紫球藻的光生物反应器培养

利用光生物反应器就碳源及其供给形式、光强和反应器的操作条件等影响紫球藻生长的因素进行了研究.结果表明:反应器良好的液体循环速度有助于培养液中每个细胞对光能的吸收利用,使紫球藻细胞可以获得较大的生长速度;利用CO2为碳源可以获得比利用NaHCO3更大的生长速度,有效地提高了紫球藻的生物量产量.在本实验条件下,细胞的生长速度和生物量产率分别达到了0.952 d-1和42.31 g/(m2.d).     

Enhanced Production of Podophyllotoxin by Fed‐batch Cultivation of Podophyllum hexandrum

Podophyllum hexandrum (Indian May Apple) was successfully cultivated in a 3 L stirred tank bioreactor under low shear conditions in batch and fed‐batch modes of operation. A statistically optimized culture medium was used for the batch cultivation of Podophyllum hexandrum. Under optimum culture conditions of P. hexandrum, the batch culture showed a growth‐associated product formation with a maximum biomass of 21.4 g/L dry cell weight (DCW) basis and a podophyllotoxin production of 13.8 mg/L in 26 d. A mathematical model was developed to design the nutrient feeding strategies for a fed‐batch cultivation to prolong the productive log phase of cultivation. The fed‐batch cultivation was able to enhance the biomass and podophyllotoxin accumulation to 48 g/L (DCW basis) and 43.2 mg/L, respectively, in 60 d. The volumetric productivity of podophyllotoxin in fed‐batch cultivation was found to be 0.72 mg/(L. d) as opposed to 0.53 mg/(L. d) in batch cultivation under optimized culture conditions.     

用于微藻培养的气升式光生物反应器

基于微藻光自养培养特性 ,构建了具有较大面积体积比的 15 L内外光源相结合的气升式光生物反应器 ,考察了两种不同形态藻细胞培养体系中 ,光强随细胞浓度及光程距离衰减的规律 ,得到了描述光衰减的数学关系式 ,即在鱼腥藻 712 0培养体系中为 I=I0 exp[- (0 .0 131+0 .987OD750 )·L],在聚球藻 70 0 2培养体系中为 I=I0 exp[- (- 0 .0 2 39+0 .0 777OD750 )·L],并据此对培养过程中光强沿反应器径向的动态分布情况进行了估算。在该反应器中进行了鱼腥藻 712 0和聚球藻 70 0 2两种蓝藻的光自养培养 ,藻细胞培养终密度分别达到 1.5 3g/ L和 3.4g/ L ,体积产率分别为 0 .31g L-1d-1和 0 .5 7g L-1d-1,说明该反应器适合于微藻的高密度培养     

有效碱度对螺旋式自循环厌氧反应器性能的影响

采用模拟废水研究了有效碱度对螺旋式自循环(SPAC))厌氧反应器运行性能的影响.结果表明,SPAC厌氧反应器的最大容积负荷为53.0 g/(L·d),化学耗氧量(CDD)去除率为84.2%,容积负荷承载力为44.6 g/(L·d),容积沼气产率为21.50 L/(L·d).在最佳工况下,碱度供给速率为9.6 g/(L·d).碱度消耗速率为4.8 g/(L·d),有效碱度为1 000 mg/L.进水碱度与进水COD之比为0.19,出水碱度与挥发性有机酸(VFA)之比(ALKout/VFA)为1.75,消耗碱度与去除COD之比为0.10.     

豆粕水解液为氮源细菌厌氧流加发酵生产L-乳酸

采用细菌厌氧发酵法生产L-乳酸,由实验确定了最佳接种量、发酵温度和pH调节剂,考察了初始葡萄糖浓度对L-乳酸生产的影响,确定初始糖浓度为70~90 g/L时得率、产率、最终生物量分别达到92.68 g/g, 3.17 g/(L×h)和8.5′107 mL-1. 为进一步降低L-乳酸生产成本,以豆粕水解液为氮源代替酵母粉,同时应用流加发酵技术,L-乳酸产量、得率、产率及转化率分别达到155 g/L, 95.5 g/g, 1.64 g/(L×h)和96.9%. 在保证L-乳酸最终浓度的同时可降低生产成本,为进一步工业化奠定了基础.     

