通气量对灵芝菌丝体液态深层发酵合成灵芝三萜的影响

采用5 L搅拌式发酵罐研究灵芝菌丝体液态深层发酵合成灵芝三萜的扩大培养条件,考察了不同条件下的通气量对灵芝菌丝体生长、比生长速率、灵芝三萜合成、比合成速率、还原糖消耗、还原糖比消耗速率、铵根离子消耗以及铵根离子比消耗速率的影响。研究结果表明,通气量能够显著影响菌丝体的生长以及灵芝三萜的合成,当通气量为8 L/min时,菌丝体合成灵芝三萜得率和生产强度最高,分别为0.204 g/L和0.00131 g/(L·h),最大菌丝体得率达到8.77 g/L。发酵过程中菌丝体最大比生长速率为0.0704 1/h,灵芝三萜最大比合成速率为0.0256 1/h,还原糖和铵根离子最大比消耗速率分别为0.562 1/h和0.0171 1/h。     

碳源种类及碳氮比对眼点拟微绿球藻生长密度、油脂含量和脂肪酸组成的影响

研究了三种碳源Na2CO3、NaHCO3、葡萄糖对眼点拟微绿球藻生长密度和油旨含量的影响,实验结果表明相对于葡萄糖,无机碳源NaHCO3更利于眼点拟微绿球藻的生长.以NaHCO3为碳源,研究了在不同的接种密度、NaNO3浓度下,C/N对眼点拟微绿球藻生长密度和油脂含量的影响.实验结果表明,C/N对眼点拟微绿球藻生长密度的影响与接种密度和NaNO3浓度有关,在高的NaNO3浓度时,C/N对眼点拟微绿球藻生长密度的影响很小;在低的NaNO3浓度时,随着C/N比的增加,微绿球藻的生长密度先增加后下降,存在最佳的C/N比.最佳的C/N比随接种密度而变化,在接种密度为OD440=0.10时,最佳C/N比为3,当接种密度提高到OD440=0.70时,最佳C/N比增加到5.NaNO3浓度和C/N对微藻油脂含量均有较大影响,在不同的接种密度和NaNO3浓度下都表现为C/N=1时最利于微藻油脂的积累,这与卡尔文循环过程中核酮糖-1,5-二磷酸羧化酶/加氧酶的活性有关.本实验的最佳产油培养条件为以NaHCO3为碳源,初始接种密度为OD440=0.70,C/N=1∶1,CNaNO3=0.225g/L,此时油脂产率为56.7 mg/(L·d),EPA产率为6.5 mg/(L·d).     

斯达氏油脂酵母利用混合糖发酵产油脂

研究了斯达氏油脂酵母Lipomyces starkeyi2#利用葡萄糖-木糖混合糖为碳源生长和油脂积累特性。L.star-keyi2#利用70 g/L葡萄糖和70 g/L木糖作为碳源在30℃下摇瓶发酵96 h,糖利用率均达90%以上,菌体生物量分别为14.1 g/L和13.1 g/L,油脂质量分数分别为55.7%和52.6%。相同条件下该菌株利用混合糖(葡萄糖46 g/L,木糖24 g/L)为碳源时总糖利用率、生物量和油脂质量分数分别为75.1%,15.0 g/L和40.0%。借助于P lackett-Burm an设计法和单因子实验法对培养条件进行了优化,结果表明发酵96 h混合糖利用率可达到97.3%,发酵120 h后混合糖利用率、生物量和菌体油脂质量分数分别达99.5%、19.0 g/L和52.6%。生物量得率和油脂得率分别达到27%和14%。     

螺旋藻连续培养与动力学模型

在内循环气升式光生物反应器中 ,研究了钝顶螺旋藻 (SpirulinaplatensisGeitler)细胞的连续生长及其对碳源底物的利用特性。结果表明 :随着稀释率的增大 ,反应器中碳源浓度和细胞浓度分别呈上升和下降趋势 ,它们之间的关系可用Monod类型的方程很好地加以关联。细胞产率和碳消耗速率与稀释率的关系存在峰值现象 :在本实验条件下 ,最大细胞产率为 0 .36 2g/(L·d) ,最大碳消耗速率为 0 .177g/(L·d) ,此时稀释率为 0 .45 /d ,细胞浓度为OD560 =1.2 82 ,细胞对碳的得率系数为 2 .0 5 0g/g。所提出的连续培养动力学模型与实验数据拟合较好     

