不同硝酸钾浓度对蔷薇藻生长及生理特性的影响

蔷薇藻(Rhodella reticulata)是属于红藻门的一种海洋单细胞微藻,其生长过程中产生藻胆蛋白、胞外多糖等生物活性物质.研究了不同硝酸钾浓度对蔷薇藻生长代谢的影响,分析测定蔷薇藻比生长速率、胞外多糖产量、藻蓝蛋白含量、色素含量、硝酸还原酶活性和SOD活性等参数.结果表明,与无氮培养基相比,蔷薇藻在以硝酸钾为氮源,浓度为0.75 g/L的条件下生长最好,硝酸还原酶活性最大,分别得到最大的藻蓝蛋白产量(24 mg/L)和类胡萝卜素含量(1.42 mg/L);当KNO3浓度为7.5 g/L时,获得最高叶绿素a含量为1.91 mg/L;高氮源有利于产糖,当KNO3浓度为30 g/L时,得到最高胞外多糖产量为1.633 g/L;SOD活性随硝酸钾浓度增加而增大.     

环境条件及摇瓶补糖策略对谷胱甘肽发酵的影响

研究了酵母摇瓶发酵中pH、装液量、初糖浓度、碳氮磷比和补糖方式对谷胱甘肽(GSH)发酵的影响。结果表明,GSH发酵适宜的初始pH和装液量分别为6 0和500ml锥形瓶内装液量60m1。初糖浓度对GSH发酵有较大的影响,超过12g／l,的初糖浓度将减少胞内GSH含量。应用计算得出的一种以控制比生长速率为目的的摇瓶补糖策略,在总糖浓度为26．2g／L下发酵12h,最终细胞浓度和胞内GSH含量分别达到8.78g／L和13．6mg／g,发酵液内GSH总量达到119．4mg／L,细胞对糖产率达到0.335g／g。     

龙须菜对重金属铜胁迫的生理响应

研究了大型海藻龙须菜(Gracilaria lemaneiformis)对不同浓度重金属铜(0、25、50、100、250和500 μg·L^-1)胁迫的生理响应.结果表明:当Cu²⁺浓度≥50 μg·L^-1时,龙须菜藻体的相对生长速率显著下降,最大光化学量子产量、最大相对电子传递速率和相对电子传递效率呈相同的变化趋势.随着Cu²⁺浓度的升高,龙须菜藻体最大净光合速率和光饱和点显著降低,而光补偿点显著升高,叶绿素a、类胡萝卜素和藻胆蛋白含量则呈先升高后下降的趋势;当Cu²⁺浓度达到500 μg·L^-1时,叶绿素a、类胡萝卜素和藻胆蛋白含量显著下降.说明龙须菜在低浓度Cu²⁺胁迫下具有一定的抵抗能力,而当Cu²⁺浓度≥50 μg·L^-1时,会对藻体生理活动造成显著的抑制作用.     

微鞘藻胞外多糖在沙漠土壤成土中的作用

研究了沙坡头藻结皮中的优势物种——微鞘藻的胞外多糖对沙漠表层土壤水分分布状况的影响及其光合特性。随着胞外多糖浓度的升高 ,土壤持水量增大 ,土壤水分蒸发速率降低 ,水分在土壤中运动的速率被延缓。藻总水溶性多糖产率为 2 8.82 % ,胞外水溶性多糖产率为 1 6 .0 9% ;多糖产量在稳定期生物量最大时增长最快。微鞘藻光合作用最适温度为 2 5℃ ,最适光强为 40 0 μE.m- 2 .s- 1,在 0 .3 mol/ L Na Cl浓度下其光合作用活性被强烈抑制 ;微鞘藻的吸收光谱表明它比生长在湿润地区的藻类含有更多的胡萝卜素成分 ;叶绿素 a荧光 (Fv/ Fm)分析表明在失水约为初始湿重的 5 6 %时 Fv/ Fm值恢复到水培养的 96 % ;其光合作用特性有利于对逆境的适应和生物量的积累。表明微鞘藻胞外多糖可影响沙漠表层土壤水分分布状况 ,并可作为一种潜在的生物肥料 ,在荒漠原始成土过程中扮演着重要角色。     

