不同培养方式对红发夫酵母产虾青素的影响

通过间歇流加培养、指数流加培养、混合碳源-番茄汁流加培养和恒速流加培养等4种培养方式的实验,说明进行流加培养有利于红发夫酵母细胞生长及虾青素的合成。与分批培养相比,所有流加模式下生物量及虾青素产量都有不同程度地提高。其中流加混合碳源-番茄汁获得了最好的实验结果,最终的生物量、虾青素含量和产量分别达到33.71g/L、1106μg/g和36.26mg/L。     

Xanthophyllomyces dendrorhous高细胞密度培养及其虾青素合成——响应pH-stat的底物流加方式的研究

主要研究了采用响应 p H- stat的流加培养模式 ,进行 Xanthophyllomyces dendrorhous高细胞密度培养及虾青素合成。在该补料间歇培养模式中 ,发酵过程 p H的恒定控制采用2 .78mol/ L葡萄糖及 3mol/ L氨水来控制 ,同时起到流加葡萄糖的作用。试验结果表明 ,虾青素积累量于 78h可达 4.0 5mg/ L,菌体干重在第 84h获最大值 1 4 .62 g/ L。与不流加葡萄糖、氨水的间歇培养相比 ,虾青素含量和菌体干重分别增加 2 .89、5.2 2倍。     

优化碳源提升法夫酵母虾青素产量和占类胡萝卜素比例

为进一步提升法夫酵母虾青素产量和虾青素占类胡萝卜素比例,考察了大宗碳源及其组合对法夫酵母细胞生长和虾青素生产的影响,并在70 L发酵罐上进行配比确定,在10 m3发酵罐上进行中试放大。结果表明:有利于细胞增殖的碳源为糖蜜、葡萄糖、麦芽糊精和麦芽糖浆,有利于类胡萝卜素合成的为麦芽糊精、麦芽糖浆、淀粉水解液和蔗糖,有利于虾青素合成的为麦芽糊精、淀粉水解液和麦芽糖浆;不同碳源组合中,糖蜜或葡萄糖与麦芽糊精组合时虾青素产量最高,分别达到(58.7±1.2)、(55.1±0.8)μg/m L;在70 L发酵罐中,葡萄糖和麦芽糊精质量比为1∶1时较佳,虾青素产量和干菌体虾青素含量分别为(341.0±7.2)μg/m L和4.08 mg/g(细胞干重);在10 m3发酵罐中进行工艺放大,虾青素产量和虾青素占类胡萝卜素比例分别为(360.3±0.8)μg/m L和42.2%,干菌体虾青素含量为3.88 mg/(g细胞干重),稍低于小试。该研究确定了法夫酵母虾青素生产的碳源种类和混合补加比例,为虾青素商业化生产提供了重要参考。     

红发夫酵母生产虾青素的培养基优化

从提高虾青素产量和降低生产成本综合考虑。研究选择了混合碳源、混合氮源、柠檬酸铵((NH_4)_3C_6H_5O_7)、Na_2HPO_4、K_2HPO_4、MgSO_4·7H_2O组成培养基。正交设计优选出的培养基为混合碳源(糖蜜40%、淀粉塘60%)30 g/L、混合氮源(玉米浆40%、(NH_4)_2SO_460%)7 g/L、MgSO_4·7H_2O 1.5 g/L、(NH_4)_3C_6H_5O_7 2g/L、Na_2HPO_4 2.0 g/L,用此优化培养基摇瓶培养红发夫酵母获得生物量16.92 g/L,虾青素含量903μg/g和虾青素产量15 279μg/L。     

不同时间流加番茄汁对红发夫酵母生产虾青素的影响

番茄红素作为虾青素合成的前体物质,适量添加对虾青素合成起到促进作用.由于成熟番茄汁中含有大量的番茄红素,可以作为番茄红素的替代品,而且番茄汁含有的其它营养物质对红发夫酵母的生长也有一定的积极作用.这样既促进了虾青素的合成,又节约了成本,故选择番茄汁作为流加物质.结果表明,培养36h时添加番茄汁,红发夫酵母72～120h生长状态平稳,且对提高虾青素含量和产量效果显著,培养96h虾青素含量高达1318 μg/g,虾青素产量也达到最高,为17.87mg/L.     

