酪丁酸梭菌RL1产丁酸发酵条件优化研究
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摘要
以从浓香型白酒窖泥中分离得到的酪丁酸梭菌RL1(Clostridium tyrobutyricum RL1)为研究对象,采用单因素试验结合正交试验优化菌株产丁酸的发酵条件.结果表明,酪丁酸梭菌RL1的最优产丁酸发酵条件为:发酵培养基初始pH=6. 8,装液量100%,培养温度34℃,接种量3%(体积分数),还原剂为硫代乙醇酸钠且其添加量为0. 5 g/L.在该发酵条件下,酪丁酸梭菌RL1的丁酸产量可达10. 66 g/L,较优化前提高了41. 76%.
The fermentation conditions of Clostridium tyrobutyricum RL1 isolated from Luzhou-flavor liquor pit mud for the production of butyric acid was optimized using one-factor experiment combined with orthogonal experiment. The results showed that the optimal yield of butyric acid termentation of Clostridium butyrium RL1 was the initial p H value 6. 8,the volume of culture media 100%,the culture temperature 34 ℃,the inoculation amount 3%( volume fraction) and the dosage of the deoxidizer( sodium thioglycolate) 0. 5 g/L. Under the optimal fermentation conditions,the production of butyric acid by Clostridion tyrobutyricum RL1 reached10. 66 g/L,which was 41. 76% higher than that before optimization.
The fermentation conditions of Clostridium tyrobutyricum RL1 isolated from Luzhou-flavor liquor pit mud for the production of butyric acid was optimized using one-factor experiment combined with orthogonal experiment. The results showed that the optimal yield of butyric acid termentation of Clostridium butyrium RL1 was the initial p H value 6. 8,the volume of culture media 100%,the culture temperature 34 ℃,the inoculation amount 3%( volume fraction) and the dosage of the deoxidizer( sodium thioglycolate) 0. 5 g/L. Under the optimal fermentation conditions,the production of butyric acid by Clostridion tyrobutyricum RL1 reached10. 66 g/L,which was 41. 76% higher than that before optimization.
引文
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[2] WEIMER P J,STEVENSON,D M. Isolation characterization and quantification of Clostridium kluyveri from the bovine rumen[J]. Appl Microbiol Biotechnol,2012,94(2):461.
[3] WANG J,LIN M,XU M,YANG S T. Anaerobic fermentation for production of carboxylic acids as bulk chemicals from renewable biomass[J].Adv Biochem Eng Biotechnol,2016,156:323.
[4] LUO H,GE L,ZHANG J,et al. Enhancing butanol production under the stress environments of co-culturing Clostridium acetobutylicum/Saccharomyces cerevisiae integrated with exogenous butyrate addition[J]. Plos One,2015,10(10):e0141160.
[5] LUO H,ZENG Q,HAN S,et al. High-efficient n-butanol production by co-culturing Clostridium acetobutylicum and Saccharomyces cerevisiae integrated with butyrate fermentative supernatant addition[J]. World J Microbiol Biotechnol,2017,33(4):76.
[6] RICHTER H,QURESSHI N,HEGER S,et al.Prolonged conversion of n-butyrate to n-butanol with Clostridium saccharoperbutylacetonicum in a two-stage continuous culture with in-situ product removal[J]. Biotechnology&Bioengineering,2012,109(4):913.
[7] TASHIRO Y,TAKEDA K,KOBAYASHI G,et al. High butanol production by Clostridium saccharoperbutylacetonicum N1-4 in fed-batch culture with p H-stat continuous butyric acid and glucose feeding method[J]. Journal of Bioscience&Bioengineering,2004,98(4):263.
[8]周丽春.米糠纤维床反应器固定化发酵产丁酸的研究[D].杭州:浙江大学,2016.
[9] MAITI S,BRAR S K,VERMA M,et al. Chapter7—butyric acid:A platform chemical for biofuel and high-value biochemicals[M]∥Brar S K,Sarma S J,Pakshirajan K. Platform Chemical Biorefinery. Amsterdam:Elsevier Inc,2016:119.
[10] YANG X,TANG S,LU T,et al. Chem Inform abstract:sulfonic acid resin—Catalyzed oxidation of aldehydes to carboxylic acids by hydrogen peroxide[J]. Journal of Energy Chemistry,2013,44(4):659.
[11] IVAN B,PETER W. Microbial production of short chain fatty acids from lignocellulosic biomass:Current processes and market[J].Biomed Research International,2016(1):8469357.
[12] REPHAELI A,ZHUK R,NUDELMAN A. Prodrugs of butyric acid from bench to bedside:Synthetic design mechanisms of action and clinical applications[J]. Drug Development Research,2010,50(3/4):379.
[13] ZIGOVA J,STURAIK E. Advances in biotechnological production of butyric acid[J]. Journal of Industrial Microbiology&Biotechnology,2000,24(3):153.
[14]邓名荣,郭俊,朱红惠.微生物催化生产丁酸研究进展[J].中国生物工程杂志,2009,29(3):117.
[15]江凌.纤维床固定化酪丁酸梭菌发酵廉价生物质生产丁酸的研究[D].广州:华南理工大学,2010.
[16]何培新,李聪聪,胡晓龙,等.基于HS-SPMEGC-MS的浓香型白酒窖泥中可培养Clostridium spp.挥发性代谢物成分分析[J].轻工学报,2017,32(6):1.
[17] HU X L,DU H,XU Y. Identification and quantification of the caproic acid-producing bacterium Clostridium kluyveri in the fermentation of pit mud used for Chinese strong-aroma type liquor production[J]. International Journal of Food Microbiology,2015,214:116.
[18]于平,陈凯飞,朱祺,等.重组大肠杆菌生物合成γ-氨基丁酸的发酵条件优化[J].中国食品学报,2018(6):112.
[19]陈兴杰,吴攀攀,徐敏瑞,等.窖泥己酸菌和丁酸菌联合接种发酵液有机酸检测与分析[J].酿酒,2017,44(3):60.
[20]项宜娟.纤维床固定化酪丁酸梭菌产丁酸的研究[D].杭州:浙江理工大学,2010.
[21]高振,江凌,朱丽英,等.纤维床固定化酪丁酸梭菌发酵大薯渣水解液生产丁酸的研究[J].安徽农业科学,2012,40(33):16302.