极小曲丝藻生长与光合活性对硅浓度的响应

为探究南水北调中线干渠硅藻快速增殖与硅浓度间的内在关系, 选取干渠全年优势种极小曲丝藻(Achnanthidium minutissimum)为研究对象, 研究不同初始硅浓度(0、2、4、6和8 mg·L−1)对极小曲丝藻生长和光合生理特性的影响。结果显示, 不同浓度硅对极小曲丝藻比生长速率具有显著影响(P<0.05)。硅浓度0 mg·L−1时, 细胞密度、比生长速率、叶绿素a浓度、类胡萝卜素浓度和光合系统Ⅱ(PSⅡ)最大光化学效率(Fv/Fm)相对于添加硅的试验组, 均显著降低(P<0.05), 未出现明显的叶绿素荧光诱导曲线(OJIP)特征, 检测不到光合放氧。随硅浓度升高(28 mg·L−1), 藻细胞密度、比生长速率和最大现存生物量均明显上升; 硅浓度2 mg·L−1处理组的细胞叶绿素a和类胡萝卜素浓度均显著低于另外3个硅处理组(P<0.05)。硅浓度4 mg·L−1、6 mg·L−1和8 mg·L−1处理组的细胞叶绿素a和类胡萝卜素浓度、Fv/Fm和光合放氧速率均无显著差异。研究表明硅是硅藻快速增殖的限制因素; 中线干渠内硅浓度较高(约5 mg·L−1), 满足硅藻大量增殖需要, 是导致硅藻快速增殖、占据优势的重要因子。     

影响深红酵母和白地霉产生胞外蛋白和多肽的因素

采用白地霉2.498-47在以蛋白胨为氮源发酵时,只在开始pH3.2左右产生胞外蛋白,NH_4~+则无此限制,还有助于胞外多肽的产生。深红酵母2.280胞外蛋白的最大产率约在pH5.8;尿素作唯一氮源效果最好,与蛋白胨合并使用,可产胞外蛋白约800μg/ml。酵母膏对生长和产胞外蛋白很必要,不能用玉米浆和麦芽汁代替。用0.05N NaOH浸洗不同pH和氮源所培养的细胞都能浸出胞内蛋白,接近400μg/ml。发酵产生胞外蛋白自一日后开始,至二日接近最大量,以后逐渐减少。多肽产生起自第二日,五日未达到最大量。两株酵母菌都不表现蛋白酶活性,因此这个现象还不能解释。     

1株结皮真菌胞外多糖的初步研究

探究一株真菌胞外多糖最佳生产时间及其组成成分。通过对其发酵液胞外多糖含量及上清液黏度的测定,确定其最佳产糖时间,并用此条件提取胞外多糖粗品,用蒽酮-比色法测得提取的胞外多糖粗品的糖含量,薄层色谱法对所得的胞外多糖进行组分分析。当发酵至第5天时该株真菌的生物量、胞外多糖产量及发酵液的黏度均达到最高峰,分别为0.606 4 g/60 mL、20.22μg/mL和4.74 mPa/s,并且胞外多糖含量与发酵上清液黏度在一定程度上呈正相关。用蒽酮-比色法测得提取的胞外多糖粗品的糖含量为19.58μg/mL,并用4 mol/L的硫酸完全水解,硅胶G薄层层析,甲醇-氯仿(1:1,体积比)展层,苯胺-二苯胺-磷酸显色,表明其组成成分可能为D-甘露糖、D-半乳糖和D-葡萄糖。该菌株所产的胞外多糖具有提高结皮能力和防沙保水作用,为利用微生物治沙提供参考和种质资源。     

营养盐因子对细基江蓠繁枝变种氮、磷吸收速率的影响

在实验室条件下,研究了营养盐因子对细基江蓠繁枝变种的氮、磷吸收速率的影响。分别进行了营养盐浓度与温度双因子试验,氮磷比与荧 浓度双因子试验肽不同化合态氮比例单因子试验。（1）氮、磷的吸收速率随营养盐浓度的升高而增大,氮的吸收速率在21℃,总氮浓度为100μmol/L时最大,达2.58μmol/(g.h);磷的吸收速率在31℃,总磷为6．3μmol/L时最大,达0.17μmol/(g.h),温度与营养盐浓度有显著的交互作用效应。（2）当氮浓度一定时,环境氮磷比对氮的吸收率无显著影响,但对磷的吸收速率有显著影响,藻中收的氮磷比随环境氮磷比的不同而变化,（3）对3种不同化合态氮的吸收速率与培养液中各种氮占总无机氮的比例呈正相关,当三比例相同时,对NH4^ -N、NO3^--N格NO2^-析吸收分别占总吸收氮的40．7％、28．5％和30．8％。     