螺旋藻批式与连续培养及其生长动力学

在内循环气升式光生物反应器中,分别研究了螺旋藻细胞在批式和连续培养条件下的生长特性,结果表明：Richards模型和指数衰减模型可较好地描述批式培养时细胞和碳源底物浓度与培养时间的关系;批式培养时最大细胞生长速率为0371g/d/L,细胞对碳的得率系数为3.439g/gC;连续培养时随着稀释率的增大,细胞和底物浓度分别呈下降和上升趋势;连续培养时最大细胞产率为0.362g/L/d,最佳稀释率为0.45/d,细胞对碳的得率系数为2.050g/gC;所提出的连续培养动力学模型可较好地拟合实验数据。     

甲醇抑制时重组毕赤酵母发酵的特性研究

本文对毕赤酵母进行了恒化培养研究。以甲醇为唯一碳源时,在稀释率较低时(D<0.048 h^-1),连续培养系统操作很稳定。但在稀释率高时(D>0.048h^-1),连续培养系统的定态点不止一个,实验不能维持,故采用比生长速率恒定的分批流加培养进行研究。结果表明,毕赤酵母的生长符合Andrew普遍化底物抑制模型。综合考虑水蛭素的生成、底物的消耗,在生产中维持甲醇浓度为限制性浓度(0.5 g/L),且维持比生长速率为0.02 h^-1时,水蛭素Hir65的比生成速率达到最大值0.2 mg/(g·h)且甲醇的比消耗速率为0.04 g/(g·h)。     

耐高糖米根霉菌株的生理特性

高糖发酵是提高产物浓度的常用方法,但天然菌株不能耐受高的糖浓度。为了解析高糖环境对菌株生理代谢的影响,以驯化获得的耐高糖菌株和原始菌株为对象,研究不同糖浓度对二者生理特性的影响。结果发现:与原始菌株相比,耐高糖菌株细胞膜不饱和脂肪酸的含量、胞内三磷酸腺苷(ATP)含量较高,且表现出更强的侧系呼吸强度。在此基础上,尝试通过调节碳氮比(C/N)或添加甘氨酸的方式补足原始菌株的代谢不足。结果表明:在150 g/L糖质量浓度条件下,调节C/N为500时,原始菌株富马酸产量可由37.4 g/L增至40.2 g/L。而添加0.5g/L甘氨酸,原始菌株富马酸产量可以增至43.8 g/L,提高了17.1%。     

重组巴氏毕赤酵母恒化培养动力学及代谢迁移特性研究

通过对甲醇营养型毕赤酵母基因工程菌以碳源甘油为限制性基质进行恒化培养动力学试验 ,结果认为 :(1 )细胞光密度与其干、湿重呈线性关系 ,当细胞光密度 (OD60 0 )为 1 0 0时细胞湿重 (WCW)为 1 2 8 3g L ,细胞干重 (WDW)则为 2 2 9g L ;(2 )基因工程菌P .pastoris的生长与限制性基质甘油残留浓度的关系符合Monod关系式 ,通过 1 μ对 1 S进行线性回归得 μmax=0 .366h- 1,Ks=0 .1 82 3g L ,经参数推导甘油最大菌体得率系数YG =0 .54g g ,菌体维持生长消耗底物系数m =0 .0 0 69g (g·h) ;氧最大菌体系数YX O2 =30 .96g moL ,菌体维持生长时消耗氧系数mO2 =0 .0 0 0 8mol (g·h) ,最适理论稀释速率Dm =0 .341h- 1;(3)从氨水的消耗速率和呼吸商 (RQ)的变化认为随着比生长速率 (μ)的增大 ,甘油代谢流从糖原异生和磷酸戊糖途径线性地向糖酵解和三羧酸循环途径进行代谢迁移 ,即糖酵解和三羧酸循环途径的代谢流量在线性地增大     