混合培养对雨生血球藻虾青素产量的影响

研究了不同光照强度及添加不同葡萄糖量进行混合培养对雨生血球藻虾青素产量的影响.研究表明,单位体积培养液虾青素产量因光照强度和葡萄糖添加量不同而不同,在光照强度为2500lx以及葡萄糖添加量为3g/L时,虾青素产量最高.光照强度和葡萄糖添加浓度对虾青素产量具有交互影响.通过中心组合试验,确定混合培养条件下达到最高虾青素产量所需要的葡萄糖添加量及光照强度分别为3.1616g/L和2605.66lx.此时的虾青素产量为41.06mg/L,是自养培养时的2.02倍.     

佐夫色绿藻高产虾青素的诱导条件及发酵工艺优化

本研究了不同诱导条件及光发酵罐培养工艺对混养佐夫色绿藻的影响,通过参数优化提高了藻细胞生物量和虾青素积累量.本研究首先系统地比较了在摇瓶系统中不同混合碳源和过氧化氢浓度对佐夫色绿藻的生长和虾青素积累的影响,并在光发酵罐中研究了恒定高光强、低光强-高光强以及低光强-高光强-补加过氧化氢三种不同发酵工艺对佐夫色绿藻积累虾青...     

利用控pH流加培养法提高雨生红球藻的生物量及虾青素产量

研究了一种新型培养方法——控pH流加培养法,可有效促进雨生红球藻生长并提高虾青素产量。测定雨生红球藻生长期间对营养盐的需求量,依据对数生长期藻液pH与硝酸盐和磷酸盐的含量的线性关系,确定流加液组成及流加方法。流加液的组成为7.88 g/L的KNO3,1.83 g/L的KH_2PO_4和24.0 g/L的CH_3COONa,监测对数生长期藻液pH变化情况,通过添加流加液将藻液的pH值持续控制在7.5~8.0内,雨生红球藻的细胞干重可达(1.42±0.04)g/L,虾青素产量可达(60.23±1.81)mg/L。     

红发夫酵母两步碳源流加培养

红发夫酵母是虾青素生产中最具潜力的生物来源。研究中应用PhaffiarhodozymaAs 2 15 5 7菌种 ,通过实验和分析方法确定碳源的两步流加培养方式 ,即第 1步用蔗糖和甘油的混合物 ;第 2步则流加蔗糖和乙醇的混合物。培养结果良好。     

红法夫酵母与环状芽孢杆菌混合培养破壁提取虾青素的研究

环状芽孢杆菌能以红法夫酵母细胞壁为唯一碳源进行发酵产胞壁溶解酶,将红法夫酵母与环状芽孢杆菌混合培养,可使环状芽孢杆菌在生长的同时产酶并逐渐溶解酵母细胞壁,从而提取虾青素。通过对红法夫酵母与环状芽孢杆菌两阶段混合培养条件的优化,总结出红法夫酵母产虾青素及酶法破壁提取的适宜工艺为:培养基初始糖浓度为30g/L,环状芽孢杆菌的最佳接种时间为红法夫酵母培养60～72h,接种量为10mL/L(1010个细胞/mL),接种后最适培养温度为30℃,pH为6.5,总发酵时间为120h,在此条件下,红法夫酵母虾青素产量可达到8960.2μg/L,虾青素最终提取率可达到96.8%。     

微生物法生产虾青素的研究进展

虾青素是一种萜烯类不饱和化合物,具有很强的抗氧化活性,同时还具有抗癌、增强免疫、护眼、保护心脑血管等其他诸多生理功能。天然虾青素主要来源于藻类、酵母以及海洋生物中。目前应用于市场的虾青素主要包括雨生红球藻虾青素、酵母虾青素以及化学合成虾青素。雨生红球藻的培养时间长,生长条件要求苛刻,使得虾青素价格昂贵;化学合成虾青素为混合构型虾青素,抗氧化能力低于天然虾青素,安全性较差。该文介绍了虾青素的结构性质、虾青素来源和生产方法,着重讲述红法夫酵母（Phaffia rhodozyma）生产虾青素的生物合成途径、发酵培养条件以及虾青素的破壁提取和纯化方法,为虾青素工业化生产提供一定理论基础。     