粗柄羊肚菌菌丝体多糖及胞外多糖的提取工艺

为了研究粗柄羊肚菌菌丝体多糖及胞外多糖的最佳提取工艺条件,采用超声波提取菌丝体多糖、菌丝体发酵液浓缩提取胞外多糖的方法。通过单因素试验和L9(34)正交试验设计,探讨二者的最佳提取工艺条件。研究结果表明菌丝体多糖的最佳提取工艺条件:料液比1∶20(g∶m L),超声波处理温度70℃,超声时间20 min,超声提取次数2次;胞外多糖的最佳提取工艺条件:浓缩倍数1∶5,浓缩温度50℃,醇沉浓度95%,p H为6。该项工艺研究得到菌丝体粗多糖平均含量56.761 2 mg/g,胞外多糖平均含量1.275 4 mg/m L,多糖产量稳定,且此试验方法稳定可行。     

短期连续剪切对光生物反应器内海带配子体细胞生长及其恢复能力的影响

以不同搅拌速率(0~1000r/min)对海带(Laminaria japonica)配子体细胞施加短期(0~60h)连续剪切,卸载剪切力之后细胞静止恢复23.5d,在此期间研究不同搅拌速率分别在连续剪切和恢复期间对细胞生长及其恢复能力的影响。研究结果表明,连续剪切期间,90r/min下细胞叶绿素浓度积累达到最大值2.36mg/L;中高速搅拌速率(270~1000r/min)下叶绿素浓度迅速下降,胞内氮磷池释放,1000r/min下细胞损伤率为静止对照样的18倍。恢复期间,所有组别细胞均呈现较强的恢复能力。此外,在连续剪切实验中,海带配子体细胞内叶绿素浓度比干重更能有效的表达生物量浓度,除细胞损伤率外,磷源释放可以作为判别细胞受损的参考指标之一。     

Effects of nitrogen,phosphorus, iron and silicon on growth of five species of marine benthic diatoms

The effects of nitrogen, phosphorus, iron and silicon on growth of five species of marine benthic diatoms, namely Navicula patrickae, Nitzschia panduriformis, Navicula thienemannii, Nitzschia longissima and Navicula atomus were studied by single factor experiments and the optimal concentration ratios of the four nutrient elements beneficial for diatoms growth were screened out separately using the L₉ (3⁴) orthogonal design. The results highlighted that nitrogen, phosphorus, iron and silicon all had highly significant effects on growth of five diatoms while the diatoms growth rates reached the highest averagely in the 2nd to the 6th culture day. In addition, the optimal concentrations (mg/L) of four nutrients suitable for diatoms growth were found higher than that in f/2 medium except that Nitzschia longissima had the same concentration of nitrogen as that in f/2 medium which is optimal for growth. Moreover, the optimal growth concentrations of four elements for five diatoms varied in the range of 12.36–74.16 mg/L for nitrogen, 1.70–3.98 mg/L for phosphorus, 2.00–4.00 mg/L for iron, 23.01–69.03 mg/L for silicon, respectively. By means of the orthogonal test of four nutrients for five benthic diatoms, the optimal concentration ratios N:P:Fe:Si (mg/L) were obtained as follows: 74.16:2.27:3.33:23.01 for N. patrickae; 37.08:3.98:4.00:11.50 for N. panduriformis; 49.44:3.98:3.33:34.51 for N. thienemannii; 12.36:1.70:4.00:11.50 for N. longissima; 74.16:2.27:4.00:69.03 for N. atomus.     