重要理化因子对小球藻生长和油脂产量的影响

本文采用通气培养的方法研究了N、P、Fe3 、盐度、光照强度、温度对小球藻(Chlorella sp. XQ-200419)生长速率、生物量和油脂产量的影响。主要结果如下：N浓度对小球藻的生长和油脂产量均有显著的影响,在KNO3浓度0.05—0.3g/L范围内,小球藻生长速率随N浓度的增加而提高,并积累更多的生物量,而油脂含量随之递减,KNO3浓度为0.3g/L时,油脂产量最高。小球藻对P浓度变化的适应范围很大,K2HPO4浓度在10—160mg/L范围内,对小球藻的生长和油脂产量都没有显著影响。在小球藻培养后期补加不同浓度Fe3 对其生长速率没有显著影响,总脂含量随着Fe3 浓度升高呈现上升的趋势,均比对照有极显著提高,Fe3 浓度为0.75mmol/L时油脂产量最高。盐度对小球藻的生长有一定的抑制作用;油脂含量先随着盐度的增大而提高,当NaCl浓度达到0.6mol/L, 油脂含量又显著降低;油脂含量和油脂产量均在盐度为0.2mol/L时最高。光照强度对处于生长后期的小球藻的生长影响不大,但影响其油脂积累,小球藻的油脂含量和产量随光照强度的增大而显著提高,当光照强度增至280μmolm-2s-1时,油脂含量和油脂产量最高。温度对小球藻的生长速率、生物量、油脂含量和油脂产量都有显著的影响,在15-40℃范围内,随着培养温度的升高,生长速率、生物量、油脂含量和油脂产量都经历了一个先上升然后下降的过程,适合小球藻生长、积累油脂的温度范围是20-35℃,30-35℃时油脂产量最高,40℃时生物量、油脂含量和产量都最低。理化因子对生长和油脂含量的影响分为两种情况：1. 温度、光强、铁浓度和盐度的影响表现为在适宜生长的条件下提高油脂含量,这种模式可以称为“适宜模式”;2. 氮浓度的影响表现为在不利于生长的条件下提高油脂含量,这种模式可以称为“胁迫模式”。两种模式都可以提高油脂含量,但是,只有适宜模式才可以提高油脂产量。在筛选小球藻优良产油藻种时要注意,只有在适宜的培养条件下油脂含量高的藻种才具有高产油潜力。     

圆红冬孢酵母利用生物乙醇废水-木薯粉水解液发酵产油

【目的】获得能够高效降解生物乙醇废水化学需氧量(COD)的圆红冬孢酵母菌株,评估废水初始COD浓度对驯化菌株生长的影响,将木薯粉生产微生物油脂和高浓度有机废水降解过程整合,以生物乙醇废水为水源制备生物乙醇废水-木薯粉水解液培养基,明确产油效率高、生物乙醇废水COD降解率高的初始还原糖浓度。【方法】采用在高浓度的生物乙醇废水中进行多次驯化的方法,获得能够适应废水环境的圆红冬孢酵母菌株;采用双酶水解法对加入乙醇废水中的木薯粉进行水解;采用重量法监测生物量浓度变化,采用酸热法提取油脂,重铬酸钾法监测COD,DNS法测定废水还原糖浓度,凯氏定氮法测定总氮,钼酸铵比色法测定总磷。【结果】通过驯化筛选得到一株能耐受高浓度生物乙醇废水的优势菌株Rhodosporidium toruloides D5。以未稀释的废水为培养基,驯化菌株的最终生物量浓度和COD降解率分别为3.8 g/L和75.0%。采用生物乙醇废水-木薯粉水解液发酵时,控制初始还原糖浓度低于30 g/L时,生物量浓度和油脂浓度随初始还原糖浓度的升高而升高,均在120 h时达到最高COD降解率,初始还原糖浓度对达到的最大COD降解率无明显影响,废水N、P去除率分别达到99%和92%以上。【结论】在未经稀释的高浓度生物乙醇废水中可获得较高的生物量浓度;采用高浓度生物乙醇废水-木薯粉水解液培养基发酵产油,初始还原糖浓度为30 g/L,可在保证高油脂产量的同时,实现废水COD的高效降解,有效回收利用废水中残余的N、P源,从而降低微生物油脂生产和废水处理成本,研究结果可为开发廉价微生物油脂生产技术提供有用的参考。     

