牛肝菌胞外多糖发酵培养基的优化

在Plackett Burman设计实验结果基础上 ,采用响应曲面法对影响牛肝菌 (Boletussp .)ACCC 5 0 3 2 8发酵胞外多糖的培养基 5个关键组成成分酵母膏 (X1)、麦芽糖 (X2 )、(NH4 ) 2 SO4(X3)、FeSO4 (X4 )和CuSO4 ·5H2 O(X5)的最佳水平范围进行了研究和探讨。通过对二次多项回归方程求解得知 ,在上述自变量取值分别为 :酵母膏 1 4 .2 g/L ,麦芽糖 2 2 .2g/L ,(NH4 ) 2 SO4 2 .7g/L ,FeSO4 66.9mg/L ,CuSO4 1 0 1 .5 9μg/L时 ,胞外多糖最大预测值为 75 2 .0 79μg/mL(发酵醪 ) ,此预测可信度不仅被统计分析所验证 ,也被实践所证实     

牛肝菌摇瓶发酵条件优化研究

在Plackett Burman实验结果基础上 ,采用响应曲面法 (responsesurfacemethodology ,RSM )对影响牛肝菌 (Boletussp )ACCC 5 0 3 2 8发酵产糖与生长的关键工艺参数 ,发酵温度、发酵时间和装液量的最佳水平范围进行了研究和探讨。通过对胞外多糖曲面方程以及菌丝干重二次多项回归方程求解得知 ,在自变量发酵温度 ,发酵时间和装液量分别为 2 4 2 3℃、1 0 7d和 5 0 0mL时 ,牛肝菌ACCC 5 0 3 2 8菌株发酵胞外多糖的最大预测值为 1 0 5 3 99μg/mL发酵醪 ;在 2 6 77℃、1 0 9d和 64 90mL时 ,该菌干菌丝浓度可达 4 2 2mg/mL发酵醪。欲同时获得最大量的胞外多糖和菌丝浓度 ,则发酵温度 ,发酵时间和装液量须为 2 6 2 3℃、1 0 5d和 5 8 70mL ,在此条件下 ,每毫升发酵醪中的胞外多糖含量和菌丝浓度分别为 992 2 6μg与 4 1 6mg。上述预测值不仅被统计学方法所验证 ,而且也被后续实验所证实     

酵母发酵法分离纯化低聚异麦芽糖的研究

酿酒酵母As2.109经驯化后,可用于发酵法分离纯化低聚异麦芽糖。优化后酵母发酵的最佳条件为:pH4.0,温度30～32℃,低聚异麦芽糖浓度250g/L,酵母膏浓度0.6g/L,CO(NH2)20.6g/L,Fe2(SO4)30.08g/L,MgSO41.2g/L。在此条件下,低聚异麦芽糖经40h发酵后,纯度由54.84%提高到90.83%,而其中的葡萄糖被完全除去。     

响应面法优化红曲米中凝乳酶高产菌株的发酵条件

对从红曲米中分离得到的产凝乳酶能力强的菌株M5传代菌株的液态发酵培养基及产酶条件进行优化。首先进行单因素实验得到适宜的氮源为(NH4)2SO4、酪蛋白,无机盐为FeSO4、KH2PO4,适宜的接种量为1%,培养温度为30℃,培养时间为5d,发酵培养基初始pH为6.0。在此基础上通过Plackett-Burman实验筛选出对酶活影响显著的三个因素:(NH4)2SO4含量、培养时间和培养温度。再用Box-Behnken响应曲面实验对三个显著因素进行优化。结果表明,产酶的最佳培养基组分:(NH4)2SO40.53%、FeSO40.05‰、KH2PO40.05%、干酪素0.5%、葡萄糖2%、接种量1%。最佳发酵条件为:培养温度31.3℃、摇床转速为180r/min、培养时间113h、pH6.0。基于响应曲面优化的产凝乳酶培养基组成与发酵条件效果显著,供试菌株M5传代菌株所产凝乳酶活性由45.34SU/mL提高到190.68SU/mL。     