Influence of nutritional conditions on the mycelial growth and exopolysaccharide production in Paecilomyces sinclairii

AIMS: The objective of the study was to optimize the submerged culture conditions for the production of exopolysaccharide from Paecilomyces sinclairii. METHODS AND RESULTS: The optimal temperature and initial pH for exopolysaccharide production by Paecilomyces sinclairii in shake flask culture were found to be 30 degrees C and 6.0, respectively. Sucrose (60 g l(-1)) and corn steep powder (10 g l(-1)) were the most suitable carbon and nitrogen source for exopolysaccharide production. CONCLUSIONS: Under optimal culture medium, the maximum exopolysaccharide concentration in a 5-l stirred-tank fermenter indicated 7.4 g l(-1), which was approximately three times higher than that in basal medium. The maximum specific growth rates (micro max) and yield coefficient (Y(P/S)) in the optimal culture medium was 0.16 h(-1) and 0.19, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The optimal culture conditions reported in this article can be widely applied to the processes for submerged cultures of other mushrooms.     

Optimization of submerged culture requirements for the production of mycelial growth and exopolysaccharide by Cordyceps jiangxiensis JXPJ 0109

AIMS: The objective of the present study was to investigate the optimal culture requirements for mycelial growth and exopolysaccharide production by Cordyceps jiangxiensis JXPJ 0109 in submerged culture. METHODS AND RESULTS: The effects of medium ingredients (i.e. carbon and nitrogen sources, and growth factor) and other culture requirements (i.e. initial pH, temperature, etc.) on the production of mycelia and exopolysaccharide were observed using a one-factor-at-a-time method. More suitable culture requirements for mycelial growth and exopolysaccharide production were proved to be maltose, glycerol, tryptone, soya bean steep powder, yeast extract, medium capacity 200 ml in a 500-ml flask, agitation rate 180 rev min(-1), seed age 4-8 days, inoculum size 2.5-7.5% (v/v), etc. The optimal temperatures and initial pHs for mycelial growth and exopolysaccharide production were at 26 degrees C and pH 5 and at 28 degrees C and pH 7, respectively, and corresponding optimal culture age were observed to be 8 and 10 days respectively. According to the primary results of the one-factor-at-a-time experiments, the optimal medium for the mycelial growth and exopolysaccharide production were obtained using an orthogonal layout method to optimize further. Herein the effects of medium ingredients on the mycelial growth of C. jiangxiensis JXPJ 0109 were in the order of yeast extract > tryptone > maltose > CaCl2 > glycerol > MgSO4 > KH2PO4 and the optimal concentration of each composition was 15 g maltose (food-grade), 10 g glycerol, 10 g tryptone, 10 g yeast extract, 1 g KH2PO4, 0.2 g MgSO4, and 0.5 g CaCl2 in 1 l of distilled water, while the order of effects of those components on exopolysaccharide production was yeast extract > maltose > tryptone > glycerol > KH2PO4 > CaCl2 > MgSO4, corresponding to the optimal concentration of medium was as follows: 20 g maltose (food-grade), 8 g glycerol, 5 g tryptone, 10 g yeast extract, 1 g KH2PO4, and 0.5 g CaCl2 in 1 l of distilled water. CONCLUSIONS: Under the optimal culture requirements, the maximum exopolysaccharide production reached 3.5 g l(-1) after 10 days of fermentation, while the maximum production of mycelial growth achieved 14.5 g l(-1) after 8 days of fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the submerged culture requirements for mycelial growth and exopolysaccharide in C. jiangxiensis, and this two-step optimization strategy in this study can be widely applied to other microbial fermentation processes.     

Formation of carotenoids by rhodotorula glutinis in whey ultrafiltrate

The growth and carotenoid biosynthesis of the yeast Rhodotorula glutinis was studied by cocultivation with Lactobacillus helveticus in cheese ultrafiltrate containing 3.9% and 7.1% lactose. By growing this mixed culture in a 15-L fermentor MBR AG (Switzerland) at an air flow rate of 0.5 L/L min and agitation at 220 rpm for 6 days, a total yield of carotenoids of 268 mug/g dry cells wasobtained. Carotenoids were formed almost parallel with the cell growth, anda maximum production was reached at an early stationary phase. A high-performance liquid chromatographic system (HPLC) permitting simultaneous determination of major carotenoid pigments was used. The three main pigments (torularhodin, beta-carotene, and torulene) were formed in Rhodotorula glutinis, and reached a maximum concentration as follows: 182.0, 43.9, 23.0 mug,g dry cells. (c) 1994 John Wiley & Sons, Inc.     