发状念珠藻细胞液体悬浮培养的研究

为解决天然发状念珠藻生长缓慢的问题,从天然藻体中分离出细胞对其进行液体悬浮培养。结果表明,液体悬浮培养可以显著提高发状念珠藻细胞的生长速度,同时提高发状念珠藻胞外多糖的产量。采用BG-11培养基,添加2.5 g/L硝酸钠,培养温度25℃,光照强度40μmol/(m2.s),转速130 r/min,培养20 d细胞干质量浓度达到1.05 g/L,胞外多糖产量89.9 mg/L。培养发状念珠藻细胞的基本成分和细胞培养液的HPLC指纹图谱分析表明其生化组成和多糖的合成与天然发状念珠藻相似。与其他人工培养方法相比,液体悬浮培养更适合大规模工业化生产。     

重组大肠杆菌产胆固醇氧化酶的指数流加策略

以指数流加策略高密度培养重组大肠杆菌,发酵生产胆固醇氧化酶。通过对比生长速率的分阶段控制使得细胞干重达40.128g/L。确定了最佳的诱导时机为菌体对数生长期的中期,产酶速率为1287.31U/(L·h)。菌体产酶水平达6436.56U/L,生产强度为459.75U/(L·h),实现了胆固醇氧化酶的高效生产。     

铜离子与葡萄糖协同补料对球头三型孢菌产赤藓糖醇的影响

为考察葡萄糖和铜离子盐协同补料发酵对球头三型孢菌产赤藓糖醇的影响,在5L发酵罐中先采用不同浓度的葡萄糖进行分批发酵,然后采用优化的葡萄糖浓度并添加CuSO₄·5H₂O进行发酵研究。结果表明,初始葡萄糖浓度为300g/L的赤藓糖醇产量最大为44.52g/L,其体积生产速率为0.371g/（L·h）、转化率为0.167g/g。在此浓度葡萄糖的基础上添加30mg/L的CuSO₄·5H₂O后,赤藓糖醇产量达到49.62g/L,提高了11.5%。进一步控制总糖浓度为300g/L,且初始浓度为200g/L,分别进行单独补糖和协同补糖与铜离子的补料发酵,结果赤藓糖醇产量分别为47.25g/L和55.31g/L,比初始300g/L的葡萄糖分批发酵分别提高了6.1%和24.2%。特别地,协同补糖与CuSO₄·5H₂O后,赤藓糖还原酶（erythrose reductase,ER）的活性在84h达到最大,为0.152U/mg,比单独补糖时提高了18.8%;通过铜离子盐和葡萄糖的协同补料发酵可显著提高赤藓糖醇的产量,最终使赤藓糖醇产率达到0.461g/（L·h）。     

Multistage continuous cultivation of blue-green alga spirulina maxima in the flat tank photobioreactors with recycle

Spirulina maxima was continuously cultivated in four 64-liter flat tank photobioreactors in cascade, under continuous 30 klx fluorescent light and nonaseptic condition, in an industrial grade synthetic medium which bubbled with atmospheric air. The increase in the number of stages resulted in the higher biomass concentration and allowed operation with a dilution rate D higher than the alga specific growth rate μ. However, multistage operation does not permit a significant increase in productivity. The recycling of the all culture medium with biomass increased the net productivity of the first two stages of the multistage system. The flat tank design offered great advantages over other geometrical configurations by occupying less space for indoor cultivation, minimizing the water loss by evaporation, and permitting a greater flexibility in construction, installation, and operation. This system was operating continuously for two years in our laboratory for indoor mass production of S. maxima biomass. The flat tank configuration gave the highest areal productivity Pa = 60.5 g/m²·d and highest volumetric productivity Pv = 1.17 g/L·d when operated as a single stage continuous cultivation with high solid recycle. These values are the highest ever reported for any indoor or outdoor mass production system for S. maxima.     