Effect of alternative C2 carbon sources on the growth, lipid, and γ-linolenic acid production of spirulina (Arthrospira platensis)

The influence of different organic carbon sources (glucose, ethanol, and acetic acid) at different concentrations (0.1, 0.5, and 1.0 g/L for batch and 1.0, 2.0, and 3.0 g/L for fed-batch) were studied in the mixotrophic production (using both light and carbon source) of γ-linolenic acid (GLnA) by spirulina (Arthrospira platensis). The obtained spirulina was analyzed in terms of biomass, lipid, and GLnA production. In the batch media, increasing the concentrations of glucose, ethanol, and acetic acid led to an increase in the biomass, lipid, and GLnA production. However, carbon sources at concentrations greater than 1.0 g/L in fed-batch media appeared to have no significant effects on the above parameters. It was also demonstrated that biomass, lipid, and GLnA production using ethanol and acetic acid could be as good as those achieved with the classic glucose-based culture media.     

Alcaligenes sp.NX-3产威兰胶的补料分批发酵工艺研究

在7.5L发酵罐上考察了Alcaligenes sp.NX-3产威兰胶的发酵工艺。选用葡萄糖为碳源,通过分析比较不同初糖浓度下的细胞比生长速率和产物比合成速率,进一步研究了不同补料方式对产胶的影响。结果表明,采用分批补糖发酵工艺,威兰胶产量较分批发酵提高了13.6%,而且有效地缩短了发酵周期。在50L发酵罐上进行补料分批发酵放大实验,威兰胶产量高达27.0g/L,葡萄糖转化率由初始的44%提高到54%。     

碱性中和与补料分批高密度培养保加利亚乳杆菌的研究

本实验采用葡萄糖反馈抑制补料、调节发酵液pH 值、碱性中和与补料分批复合培养等方法对保加利亚乳杆菌高密度培养进行研究。研究表明,碱性中和与补料分批复合培养条件为：保加利亚乳杆菌培养18h 和26h 时,向培养液补加碳氮比为0.67 的补料液5ml,并且每隔1.5h 调节pH 值至6.5,培养36h,培养液中保加利亚乳杆菌菌体浓度最高达9.02 × 1011CFU/ml,菌体干重达14.266g/L。     

Effect of ethanol on the production of carboxypeptidase Y using the GAL10 promoter in a Saccharomyces cerevisiae gal80 mutant

In the course of studying carboxypeptidase Y (CPY) production, we found that the expression level of the gene, which is under the control of the GAL10 promoter, increased in a Saccharomyces cerevisiae gal80 mutant grown in a medium containing ethanol as the sole carbon source. In the cultivation of the gal80 mutant KS58-2D/pCY303 carrying a multicopy plasmid, which contains the PRC1 gene fused to the GAL10 promoter, CPY production continued after the consumption of galactose. In this phase, the cells utilized ethanol as the carbon source. To increase the CPY production level, we examined the effect of carbon source feeding in a fed-batch culture. The production level in the fed-batch culture using ethanol was 1.3-fold higher than that in a batch culture and 1.6-fold higher than that in a fed-batch culture using galactose. By 5'-deletion analysis of the GAL10 promoter, the region between -256 and -232 was found to be important for the promoter activity in the gal80 mutant growing in the presence of ethanol.     