Exopolysaccharide production and mycelial growth in an air-lift bioreactor using Fomitopsis pinicola

For effective exopolysaccharide production and mycelial growth by a liquid culture of Fomitopsis pinicola in an air-lift bioreactor, the culture temperature, pH, carbon source, nitrogen source, and mineral source were initially investigated in a flask. The optimal temperature and pH for mycelial growth and exopolysaccharide production were 25degrees C and 6.0, respectively. Among the various carbon sources tested, glucose was found to be the most suitable carbon source. In particular, the maximum mycelial growth and exopolysaccharide production were achieved in 4% glucose. The best nitrogen sources were yeast extract and malt extract. The optimal concentrations of yeast extract and malt extract were 0.5 and 0.1%, respectively. K2HPO4 and MgSO4 x 7H2O were found to be the best mineral sources for mycelial growth and exopolysaccharide production. In order to investigate the effect of aeration on mycelial growth and exopolysaccharide production in an air-lift bioreactor, various aerations were tested for 8 days. The maximum mycelial growth and exopolysaccharide production were 7.9 g/l and 2.6 g/l, respectively, at 1.5 vvm of aeration. In addition, a batch culture in an air-lift bioreactor was carried out for 11 days under the optimal conditions. The maximum mycelial growth was 10.4 g/l, which was approximately 1.7-fold higher than that of basal medium. The exopolysaccharide production was increased with increased culture time. The maximum concentration of exopolysaccharide was 4.4 g/l, which was about 3.3-fold higher than that of basal medium. These results indicate that exopolysaccharide production increased in parallel with the growth of mycelium, and also show that product formation is associated with mycelial growth. The developed model in an air-lift bioreactor showed good agreement with experimental data and simulated results on mycelial growth and exopolysaccharide production in the culture of F pinicola.     

磷浓度对小环藻、大型溞和金鱼藻三者相互作用的影响

为了解磷浓度对水生植被恢复和生物操纵效果的影响, 分别用小环藻(Cyclotella sp.)、大型溞(Daphnia magna)和金鱼藻(Ceratophyllum demersum)代表浮游植物、浮游动物和大型沉水植物建立水生微宇宙模型, 在25℃、2600 lx光强和11 mg/L氮浓度条件下, 分别研究小环藻与大型溞、小环藻与金鱼藻、小环藻-大型溞-金鱼藻共培养时4种磷浓度(0.05、0.1、0.5和2 mg/L)下小环藻、大型溞、金鱼藻的增长率以及培养液中氮磷去除率的变化。结果表明: 小环藻与大型溞、小环藻与金鱼藻两两共培养时, 磷浓度为0.05-2 mg/L时, 金鱼藻和大型溞均生长良好, 小环藻受到明显抑制, 其密度保持较小幅度的正增长。在小环藻-大型溞-金鱼藻三者共培养时, 在0.05-2 mg/L的磷浓度范围内大型溞和金鱼藻生长良好, 与两两共培养相比, 小环藻则受到了更大程度的抑制, 在磷浓度为0.05-0.1 mg/L时藻密度呈现负增长. 这说明在水生态系统中, 大型浮游动物和沉水植物对浮游藻类的联合控制效果远好于各自单独的控制效果, 该控制效果随磷浓度的提高而减弱, 以0.1 mg/L的磷浓度为最佳。在实验结束后测定氮磷去除率发现, 在最低磷浓度(0.05 mg/L),即磷限制时, 水中磷去除率最高, 在最高磷浓度(2 mg/L), 即氮限制时, 水中氮去除率最高。     

Optimization for growth of Rhodocyclus gelatinosus in seafood processing effluents

Tuna condensate was a better substrate than shrimp-blanching water or effluent from a frozen-seafood plant for growing Rhodocyclus gelatinosus under anaerobic conditions in the light. One strain out of four examined, R7, gave the highest biomass (4.0 g/l), cell yield (0.32 g cell/g COD), and COD removal (78%) in 1:10 (v/v) diluted tuna condensate. Shrimp-blanching water added to the tuna condensate further increased growth rate, biomass and COD removal. Optimal growth was at pH 7.0 and 3000 Lux light intensity. Acetate, pyruvate, glucose, glutamate, propionate or malate added to the tuna condensate did not increase cell yield, carotenoid or bacteriochlorophyll content or biomass protein. A maximum cell mass of 5.6 g/l (containing 50% protein) and 86% COD removal were obtained after 5 days' incubation under optimal conditions.     

