球头三型孢菌产赤藓糖醇的两阶段pH值调控
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摘要
为了进一步提高球头三型孢菌发酵生产赤藓糖醇的产量,研究了不同pH值对菌体生长、赤藓糖还原酶(ER)酶活和赤藓糖醇合成的影响。结果表明,单一pH值控制不能使菌体生物量和赤藓糖醇产量同时达到最优值。通过分析不同pH值下发酵过程曲线以及ER酶活变化,提出了两阶段pH值调控策略,即在发酵前期(0～36 h)控制pH=5.0,之后控制pH=6.0。通过采用这一优化策略,发酵过程中的最大ER酶活为0.55 U·mg~(-1) protein,比单一控制pH值策略的最大ER酶活(pH=6.0,0.38 U·mg~(-1) protein)提高了44.74%。发酵120 h后,赤藓糖醇的产量达到56.78 g·L~(-1),比单一控制pH值优化的最大产量(pH=6.0,46.75 g·L~(-1))提高了21.45%。
In order to further improve the yield of erythritol produced by Trichosporonoides oedocephalis,we studied the effect of different pH values on cell growth,erythrose reductase(ER) activity and erythritol biosynthesis. The results show that mono-control of p H cannot satisfactorily offer higher biomass and erythritol produc-tion simultaneously. Based on time courses of cell growth,activity of ER and erythritol formation,a two-stage pH control strategy was therefore developed,in which pH was controlled at 5.0 for the first 36 h and then 6.0 for the remaining time. Under the optimized strategy,the maximal ER activity in the fermentation process was0.55 U·mg~(-1) protein,which was 44.74% higher than the maximum of mono-control of pH(pH=6.0,0.38 U·mg~(-1) protein). After 120 h of fermentation,the yield of erythritol reached 56.78 g·L~(-1),which was 21.45% higher than that of the maximum of mono-control of pH(pH=6.0,46.75 g·L~(-1)).
In order to further improve the yield of erythritol produced by Trichosporonoides oedocephalis,we studied the effect of different pH values on cell growth,erythrose reductase(ER) activity and erythritol biosynthesis. The results show that mono-control of p H cannot satisfactorily offer higher biomass and erythritol produc-tion simultaneously. Based on time courses of cell growth,activity of ER and erythritol formation,a two-stage pH control strategy was therefore developed,in which pH was controlled at 5.0 for the first 36 h and then 6.0 for the remaining time. Under the optimized strategy,the maximal ER activity in the fermentation process was0.55 U·mg~(-1) protein,which was 44.74% higher than the maximum of mono-control of pH(pH=6.0,0.38 U·mg~(-1) protein). After 120 h of fermentation,the yield of erythritol reached 56.78 g·L~(-1),which was 21.45% higher than that of the maximum of mono-control of pH(pH=6.0,46.75 g·L~(-1)).
引文
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[15]KOBAYASHI Y,YOSHIDA J,IWATA H,et al.Gene expression and function involved in polyol biosynthesis of Trichosporonoides megachiliensis under hyper-osmotic stress[J].Journal of Bioscience and Bioengineering,2012,115(6):645-650.
[16]JOVANOVIC'B,MACH R L,MACH-AIGNER A R.Characterization of erythrose reductases from filamentous fungi[J].AMB Express,2013,3(1):1-11.
[2]JOVANOVIC'B,MACH R L,MACH-AIGNER A R.Erythritol production on wheat straw using Trichoderma reesei[J].AMB Express,2014,4(1):34-45.
[3]LIVESEY G.Health potential of polyols as sugar replacers,with emphasis on low glycaemic properties[J].Nutrition Research Reviews,2003,16(2):163-191.
[4]HASHINO E,KUBONIWA M,ALGHAMDI S A,et al.Erythritol alters microstructure and metabolomic profiles of biofilm composed of Streptococcus gordonii and Porphyromonas gingivalis[J].Molecular Oral Microbiology,2013,28(6):435-451.
[5]OKU T,NAKAMURA S.Threshold for transitory diarrhea induced by ingestion of xylitol and lactitol in young male and female adults[J].Journal of Nutritional Science and Vitaminology,2007,53(1):13-20.
[6]郑建仙.功能性糖醇[M].北京:化学工业出版社,2005:28-68.
[7]CHIDA S,OCHIAI T.Mutant strain of Trichosporonoides oedocephalis:US5916797[P].1999-06-29.
[8]HARTKE A,BOUCH魪S,GIARD J C,et al.The lactic acid stress response of Lactococcus lactis subsp Lactis[J].Current Microbiology,1996,33(3):194-199.
[9]谢涛,方慧英,诸葛斌,等.产甘油假丝酵母稳定期酸胁迫耐受性研究[J].食品与发酵工业,2009,35(6):28-31.
[10]YUE M,CAO H L,ZHANG J P,et al.Improvement of mannitol production by Lactobacillus brevis mutant 3-A5 based on dual-stage pH control and fed-batch fermentations[J].World Journal of Microbiology and Biotechnology,2013,29(10):1923-1930.
[11]ZHANG X,BAO T,RAO Z M,et al.Two-stage pH control strategy based on the pH preference of acetoin reductase regulates acetoin and 2,3-butanediol distribution in Bacillus subtilis[J].PLoS One,2014,9(3):e91187.
[12]SI Z J,MACHAKU D,WEI P L,et al.Enhanced fed-batch production of pyrroloquinoline quinine in Methylobacillus sp CCTCC M2016079 with a two-stage pH control strategy[J].Applied Microbiology and Biotechnology,2017,101(12):4915-4922.
[13]OOKURA T,AZUMA K,ISSHIKI K,et al.Primary structure analysis and functional expression of erythrose reductases from erythritol-producing fungi(Trichosporonoides megachiliensis SNG-42)[J].Bioscience,Biotechnology,and Biochemistry,2005,69(5):944-951.
[14]PARK E H,LEE H Y,RYU Y W,et al.Role of osmotic and salt stress in the expression of erythrose reductase in Candida magnoliae[J].Journal of Microbiology and Biotechnology,2011,21(10):1064-1068.
[15]KOBAYASHI Y,YOSHIDA J,IWATA H,et al.Gene expression and function involved in polyol biosynthesis of Trichosporonoides megachiliensis under hyper-osmotic stress[J].Journal of Bioscience and Bioengineering,2012,115(6):645-650.
[16]JOVANOVIC'B,MACH R L,MACH-AIGNER A R.Characterization of erythrose reductases from filamentous fungi[J].AMB Express,2013,3(1):1-11.