thermo-tolerances LAN Zhao TANG Mei-qiong WEI Ke MA Guang-ting * Intitute of Applied Microbiology of Agriculture Guangxi University Nanning 535 Guangxi China Abstract Thirteen respiration deficiency mutants were separated by using UV and TTC identification medium Saccharomyces cerevisiae strains with sugar-and thermo-tolerances serial number T was selected further The surface of this strain is moist milky small and round the cell is oval budding breeding The ratio of alcohol production and utilization reached to 55% and 9% under the condition of 55 5% sugar Key words Saccharomyces cerevisiae respiration deficiency sugar-resisting thermotolerance screening Life Science Research ~3 [,5] TTC 3 5- [,] TTC [H] TF [3] [H] 3 5- TTC [,7] TF : 9-- : 9--9 : 339 DD5 : 9- * 9- Tel mgt7@sinacom

9 TTC NaCl % W / V 5 5 3 5 55 5 CO [7] % = [ V / V% ml g / ml / CM TTC g ] % TTC [] % = [ C H O H O C H O H O g NH SO g 5 g KH PO 5 g MgSO 7H O 5 g CaCl g NaCl g 5 g ml 5 min 3 g 7 cfuml - 5 W 537 nm

2 9 TTC NaCl % W / V CO [7] % = [ V / V% ml g / ml / CM TTC g ] % TTC [] % = [ C H O H O C H O H O g NH SO g 5 g KH PO 5 g MgSO 7H O 5 g CaCl g NaCl g 5 g ml 5 min 3 g 7 cfuml - 5 W 537 nm cm s CM TTC 3 TTC 3 h TTC Fig The picture of ultraviolet lights mutation TTC selection 3~ h CM TTC % V / V 5 g / L d CO 5 CO W cm 537 nm cm s % 5% 999% s 3% W / V 35 5 CO g / L 7 h T T9 T kg % W / V 5 CO 9 g 3 g 3 g % / % ] % TTC TTC 3 5-

3 3 : TTC Table The results of selection respiration deficiency Saccharomyces cerevisiae by ultraviolet lights mutation and TTC medium No Light upon time Beginning density Streak plate volume Streak plate number Survive germ number Germ number of site in TTC medium Germ number that become white Cause death rate Respiration deficiency selection rate /s / 7 cfuml - /ml /number /number /number /number / % / % generous generous Table Fermentation test of respiration deficiency Saccharomyces cerevisiae Strain T T T3 T T5 T T7 T T9 T T T T3 Carbon dioxide yield / g % 5% 5 3% 3 T T9 L 5% 35 5 % 5% T 5% 3% T T 35 Strain T T9 T Table 3 3 Preliminary screening alcohol fermentation rate under megatemperature and high glucose condition Fermentation temperature / 35 5 Contain amount of glucose W/V / % T 5~5 35~ T 3 5 NaCl 3 5~5 μ 5% 375~5 μ cfuml - kg T 93 g 5~5 T T 7 T 5%~5% 5%

9 T 3 T Fig The colonial morphologic characteristics of T ( 5) Fig3 Individual under microscope of T (microscope photo 5) 5 5 35 3 5 5 5 5 5 3 35 Contain amount of glucose W/V / % 7

fermentation fermentation 35 3 5 5 5 5 5 Contain amount of sodium chloride W/V / % 7 cfuml - 5 35 3 5 5 5 5 5 55 5 Origination 7 cfuml - NaCl T 7 T Fig Different sodium chloride

4 9 T 3 T Fig The colonial morphologic characteristics of T ( 5) Fig3 Individual under microscope of T (microscope photo 5) Contain amount of glucose W/V / % 7 cfuml Fermentation temperature / 7 cfuml - T 5 T Fig Different glucose concentration effect of alcohol Fig5 Different temperature effect of alcohol fermentation fermentation Contain amount of sodium chloride W/V / % 7 cfuml Origination 7 cfuml - NaCl T 7 T Fig Different sodium chloride concentration effect of Fig7 Different concentration effect of alcohol alcohol fermentation fermentation T ~7 h CO T 5% 7 g 3 g % g 73 g 3 7 cfuml - 7~ ~ h 9 h CO 3 7 cfuml cfuml - CO

5 3 : 3 9 h 5 g 5% % 9% 55% 55 9 h kg Carbon dioxide yield / one kg glucose Glucose residue / one kg glucose Fermentation time / h 7 cfuml - T Fig Alcohol fermentation curve of glucose of T Saccharomyces cerevisiae A: Carbon dioxide yield(g); B: Glucose residue(g); C: (g); D: ( 7 cfuml - ); E: 3 [J] LIU Bo XIA Shu-you ZHANG Ying Summary of recent research on the technics of bacteria fermentation to produce alcohol[j] Sichuan Food and Ferme- tation -5 mutated by ultraviolet lights[j] Guangxi Journal of Light In- dustry 5 7- TTC 3 3 Wei-feng GAO Dong tion fermentation of alcohol[j] Liquor-making Science & Te- T 5% [] 7 h 9% Liquor-making Science & Technolosy [5] Xin MAO Zhong-gui ~55 chnolosy [] [J] Industrial Water Treatment 9-3 (References): [] [] [J] FENG You-jun HUANG Yan ZHANG Hui-ming et al Screening of respiration deficiency mutants of yeasts Saccharomyces cerevisiae [3] A B C D E [J] JIN Jian-lin LIU Comparing characteristics of ethanol production by respiratory deficient mutants and wild strain in sacchromyces cerevisiae[j] Microbiology [J] WU Shuai XIAO Dong-guang YUAN Tong-lei et al Selection of Saccharomyces cerevisiae with good osmo- thermo-tolerance and high ethanol-production capacity[j] [J] L 譈 Research progress of high concentra- TTC- [J] HU Zi-bin Normal temperature extraction for the determination of TTC-dehydrogenase activity and its application [7] TTC [J] WANG Mei ZHANG Peng-pai SHUAI Gui-lan et al The application of TTC in yellow wine Saccharomyces cerevisiae selection[j] Liquor Making 5 -

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