格木幼苗对硝普钠-酸铝互作的生理响应

以格木（Erythrophleum fordii Oliv.）幼苗为材料,采用双因素完全随机设计实验方法,测定不同处理幼苗的光合色素和可溶性糖等生理指标,研究格木幼苗对硝普钠（SNP）-氯化铝（AlCl₃）互作的生理响应。结果显示,格木幼苗叶片中叶绿素a、叶绿素b和类胡萝卜素含量均在处理4（0.2 mmol/L AlCl₃、0.1 mmol/L SNP）时最高,在处理9（0.8 mmol/L AlCl₃、0 mmol/L SNP）时含量最低,而叶片中丙二醛（MDA）、游离脯氨酸含量则相反;叶片可溶性糖、可溶性蛋白含量在处理4时最高,在处理9时最低;处理10（0.8 mmol/L AlCl₃、0.1 mmol/L SNP）的超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性最高。施加SNP后,格木幼苗叶片中的叶绿素a、叶绿素b、类胡萝卜素、可溶性糖、可溶性蛋白含量及SOD、POD、CAT活性均显著高于未施加SNP处理。相关性分析表明,叶绿素a、类胡萝卜素、总叶绿素和可溶性蛋白含量等指标间均呈极显著正相关（P < 0.01）。本研究结果得出,低浓度AlCl₃（0.2 mmol/L）胁迫可促进格木幼苗的生长,添加外源SNP对高浓度AlCl₃（0.8 mmol/L）胁迫格木幼苗产生的毒害具有一定的缓解作用,可在格木幼苗的培育及抗性研究中推广应用。     

Production of β-carotene by a mutant of Rhodotorula glutinis

Wild strains of Rhodotorula glutinis and R. rubra were investigated concerning their carotenoid production, proportion of beta-carotene and cell mass yield. R. glutinis NCIM 3353 produced 2.2 mg carotenoid/l in 72 h; and the amount of beta-carotene was 14% (w/w) of the total carotenoid content (17 microg/g cell dry weight). It was subjected to mutagenesis using UV radiation for strain improvement. Out of 2,051 isolates screened, the yellow coloured mutant 32 produced 120-fold more beta-carotene (2,048 microg/g cell dry weight) than the parent culture in 36 h, which was 82% (w/w) of the total carotenoid content. Mutant 32 was grown on different carbon and nitrogen sources. The best yield of beta-carotene (33+/-3 mg/l) was obtained when glucose and yeast extract were supplied as carbon and nitrogen sources, respectively. Divalent cation salts further increased the total carotenoid content (66+/-2 mg/l) with beta-carotene as the major component (55+/-2%, w/w).     

Improvement of the conversion of polystyrene to polyhydroxyalkanoate through the manipulation of the microbial aspect of the process: a nitrogen feeding strategy for bacterial cells in a stirred tank reactor

Pseudomonas putida CA-3 has been shown to accumulate the biodegradable plastic polyhydroxyalkanoate (PHA) when fed styrene or polystyrene pyrolysis oil as the sole carbon and energy source under nitrogen limiting growth conditions (67 mg nitrogen per litre at time 0). Batch fermentation of P. putida CA-3 grown on styrene or polystyrene pyrolysis oil in a stirred tank reactor yields PHA at 30% of the cell dry weight (CDW). The feeding of nitrogen at a rate of 1mg N/l/h resulted in a 1.1-fold increase in the percentage of CDW accumulated as PHA. An increase in the rate of nitrogen feeding up to 1.5mg N/l/h resulted in further increases in the percentage of the cell dry weight composed of PHA. However, feeding rates of 1.75 and 2mg N/l/h resulted in dramatic decreases in the percentage of cell dry weight composed of PHA. Interestingly nitrogen was not detectable in the growth medium after 16 h, in any of the growth conditions tested. A higher cell density was observed in cells supplied with nitrogen and thus further increases in the overall production of PHA were observed through nitrogen feeding. The highest yield of PHA was 0.28 g PHA per g styrene supplied with a nitrogen feeding rate of 1.5mg/l/h.