酵母发酵法分离纯化低聚异麦芽糖的研究

酿酒酵母As2109经驯化后,可用于发酵法分离纯化低聚异麦芽糖。优化后酵母发酵的最佳条件为：pH4．0,温度30－32℃,低聚异麦芽糖浓度250g/L,酵母膏浓度．6g/L,CO(NH2)20.6G/l,Fe(SO4)30.08g/L,MgSO41.2g/L。在此条件下,低聚异麦芽糖经38h发酵后,低聚异麦芽糖纯度由53．8％提高到90．3％,而葡萄糖被完全除去。     

水溶性红曲色素液态发酵培养基成分优化

对ACCC 30352(GIM 3.439)紫红曲霉液态发酵培养基成分进行筛选,发现葡萄糖、蔗糖、麦芽糖、可溶性淀粉、有机氮源、Mg2+、Zn2+、Mn2+、Fe2+、甘氨酸有利于色素合成;最适碳源含量为5%;最适碳氮比(质量比)为1:1。在此基础上,通过正交试验对碳源、氮源和碳氮比(质量比)作进一步分析,确定其液态发酵代谢水溶性色素的最佳培养基成分为:蔗糖5%、酵母膏1.07%、Na NO3 0.1%、甘氨酸0.5%、Mg SO4 0.05%、Zn SO4 0.001%、Mn SO4 0.003%、Fe SO40.001%。     

均匀设计法优化柞蚕生产用Avermectins的发酵培养基

本实验以Streptomyces avermitilis为实验菌株,通过均匀设计方法优化了柞蚕生产用avermectins发酵培养基.结果表明:以玉米淀粉9%、黄豆饼粉0.5%、花生饼粉0.7%、酵母粉0.8%、酵母膏0.1%、玉米浆0.3%、CoCl2*6H2O 0.003%、(NH4)2SO4 0.02%的培养基配方可使avermectins发酵产量最高.     

培养基中各营养组分对酿酒酵母发酵的影响

酿酒酵母常被用于乙醇发酵,作为能量和营养的来源,培养基的组分对于酿酒酵母的生长以及耐酒精能力有很大的影响,使用Plackett-Burman设计方法,通过对酿酒酵母发酵培养基中的各个组分进行分析研究,发现各个组分的影响力大小为:酵母膏>蛋白胨>MgSO4>CaCl2>(NH4)2SO4>KH2PO4。通过单独对酵母膏和蛋白胨在酵母细胞生长及耐酒精性能方面的研究,发现提高其含量,可以在发酵的各个阶段提高酵母细胞的细胞密度、乙醇耐受性及发酵能力。     

浓香型白酒生产中耐酒精酵母菌的选育研究

以宋河酒厂窖泥、酒糟、酒厂废水为样品,从中分离出100株菌株,经初筛、复筛和酒精发酵试验,最终获得1株耐酒精度为18%vol酵母菌,编号为A16。通过单因素实验确定最佳发酵条件为:发酵温度30℃、p H值4.5、碳源为葡萄糖、氮源为复合氮源(酵母膏和蛋白胨的比例为2∶1)、无机盐为Mg SO4·7H2O、接种量为10%、转速100 r/min、装液量为150 m L/250 m L;通过正交试验确定培养基最佳配比为:葡萄糖25%,酵母膏2%,蛋白胨1%,Mg SO4·7H2O 1%,(NH4)2SO41%,KH2PO41%,菌株A16的酒精得率70%。     

基于胞外多糖和菌丝生物量的香菇发酵培养基优化

为研究液体发酵培养基对香菇胞外多糖和菌丝生物量的影响,以秦巴山区香菇808菌株为试材,采用Plackett-Burman设计实验、最陡爬坡实验和响应曲面法对其液体发酵培养基的碳源、氮源和其它营养物质进行优化。结果表明,香菇胞外多糖发酵培养基的最佳组合是(g/500 m L):蔗糖7.18,玉米粉15.00,麦麸14.05,酵母膏0.35,KH2PO40.50,Mg SO4·7H2O 0.50,p H自然,胞外多糖实测值为0.967 g/500 m L;香菇菌丝生物量发酵培养基的最佳组合是(g/500 m L):蔗糖7.18,玉米粉15.00,麦麸14.05,酵母膏0.35,KH2PO40.75,Mg SO4·7H2O 0.50,p H自然,菌丝生物量实测值为28.146 g/500 m L。优化后的香菇胞外多糖产量和菌丝生物量较优化前分别提高12.44%和11.00%。此研究结果可为香菇液体发酵的中试生产提供理论依据。     