Studies on the removal of dyes from a synthetic textile effluent using barley husk in static-batch mode and in a continuous flow,packed-bed,reactor

Yarrowia lipolytica LGAM S(7)1 presented remarkable growth on industrial glycerol used as sole carbon substrate. Nitrogen-limited flask cultures were accompanied by restricted synthesis of reserve lipid, whilst amounts of citric acid were produced extracellularly. On the contrary, high amounts of reserve lipid (up to 3.5 g/l, 43% w/w of lipids in dry biomass) were produced in highly aerated continuous cultures. Lipid production was favoured at low specific dilution rates whilst fat-free material yield increased over the whole range of D (h(-1)). The maximum volumetric productivity obtained was 0.12 g lipid/1 h. Storage lipid composition did not present remarkable changes in the specific dilution rates tested. Oleate and linoleate were the dominant cellular fatty acids.     

Continuous ethanol production from Jerusalem artichoke tubers. I. Use of free cells of Kluyveromyces marxianus

The Continuous fermentation of Jerusalem artichoke juice to ethanol by free cells of Kluyveromyces marxianus UCD (FST) 55-82 has been studied in a continuous-stirred-tank bioreactor at 35 degrees C and pH 4.6. A maximum yield of 90% of the theoretical was obtained at a dilution rate of 0.05 h(-1). About 95% of the sugars were utilized at dilution rates lower than 0.15 h(-1). Volumetric ethanol productivity and volumetric biomass productivity reached maximum values of 7 g ETOH/L/h and 0.6 g dry wt/L/h, respectively, at a dilution rate of 0.2 h(-1). The maintenance energy coefficient for K. marxianus culture was found to be 0.46 g sugar/g biomass/h/ Oscillatory behavior was following a change in dilution rate from a previous steady state and from batch to continuous culture. Values of specific ethanol production rate and specific sugar uptake were found to increase almost linearly with the increase of the dilution rate. The maximum specific ethanol production rate and maximum specific sugar uptake rate were found to be 2.6 g ethanol/g/ cell/h and 7.9 sugars/g cell/h, respectively. Washout occurred at a dilution rate of 0.41 h(-1).     

Lipid accumulation in an oleaginous yeast (Candida 107) growing on glucose in single-stage continuous culture.

Lipid accumulation of Candida 107, grown at dilution rates from 0.03 to the maximum of 0.21/h, with carbon, nitrogen, phosphate, and magnesium limitations in a chemostat, was maximal at about 40% (wt/wt) with nitrogen-limited medium at a dilution rate of 0.06/h, giving an efficiency of substrate conversion of 22 g of lipid per g of glucose consumed. At higher dilution rates the lipid content decreased. With carbon-limited growth, the highest lipid content (14%, wt/wt) was at the maximum dilution rate. High lipid contents also occurred with phosphate + nitrogen as double limitations of growth, with the lipid content of the yeast (about 35%, wt/wt) continuing to be near maximum at dilution rates also near maximum (0.17/h), thus giving the highest specific rate of lipid formation of any growth conditions (0.59 g of lipid/g of yeast per h). However, the efficiency of substrate utilization was only 5.2 g of lipid formed per 100 g of glucose consumed. The composition of the fatty acyl residues within the lipid remained constant over many weeks if the steady-state conditions remained unchanged. With carbon-limited growth, the degree of unsaturation of the fatty acids markedly decreased as the dilution rate was increased, but with nitrogen limitation the reverse trend was seen. In all cases, linoleic and oleic acids were the principal fatty acyl residues affected, and their relative proportions always varied in opposite directions. When magnesium was a limiting nutrient, there was a considerable increase in the proportion of myristic acid produced within the lipid. Neutral lipids (predominantly triglycerides) varied from 66 to 92% of the total lipid from carbon- and nitrogen-limited growth; phospholipids (varying from 2 to 25%) were highest in nitrogen-limited growth. The fatty acyl residues within each lipid fraction showed the same variations with changing growth rates.     