木醋杆菌流加静置培养生产细菌纤维素

木醋杆菌在传统的分批静置培养方式下产生的细菌纤维素具有良好的成膜性,但难于实现工业化生产,为了解决这个问题,对传统的静置培养装置进行改造,形成流加静置培养方式,能够将静置培养与动态培养的优点结合起来。对分批静置和流加静置2种培养方式进行比较,结果表明,流加培养时,细菌纤维素产量达到11.7g/L,其产量是分批培养的3.44倍,反应器的效率为0.585 g/(L·d),高于分批培养的0.425 g/(L·d)。流加培养条件下细菌纤维素产量和反应器效率得到提高,主要是由于在整个培养过程中营养供给充足,促进木醋杆菌的生长,使糖转化率及细菌平均比生长速率都达到较高水平,大量转化和合成细菌纤维素。     

Analysis of hemin effect on lactate reduction in Lactococcus lactis

Lactococcus lactis is a facultative anaerobic microorganism that produces lactate as the major product, and acetate and acetoin as by-products; some strains of this species produce an antimicrobial compound, nisin. Lactate has a strong inhibitory effect on L. lactis growth. On the other hand, hemin has a suppressive effect on lactate production during L. lactis growth under aerobic condition. To achieve the optimum effect of hemin on lactate amount reduction in L. lactis ATCC11454, cultures entailing various conditions were performed with and without hemin. In the culture with hemin, L. lactis growth and lactate reduction improved compared with those in the culture without hemin; that is, lactate production was suppressed by 1.8- and 1.3-fold under batch and fed-batch cultures, respectively. In microaerobic fed-batch culture with hemin, lactate production was sufficiently suppressed. This result suggests that microaerobic fed-batch culture could be applied to the maintenance of the low lactate amount. Under this condition, metabolic shift was observed from lactate to acetoin and acetate. However, no increase in nisin production was observed even though lactate production could significantly decrease in L. lactis ATCC11454.     

High butanol production by Clostridium saccharoperbutylacetonicum N1-4 in fed-batch culture with pH-Stat continuous butyric acid and glucose feeding method

A pH-stat fed-batch culture by feeding butyric acid and glucose has been studied in an acetone-butanol-ethanol (ABE) fermentation using Clostridium saccharoperbutylacetonicum N1-4. The specific butanol production rate increased from 0.10 g-butanol/g-cells/h with no feeding of butyric acid to 0.42 g-butanol/g-cells/h with 5.0 g/l butyric acid. The pH value in broth decreases with butyric acid production during acidogenesis, and then butyric acid reutilization and butanol production result in a pH increase during solventogensis. The pH-stat fed-batch culture was performed to maintain a constant pH and butyric acid concentration in the culture broth, but feeding only butyric acid could not support butyric acid utilization and butanol production. Subsequently, when a mixture of butyric acid and glucose was fed, butyric acid was utilized and butanol was produced. To investigate the effect of the feeding ratio of butyric acid to glucose (B/G ratio), several B/G ratio solutions were fed. The maximum butanol production was 16 g/l and the residual glucose concentration in broth was very low at a B/G ratio of 1.4. Moreover, yields of butanol in relation to cell mass and glucose utilization were 54% and 72% higher in pH-stat fed-batch culture with butyric acid than that of conventional batch culture, respectively.     

利用Crypthecodinium cohnii高密度发酵生产DHA的流加策略研究

利用流加策略实现了隐甲藻(Crypthecodinium cohnii)生产DHA的高密度发酵。根据隐甲藻间歇式发酵的特性,探讨最佳补料时间及流加液中最佳碳氮比,结果表明,在葡萄糖浓度降至4g/L左右时流加碳氮比为30∶1的营养液对隐甲藻的生长和脂肪酸积累最有利。根据以上结果,初始葡萄糖浓度25g/L,采用5d连续发酵多次流加策略,细胞干重达6.37%,DHA产量为4.08g/L。     

两阶段法培养红法夫酵母生产虾青素

在法夫酵母生长过程中,培养基组成是影响细胞生长和虾青素产量的重要因素。文中通过正交设计和响应面优化方法分别对红法夫酵母生长阶段培养基和虾青素合成阶段的培养基进行了优化研究。结果表明,碳源和氟源浓度对红法夫细胞的生长及虾青素的合成有明显影响,浓度为20 g/L的葡萄糖有利于细胞的生长;而50 g/L左右的葡萄糖和高C/N有利于虾青素的合成。在此基础上提出了两阶段发酵方案。经过两阶段发酵,红法夫酵母生物量和虾青素产量达到16.8 g/L和15.015 mg/L,分别比分批培养提高了56.3%和28.7%。