呕吐毒素脱毒菌Devosia sp.发酵培养基优化

德沃斯氏菌(Devosia sp.)是一种可以高效脱除谷物及饲料中呕吐毒素的新型微生物菌株,其转化产物是目前研究较为安全的呕吐毒素转化产物。本研究以碳、氮源单因素实验为基础,选用8个因素进行部分因子试验(FFD),筛选出酵母浸粉、酵母膏和蔗糖3个最显著影响因素;对3个最显著影响因素进行最陡爬坡实验获得其最优点区域;再通过中心组合实验设计(CCD)进行响应面分析得到了Devosia sp.的最适发酵培养基组成为:酵母浸粉10.00 g/L,酵母膏6.49 g/L,蔗糖8.40 g/L,硫酸铵2 g/L,磷酸二氢钾2 g/L,硫酸镁0.7 g/L,氯化钙0.02 g/L和硫酸亚铁0.02 g/L。使用该培养基进行发酵验证试验,培养40 h的OD₆₀₀达到21.66,与预测值(21.62)基本相符。综上,该回归模型对Devosia sp.发酵培养基的优化是可行的,且优化后较原始培养基生物量(OD₆₀₀)提高了89.1%,降低了生产成本,为呕吐毒素脱毒菌株的工业生产奠定了基础。     

酱油曲霉发酵芝麻饼粕的条件优化

采用响应面法对酱油曲霉固体发酵芝麻饼粕提高抗氧化能力的条件进行优化。通过Plackett-Burman设计法,评价麦芽糖、葡萄糖、蔗糖、果糖、麦麸、玉米浆、酵母膏和硫酸铵8个因素对清除DPPH自由基能力的影响。筛选出麦麸、葡萄糖和硫酸铵为影响芝麻饼粕发酵提取物抗氧化活性的3个显著因素,然后采用中心组合试验设计和响应面分析法,对3个关键因素进行优化。优化的发酵条件为:葡萄糖、硫酸铵和麦麸添加量分别为1.87%、1.54%和2.55%。在该实验条件下,DPPH自由基清除活性为80.26%。     

氮源对L-色氨酸发酵的影响

以L-色氨酸生产菌E.coli TRTH为供试菌株,研究了氮源对L-色氨酸发酵的影响。利用30 L发酵罐进行分批补料发酵试验,确定了最佳有机氮源和无机氮源分别为酵母粉和硫酸铵,进一步确定酵母粉和硫酸铵的最佳用量为1 g/L和10 g/L,最后采用NaOH和氨水混合补料控制发酵液中NH4+浓度在120 mmol/L以下,发酵38 h,菌体生物量和L-色氨酸产量分别达到53.42 g/L和32.6 g/L,实现了大肠杆菌的高密度培养。     

Structural Complexity of the Nitrogen Source and Influence on Yeast Growth and Fermentation

The structural complexity of the nitrogen source strongly affects both biomass and ethanol production by industrial strains of Saccharomyces cerevisiae, during fermentation in media containing glucose or maltose, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low glucose and maltose concentrations independent of nitrogen supplementation. At high sugar concentrations diauxie was not easily observed, and growth and ethanol production depended on the nature of the nitrogen source. This was different for baking and brewing ale and lager yeast strains. Sugar concentration had a strong effect on the shift from oxido‐fermentative to oxidative metabolism. At low sugar concentrations, biomass production was similar under both peptone and casamino acid supplementation. Under casamino acid supplementation, the time for metabolic shift increased with the glucose concentration, together with a decrease in the biomass production. This drastic effect on glucose fermentation resulted in the extinction of the second growth phase, probably due to the loss of cell viability. Ammonium salts always induced poor yeast performance. In general, supplementation with a nitrogen source in the peptide form (peptone) was more positive for yeast metabolism, inducing higher biomass and ethanol production, and preserving yeast viability, in both glucose and maltose media, for baking and brewing ale and lager yeast strains. Determination of amino acid utilization showed that most free and peptide amino acids present, in peptone and casamino acids, were utilized by the yeast, suggesting that the results described in this work were not due to a nutritional status induced by nitrogen limitation.     