Kinetics of growth and fatty acid production of Rhodotorula glutinis

Summary The effect of culture conditions, especially nitrogen/carbon (N/C) ratio and temperature, on acyl lipid profiles in the oleaginous yeast Rhodotorula glutinis NRRL Y 1091 have been investigated. Cultivation of the microorganism under N-limited conditions (N/C < 0.14 g/g) resulted in enhanced fatty acid (FA) cell content but a reduced relative amount of polyunsaturated fatty acids (PUFA). The maximal FA productivity was obtained for a 0.025 N/C ratio resulting from the arrangement between the specific rate of FA synthesis and the concentration of lipid-free biomass. Under nitrogen-non-limiting conditions, cells grown at lower temperatures had a higher PUFA content and the maximal productivity of -linolenic acid was obtained by shifting the temperature of the culture from 30° C to 25° C. Offprint requests to: A. Pareilleux     

Candida zeylanoides as a new yeast model for lipid metabolism studies: effect of nitrogen sources on fatty acid accumulation

Lipid homeostasis is well-known in oleaginous yeasts, but there are few non-oleaginous yeast models apart from Saccharomyces cerevisiae. We are proposing the non-oleaginous yeast Candida zeylanoides QU 33 as model. The aim of this study was to investigate the influence of the carbon/nitrogen ratio and the type of nitrogen source upon oil accumulation by this yeast grown on shake flask cultures. The maximum biomass was obtained in yeast extract (2.39?±?0.19 g/l), followed by peptone (2.24?±?0.05 g/l), while the highest content of microbial oil (0.35?±?0.01 g/l) and the maximum lipid yield (15.63 %) were achieved with peptone. Oleic acid was the predominant cellular fatty acid in all culture media (>32.23 %), followed by linoleic (>15.79 %) and palmitic acids (>13.47 %). The highest lipid yield using glucose and peptone was obtained at the C/N ratio of 200:1.     

Efficient lipid production with Trichosporon fermentans and its use for biodiesel preparation

Effects of medium components and culture conditions on biomass and lipid production of Trichosporon fermentans were studied. The optimal nitrogen source, carbon source and C/N molar ratio were peptone, glucose and 163, respectively. The favorable initial pH of the medium and temperature were 6.5 and 25 degrees C. Under the optimized conditions, a biomass of 28.1 g/l and a lipid content of 62.4% could be achieved after culture for 7 days, which were much higher than the original values (19.4 g/l and 50.8%) and the results reported by other groups. T. fermentans could grow well in pretreated waste molasses and a lipid yield of 12.8 g/l could be achieved with waste molasses of 15% total sugar concentration (w/v) at pH 6.0, representing the best result with oleaginous microorganisms on agro-industrial residues. Addition of various sugars to the pretreated molasses could efficiently enhance the accumulation of lipid and the lipid content reached as high as above 50%. Similar to vegetable oils, the lipid mainly contains palmitic acid, stearic acid, oleic acid and linoleic acid and the unsaturated fatty acids amount to about 64% of the total fatty acids. The microbial oil with an acid value of 5.6 mg KOH/g was transesterified to biodiesel by base catalysis after removal of free fatty acids and a high methyl ester yield of 92% was obtained.     

Influence of cultivation conditions on lipid production by Cryptococcus albidus

Summary Cryptococcus albidus var. albidus CBS 4517 was able to accumulate lipid under nitrogen-limited as well as excess-nitrogen conditions. The highest lipid-producting capacity was, however, observed in nitrogen-limited cultivations. In nitrogen-limited batch cultures, a lipid content of 34% (w/w) in biomass and a maximum specific lipid productivity of 37 mg lipid/g lipid-free biomass·h, was determined. The yield of lipid from glucose was about 0.15 g/g in nitrogen-limited and 0.11 g/g in excess-nitrogen cultures.In a nitrogen-limited fed-batch culture, 12.4 g/l lipid was produced at 90 h of cultivation and the cells contained 46.3% (w/w) lipid.Higher lipid yield and cellular lipid content were observed when inorganic nitrogen sources were used compared with organic. The choice of carbon source was seen to influence growth as well as lipid production and the highest yields of lipid were obtained when glucose, maltose or mannitol was used.A cultivation temperature of 20°C provided the highest lipid productivity compared to 25°C and 30°C. Addition of citrate to the growth medium was seen to have a stimulating effect on the specific lipid productivity.     

