富核糖核酸热带假丝酵母发酵培养基优化
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摘要
以热带假丝酵母(Candida tropicalis)AY91009为试验菌株,以其核糖核酸(RNA)含量为目标,选取糖蜜为碳源,采用响应面法对其发酵培养基进行优化,建立酵母浸粉、NH_4Cl和ZnSO_4·7H_2O的二次回归模型,确定培养基最佳配方为:糖蜜(30%含糖量)140 mL/L、酵母浸粉2.90%、NH_4Cl 1.37%、NaH_2PO_4·2H_2O 0.10%、MgSO_4·7H_2O 0.20%、FeSO_4·7H_2O 0.05%、ZnSO_4·7H_2O 0.10%。在此优化培养基中发酵培养12 h,RNA含量达到11.58%,比优化前提高了35.6%。20 L罐分批发酵试验结果表明,细胞干质量达到32.38 g/L,RNA含量为6.69%,总RNA含量为2.17 g/L,为后续的高密度发酵研究奠定了良好的基础。
Using Candida tropicalis AY91009 as experimental strain, the RNA content as the target, and molasses as the carbon source, the fermentation medium was optimized by response surface methodology, and a quadratic regression model with yeast extract, NH_4Cl and ZnSO_4·7H_2O as independent variables was established. The optimum medium formula was determined as follows: molasses(sugar content 30%) 140 ml/L, yeast extract 2.90%, NH_4Cl 1.37%, NaH_2PO_4·2 H_2O 0.10%, Mg SO_4·7 H_2O 0.20%, FeSO_4·7 H_2O 0.05% and ZnSO_4·7 H_2O 0.10%. The RNA content reached 11.58%under the optimum medium fermented for 12 h, which was 35.6% higher than that before optimization. The results of batch fermentation experiment in 20 L fermenter showed that the dry cell mass reached 32.38 g/L, the RNA content was 6.69%, and the total RNA content was 2.17 g/L, which laid a good foundation for the subsequent high-density fermentation research.
Using Candida tropicalis AY91009 as experimental strain, the RNA content as the target, and molasses as the carbon source, the fermentation medium was optimized by response surface methodology, and a quadratic regression model with yeast extract, NH_4Cl and ZnSO_4·7H_2O as independent variables was established. The optimum medium formula was determined as follows: molasses(sugar content 30%) 140 ml/L, yeast extract 2.90%, NH_4Cl 1.37%, NaH_2PO_4·2 H_2O 0.10%, Mg SO_4·7 H_2O 0.20%, FeSO_4·7 H_2O 0.05% and ZnSO_4·7 H_2O 0.10%. The RNA content reached 11.58%under the optimum medium fermented for 12 h, which was 35.6% higher than that before optimization. The results of batch fermentation experiment in 20 L fermenter showed that the dry cell mass reached 32.38 g/L, the RNA content was 6.69%, and the total RNA content was 2.17 g/L, which laid a good foundation for the subsequent high-density fermentation research.
引文
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[2]GRIMBLE G K.Why are dietary nucleotides essential nutrients[J].Brit JNutr,1996,76(4):475-478.
[3]张志军,温明浩,王克文,等.核苷酸生产技术现状及展望[J].现代化工,2004(11):19-23.
[4]CHOUDHURY D,PAL A K,SAHU N P,et al.Dietary yeast RNA supplementation reduces mortality by Aeromonas hydrophila in rohu(Labeo rohita L.)juveniles[J].Fish Shellfish Immun,2005,19(3):281-291.
[5]朱才庆,徐燚,应汉杰,等.核糖核酸的应用与制备[J].南京化工大学学报(自然科学版),2001(04):97-100.
[6]SINGH R,GOPALAN S,SIBAL A.Immunonutrition[J].Indian J Pediatr,2002,69(5):417-419.
[7]邱蔚然,丁庆豹.核酸类产品的生物合成[M].上海:华东理工大学出版社,2006:56-73.
[8]万文军,罗众球,邱蔚然,等.酵母发酵法制备核糖核酸研究进展[J].生物技术通讯,2008,19(4):638-640.
[9]张子健,江建梅,舒媛,等.高核糖核酸啤酒酵母菌QH633菌体制备工艺优化[J].食品工业科技,2015,36(3):178-181.
[10]SHINJI Y,HISAO K,TAIJU M.Ribonucleic acid-enriched brewer's yeast cells and process for producing the same:US,7135327 B2[P].2006-11-14.
[11]NAGODAWITHANA T.Yeast-derived flavors and flavor enhancers and their probable mode of action[J].Food Technol,1992,46(12):138-144.
[12]YOSHIO N,ISAO B,et al.Process for preparing yeast cells containing an enhanced amount of ribonucleic acid:US 3411989 A[P].1968-11-19.
[13]杨依顺,丁玥,曹静,等.酿酒酵母发酵制备RNA的研究[J].药物生物技术,2013,20(4):310-313.
[14]王陶,王伟,周卫东,等.废糖蜜培养高核酸酵母提取RNA的研究[J].中国食品工业,2010(11):82-84.
[15]张秀凤,辛嘉英,畅乐.红酵母发酵废糖蜜产类胡萝卜素的研究[J].食品工业科技,2013,34(1)184-187.
[16]王媛媛,曹春红,杨旭,等.糖蜜原料培养高核酸酵母NY-1培养条件优化[J].食品与发酵工业,2012,38(1):62-65.
[17]LIU Y P,ZHENG P,SUN Z H,et al.Economical succinic acid production from cane molasses by Actinobacillus succinogenes[J].Bioresource Technol,2008,99(6):1736-1742.
[18]盛建国,李灵敏.K-79高核酵母发酵废糖蜜制备核糖核酸的研究[J].食品工业科技,2007,28(1):140-142.
[19]SOFIA H,JOHAN P,THOMAS A,et al.Growth rate and medium composition strongly affect folate content in Saccharomyces cerevisiae[J].Int J Food Microbiol,2008,123(1-2):93-100.
[20]俞灿,郑国斌,姚娟,等.富含核糖核酸酿酒酵母的选育及其高密度发酵工艺[J].中国酿造,2016,35(12):66-71.
[21]MURALIDHAR R V,CHIRUMAMILA R R,MARCHANT R,et al.Aresponse surface approach for the comparison of lipase production by Canida cylindracea using two different carbon sources[J].Biochem Eng J,2001,9(1):17-23.
[22]YE D,XU Z N,CEN P L.Medium optimization for enhanced production of cytosine-substituted mildiomycin analogue(MIL-C)by Streptoverticillium rimofaciens ZJU 5119[J].J Zhejiang Univer,2008,9(1):77.
[23]陈文明,郑国斌,姚娟,等.YE专用高蛋白高RNA酵母菌株的筛选及鉴定[J].食品科技,2015,40(4):2-6.