皮状丝孢酵母发酵菊粉产油脂培养基的优化

采用Plaekett-Burman设计和响应面分析法,对皮状丝孢酵母菌发酵菊粉产油脂的培养基进行优化.首先用Plaekett-Burman设计对发酵培养基组分菊粉、酵母膏、硝酸钾、硫酸镁、硫酸铁、磷酸二氢钾、氯化钙、硫酸锌等进行评价并筛选出了有显著效应的酵母膏、硫酸锌、氯化钙,然后用最陡爬坡路径逼近最大响应值区域,最后再利用Box-Behnken试验设计及响应面分析法进行回归分析.结果表明酵母膏、硫酸锌、氯化钙于油脂含量存在显著的相关性.通过求解回归方程得到优化发酵培养基组分:酵母膏6.68g/L、硫酸锌0.457g/L、氯化钙0.342g/L.对优化后的培养基进行发酵培养,其生物量和油脂含量分别达到了13.56g/L、5.281g/L,油脂含量比优化之前增加了11.2％.     

Altered Patterns of Maltose and Glucose Fermentation by Brewing and Wine Yeasts Influenced by the Complexity of Nitrogen Source

Maltose and glucose fermentations by industrial brewing and wine yeasts strains were strongly affected by the structural complexity of the nitrogen source. In this study, four Saccharomyces cerevisiae strains, two brewing and two wine yeasts, were grown in a medium containing maltose or glucose supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low sugar concentration for brewing and wine strains, independent of nitrogen supplementation, and the type of sugar. At high sugar concentrations altered patterns of sugar fermentation were observed, and biomass accumulation and ethanol production depended on the nature of the nitrogen source and were different for brewing and wine strains. In maltose, high biomass production was observed under peptone and casamino acids for the brewing and wine strains, however efficient maltose utilization and high ethanol production was only observed in the presence of casamino acids for one brewing and one wine strain studied. Conversely, peptone and casamino acids induced higher biomass and ethanol production for the two other brewing and wine strains studied. With glucose, in general, peptone induced higher fermentation performance for all strains, and one brewing and wine strain produced the same amount of ethanol with peptone and casamino acids supplementation. Ammonium salts always induced poor yeast performance. The results described in this paper suggest that the complex nitrogen composition of the cultivation medium may create conditions resembling those responsible for inducing sluggish/stuck fermentation, and indicate that the kind and concentration of sugar, the complexity of nitrogen source and the yeast genetic background influence optimal industrial yeast fermentation performance.     

Effect of Sugar Catabolite Repression in Correlation with the Structural Complexity of the Nitrogen Source on Yeast Growth and Fermentation

Biomass and ethanol production by industrial Saccharomyces cerevisiae strains were strongly affected by the structural complexity of the nitrogen source during fermentation in media containing galactose, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low galactose concentrations independent of nitrogen supplementation. At high sugar concentrations altered patterns of galactose utilisation were observed. Biomass accumulation and ethanol production depended on the nature of the nitrogen source and were different for baking and brewing ale and lager strains. Baking yeast showed improved galactose fermentation performance in the medium supplemented with casamino acids. High biomass production was observed with peptone and casamino acids for the ale brewing strain, however high ethanol production was observed only in the presence of casamino acids. Conversely, peptone was the nitrogen supplement that induced higher biomass and ethanol production for the lager brewing strain. Ammonium salts always induced poor yeast performance. The results with galactose differed from those obtained with glucose and maltose which indicated that supplementation with a nitrogen source in the peptide form (peptone) was more positive for yeast metabolism, suggesting that sugar catabolite repression has a central role in yeast performance in a medium containing nitrogen sources with differing levels of structural complexity.     

响应面法优化枯草芽孢杆菌3-羟基丁酮发酵培养基

采用响应面法对枯草芽孢杆菌TH-55 3-羟基丁酮发酵培养基进行了优化,确定了葡萄糖、酵母浸膏和玉米浆是影响菌株3-羟基丁酮发酵产率的主要因素.优化获得的发酵培养基组成:葡萄糖102g/L,酵母浸提物6.8g/L,玉米浆26.5g/L,硫酸铵5g/L,硫酸锰0.05g/L,硫酸镁0.6g/L,磷酸二氢钾0.5g/L.在此条件下,菌株摇瓶发酵3-羟基丁酮平均产率达到46.25g/L,较优化前的菌株产率35.21g/L相比提高了31％.10L发酵罐发酵试验,发酵周期72h,3-羟基丁酮发酵产率达到47.85g/L.