洁霉素废水厌氧生物处理研究

采用上流式厌氧污泥床反应器,在34℃下处理洁霉素废水。通过进水稀释与不稀释的实验对比,分析探讨了有毒有害物质、碱度、容积负荷等对厌氧生物处理的影响。洁霉素废水的CODCr质量浓度为16.8~24.3g/L,BOD5质量浓度为7.2~9.9g/L。在进水不稀释条件下,CODCr负荷为5.7kg/(m3·d),进水CODCr的质量浓度为16.8g/L,进水BOD的质量浓度为7.2g/L时,CODCr平均去除率为72%,BOD5平均去除率为84%。     

藏红花胚性愈伤组织的发生及其调控

为提高藏红花胚性愈伤组织的繁殖系数和出芽率,促进其生长和分化,以建立藏红花离体快繁体系,解决藏红花资源短缺问题,研究了藏红花胚性愈伤组织的发生及其调控. 结果表明,获得的藏红花球茎1细胞系具有良好的胚性愈伤组织发生能力. 胚性愈伤组织生长的优化条件为：在添加3.0 mg/L 6-BA, 0.25 mg/L NAA和400 mg/L CH的B5固体培养基上,22℃下全时暗培养25 d,繁殖系数达到9 g/g. 胚性愈伤组织出芽的优化条件为：在添加3.0 mg/L 6-BA, 0.25 mg/L NAA和400 mg/L CH的1/2 B5固体培养基上,在22℃及光照强度31.74 mmol/(m2×s)条件下,每天光照10 h,暗培养14 h,培养45 d出芽率达到44.7%,高于国外报道的20%.     

高产量、高分子量透明质酸发酵条件优化

研究了搅拌转速、初糖浓度及通气量对兽疫链球菌Streptococcus zooepidemicus WSH24发酵生产透明质酸的影响. 研究结果表明,搅拌转速对透明质酸产量及分子量影响很大,搅拌转速为200 r/min时透明质酸产量达到5.3 g/L,平均分子量达到1.88×106 Da,产率系数为0.13 g/g;初始葡萄糖浓度为65.8 g/L时有利于透明质酸的生产,产量达5.9 g/L,平均分子量达1.90×106 Da,产率系数为0.17 g/g;通气量对透明质酸的发酵也有较大影响,通气量为1.2 L/(min·L)时透明质酸的产量及分子量均高于0.5 L/(min·L)时的发酵结果.     

室内外培养对耐温微藻Desmodesmus sp. F51细胞生长和叶黄素积累的影响

在室内培养耐温微藻F51,考察了培养温度、自来水及非灭菌培养基对细胞生长及叶黄素积累的影响,并与50 L柱状光生物反应器户外分批培养情况进行对比. 结果表明,藻株在35℃下的生物质产率和叶黄素产率最大,分别为767.5和3.05 mg/(L?d). 低温培养有利于叶黄素积累,但生长速率减缓. 用自来水及非灭菌培养基培养时,藻株无明显的生长抑制及染菌现象,在13.0?27.6℃户外培养温度下,藻株最高生物质产率、二氧化碳固定率和叶黄素产率分别为236.3, 444.2和1.17 mg/(L?d).     

利用Clostridium saccharobutylicum DSM 13864 连续发酵生产丁醇

为使用廉价糖质原料进行连续发酵生产丁醇提供理论依据和实验指导,以Clostridium saccharobutylicum DSM 13864为发酵菌种,考察了稀释率和温度等因素对以葡萄糖为原料的四级连续发酵生产丁醇的影响。结果表明：高稀释率有利于酸的积累和菌体的生长,低稀释率有利于溶剂的生产。当稀释率为0.05 h?1,4个发酵罐温度控制在37 ℃时,总溶剂产量为11.57 g/L,其中丁醇7.29 g/L,生产率为0.145 g/(L?h)。在0.05 h?1稀释率的条件下进行变温连续发酵证实低温有利于溶剂的积累,当一级和二级罐温度控制在37 ℃,三级和四级罐温度控制在33 ℃时总溶剂产量最高为13.69 g/L,其中丁醇为8.36 g/L,生产率为0.171 g/(L?h)。     