Optimization of pineapple pulp residue hydrolysis for lipid production by Rhodotorula glutinis TISTR5159 using as biodiesel feedstock

The higher lipid productivity of Rhodotorula glutinis TISTR5159 was achieved by optimizing the pineapple pulp hydrolysis for releasing the high sugars content. The sequential simplex method operated by varied; solid-to-liquid ratio, sulfuric acid concentration, temperature, and hydrolysis time were successfully applied and the highest sugar content (83.2 g/L) evaluated at a solid-to-liquid ratio of 1:10.8, 3.2% sulfuric acid, 105 °C for 13.9 min. Moreover, the (NH₄)₂SO₄ supplement enhanced the lipid productivity and gave the maximum yields of biomass and lipid of 15.2 g/L and 9.15 g/L (60.2%), respectively. The C16 and C18 fatty acids were found as main components included oleic acid (55.8%), palmitic acid (16.6%), linoleic acid (11.9%), and stearic acid (7.8%). These results present the possibility to convert the sugars in pineapple pulp hydrolysate to lipids. The fatty acid profile was also similar to vegetable oils. Thus, it could be used as potential feedstock for biodiesel production.     

Enhanced microbial lipid production by <Emphasis Type="Italic">Cryptococcus albidus</Emphasis> in the high-cell-density continuous cultivation with membrane cell recycling and two-stage nutrient limitation

As a potential feedstock for biofuel production, a high-cell-density continuous culture for the lipid production by Cryptococcus albidus was investigated in this study. The influences of dilution rates in the single-stage continuous cultures were explored first. To reach a high-cell-density culture, a single-stage continuous culture coupled with a membrane cell recycling system was carried out at a constant dilution rate of 0.36/h with varied bleeding ratios. The maximum lipid productivity of 0.69 g/L/h was achieved with the highest bleeding ratio of 0.4. To reach a better lipid yield and content, a two-stage continuous cultivation was performed by adjusting the C/N ratio in two different stages. Finally, a lipid yield of 0.32 g/g and lipid content of 56.4% were obtained. This two-stage continuous cultivation, which provided a higher lipid production performance, shows a great potential for an industrial-scale biotechnological production of microbial lipids and biofuel production.     

Lipid productivity and fatty acid composition in Chlorella and Scenepdesmus strains grown in nitrogen-stressed conditions

Thirty Chlorella and 30 Scenedesmus strains grown in nitrogen-stressed conditions (70 mg L^?1 N) were analyzed for biomass accumulation, lipid productivity, protein, and fatty acid (FA) composition. Scenedesmus strains produced more biomass (4.02?±?0.73 g L^?1) after 14 days in culture compared to Chlorella strains (2.57?±?0.12 g L^?1). Protein content decreased and lipid content increased from days 8 to 14 with an increase in triacylglycerol (TAG) accumulation in most strains. By day 14, Scenedesmus strains generally had higher lipid productivity (53.5?±?3.7 mg lipid L^?1 day^?1) than Chlorella strains (35.1?±?2.8 mg lipid L^?1 day^?1) with the lipids consisting mainly of C16–18 TAGs. Scenedesmus strains generally had a more suitable FA profile with higher amounts of saturated fatty acids and monounsaturated fatty acids (MUFAs) and a smaller polyunsaturated fatty acid (PUFA) component. Chlorella strains had a larger PUFA component and smaller MUFA component. The general trend in the FA composition of Chlorella strains was oleic > palmitic > α-linolenic = linoleic > eicosenoic > heptadecenoic > stearic acid. For Scenedesmus strains, the general trend was oleic > palmitic > linoleic > α-linolenic > stearic > eicosenoic > palmitoleic > heptadecenoic acid. The most promising strains with the highest lipid productivity and most suitable FA profiles were Scenedesmus sp. MACC 401, Scenedesmus soli MACC 721, and Scenedesmus ecornis MACC 714. Although Chlorella sp. MACC 519 had lower lipid productivity, the FA profile was good with a lower PUFA component compared to the other Chlorella strains analyzed and a low linolenic acid concentration.