Biomass production by a thermotolerant yeast: Hansenula polymorpha

Biomass production at high temperature by Hansenula polymorpha as part of a lignocellulosic utilizing process was studied. Compromise growth conditions (45°C and pH = 4.8) with an eventual saccharification step were established. The effects of stirring rate and initial glucose concentration on biomass yield coefficient, volumetric productivity and maximal cell density were determined. Process optimization led to a fed-batch fermentation process: high yield (0.63 g dry cell g^?1 glucose), volumetric productivity (1.3 g dry cell dm^?3 h^?1) and cell concentration (60 g dry cell dm^?3) were obtained. At these conditions, significant arabitol excretion (18 g dm^?3) as a unique by-product associated with cell mass production was obtained, making more interesting a high temperature operating process.     

Continuous ethanol production from wood hydrolysate by chemostat and total cell retention culture

Chemostat and total cell retention cultures with internal filter system ofSaecharomyc.es cerevisiae H1-7 were carried out to produce ethanol from wood hydrolysate. Maximum ethanol productivity obtained in a chemostat with the aeration rate of 1 vvm was 3.79 g/(L·h). This was 20% higher than that in a chemostat without aeration. However, the substrate was not completely consumed at the dilution rate with the maximum productivity. The realistic productivity, which has higher than 99% conversion rate of substrate, was. 2.95 g/(L·h). The maximum productivity in the total cell retention culture was 6.65 g/(L·h) at the dilution rate of 0.19 h¹ and the residual glucose concentration was negligible.     

高硅高铁含铜渣氧压酸浸过程

对火法炼铅产高硅高铁含铜渣进行了氧压硫酸浸出过程研究. 结果表明,浸出时间、氧分压、酸量、浸出温度和搅拌速度对Cu的浸出率和浸出液中Fe含量有显著影响,溶液初始含Cu2+量对Cu浸出率影响不明显. 最佳工艺条件为：时间2 h,氧分压0.8 MPa,硫酸浓度46.6 g/L,温度(150±2)℃,搅拌速度600 r/min,复合絮凝剂A用量30~100 g/m3. 在该条件下,Cu浸出率>95%,浸出液中Fe<5 g/L,硫转化率20%~30%,料浆过滤速度约0.8 m3/(m2×h).     

新型气升式反应器中浸没/非浸没周期对水母雪莲细胞培养的影响

应用新型周期浸没气升式反应器，进行了水母雪莲细胞的浸没悬浮和非浸没静置交替循环培养，研究了培养过程中的关键条件?浸没/非浸没周期对水母雪莲细胞生长、黄酮合成及细胞聚集体调控的影响，发现5 min/4 h为最佳的循环周期. 在此循环周期下，当通气量为40 L/h、接种量(干重DW)为2~3 g/L时，经20 d的培养，细胞聚集体分布最佳, 细胞干重达9.6 g/L，黄酮类化合物的含量及黄酮类化合物的产量分别可达35.2 mg/g (DW)及338 mg/L.     

转hTNF-α基因聚球藻培养条件的初步研究

以转hTNF-α基因聚球藻7002为对象,在摇瓶培养下对其生长条件进行了初步研究. 结果表明,当光照强度为100 μmol/(m2×s)时,藻细胞生长速率最大;35oC为其最佳培养温度;氯化钠浓度为12~24 g/L生长良好,最适浓度为24 g/L;转hTNF-α基因的聚球藻7002能利用铵盐和硝酸盐为氮源,其中硝酸盐对其生长最有利,硝酸钠最适浓度为1 g/L;加入有机碳源可以显著促进藻细胞生长,其中5 g/L的蔗糖对其促进作用最明显.     

紫球藻培养条件优化

对影响紫球藻生长的KH2PO4,N4NO3,N3HCO3,VBI和VB12等五种主要营养因素进行了优化,获得了以海水为基础的优化培养基配方,有效地提高了紫球藻的生物量产量,为该微藻的进一步高密度放大培养提供了依据．此外,研究发现紫球藻也能利用有机氮源,在培养中流加适量的尿素能促进藻体细胞的生长．