Streptoverticillium mobaraense分批发酵生产谷氨酰胺转胺酶:氨基酸代谢流分析及其初始淀粉浓度的影响

对Streptoverticilliummobaraense分批发酵合成谷氨酰胺转胺酶 (MTG)的氨基酸代谢流分布进行了理论分析 ;通过考察MTG分批发酵过程中的一些参数的变化情况 ,包括菌体干重 (DCW )、MTG酶活及残糖 (RSC)、菌体产率系数、MTG产率系数以及MTG的生产强度等 ,确定了适宜的初始淀粉浓度 ;对MTG发酵过程中氨基酸的代谢流分布进行了计算并作了较详细的分析 .研究结果表明 ,初始淀粉浓度 30g/L较适宜 ,DCW最高达 2 0 .9g/L ,酶活最高可达 2 .8UmL-1,菌体产率系数、MTG产率系数和MTG的生产强度分别为 0 .94g/g、12 5 .4U/g和 5 1.9UL-1h-1.氨基酸代谢流分布表明 ,只要发酵液中游离氨基酸充分 ,则细胞生长和产物形成就会活跃 ,限制细胞生长和产物的一个重要因素可能是MTG对氨基酸氮源的交联行为 ,表明可以采用不同氮源和控制氮源水平的方法改进菌体生长和MTG合成 .图 9表 2参 9     

混合培养微生物利用甘油补料发酵生产乙醇研究

采用浸麻芽孢杆菌和红曲菌990691用甘油混合发酵生产乙醇.结果表明,分批发酵中高浓度的甘油对乙醇发酵有着较强的抑制作用,分批发酵最佳甘油浓度为0.217 mol L-1.在分批发酵的基础上补料发酵,考察了不同甘油浓度的补料液和不同补料时间对乙醇发酵的影响.最终确定乙醇补料发酵较优的工艺条件为:反应器1 L,装液量700 mL红曲发酵液,甘油初始浓度为0.217 mol L-1,以补料方式每隔60 h分5次补加0.217 mol L-1甘油浓度的红曲发酵液,每次补加100 mL,发酵培养360 h.当乙醇最高浓度达0.221 mol L-1,乙醇总产率0.628 mmol h-1,乙醇/甘油转化率达87%(mol mol-1).与分批发酵相比,补料发酵很大程度解除了高浓度甘油的抑制作用,有效地利用了甘油,提高了乙醇的产量,且乙醇产率较为稳定.     

吸附菌HX5对活性艳蓝KN-R的吸附脱色作用

研究了吸附菌HX5对活性艳蓝KNR的吸附脱色作用,碳源、氮源、盐度和染料浓度对KNR吸附脱色的影响,以及HX5生长菌体对KNR的脱色机理.结果表明,菌株HX5对KNR脱色的最佳碳、氮源分别为葡萄糖和硫酸铵;碳源浓度在10g/L以上时,可使200mg/L的KNR完全脱色,碳源浓度过高,脱色效果不显著;HX5对KNR脱色的最佳氮源浓度为0.75g/L,在0～2%的浓度范围内,盐度对脱色无显著影响;染料对菌株HX5具有一定的生长抑制毒性,但对于400mg/L的KNR,脱色率仍可达95.1%;HX5生长菌体对KNR作用96h内主要为生物吸附作用,96h外则可能发生了生物降解.图5表2参9     

Clostridium papyrosolvens厌氧发酵产氢研究

采用分批培养研究了从高浓度厌氧产氢活性污泥中筛选的优势菌种Clostridium papyrosolvens的发酵产氢能力.结果表明:该菌有较强的高糖耐受性和耐酸性,当葡萄糖浓度为30 g/L、pH阶段性控制在4.5时,发酵44 h葡萄糖消耗率为83.7%,总产气量达到3 081.3 mL/L,最高产气率为187.5 mL L-1 h-1,氢气含量为67.5%,比产氢率达1.06 mol(H2)/mol(葡萄糖).研究中选用了廉价的发酵产氢培养基,以玉米浆为氮源,以还原铁粉作氧化还原电位控制剂,省去了牛肉膏、蛋白胨等昂贵氮源以及L-半胱氨酸、维生素、无机离子等高成本组分,显著降低了纯菌发酵的培养基成本,获得了较好的产氢效果.图5表2参23     

温度对赤霉素发酵的影响及优化策略

温度是影响赤霉素发酵的重要因素,研究温度与菌丝生长和次级代谢物合成的关系将有助于发酵工艺优化.采用5 L全自动发酵罐,调控罐温在25-35℃范围内进行赤霉素发酵,分时记录菌体浓度、残糖浓度和产物赤霉素浓度变化情况.发酵过程数据经微分处理后,比较分析在不同温度下赤霉素发酵过程中菌比生长速率、菌得率及赤霉素比合成速率的变化特征.结果显示:最适菌丝生长温度是32℃,其比生长速率和得率分别为0.571/h和0.431 g/g;适宜GA3合成的温度是28℃,最大比合成速率为2.161 mg g~(-1) h~(-1).进而提出赤霉素发酵过程变温控制轨迹:发酵0-18 h,控温28℃;18-40 h,温度32℃;40-60 h控制温度为30℃;60 h后,控制温度28℃.采用此温度控制策略进行GA3发酵,赤霉素终浓度达到2 294 mg/L,比恒温28℃发酵浓度提高了11.14%.本研究表明赤霉素发酵过程分阶段变温控制策略能提高产量,具有产业应用前景.     

Optimization of fermentation process for saccharide-based fuel ethanol production by Saccharomyces cerevisiae北大核心CSCD

低成本、高产量的发酵工艺是实现工业燃料乙醇经济和环境可持续性发展的关键,而不需要重大基础设施改变或投资.为获得酿酒酵母(Saccharomyces cerevisiae)利用甘蔗汁生产燃料乙醇的最优发酵工艺,首先对发酵体系的氮源条件进行优化;其次,在单因素试验基础上,以乙醇发酵效率为响应值,通过响应面法优化了燃料乙醇生产的发酵工艺,并通过补料分批发酵技术在5 L发酵罐中进一步扩大发酵.结果表明,以1.0 g/L (NH)SO和1.0 g/L酵母提取物作为发酵氮源,乙醇发酵效率和得率比对照可分别提高4.80%、9.52%.响应面设计获得的最优发酵工艺条件为在总糖浓度150.0 g/L、酵母提取物浓度2.0 g/L、发酵时间24.5 h、pH5.0、外加(NH)SO浓度1.0 g/L时,最高乙醇发酵效率可达到91.10%.在5 L发酵罐中采用补料分批发酵获得的最终乙醇浓度达到98.92 g/L,发酵效率维持在90%左右,乙醇生产力最高达到3.81 g Lh.本研究获得了一种高效生产糖质燃料乙醇的发酵工艺,可在较短时间内获得高浓度乙醇且消耗较少氮源,结果可为进一步利用糖质原料进行高效生物炼制及高浓度乙醇工业化生产提供参考.(图6表6参30)     

葡萄糖和氯化钠对米根霉利用鱼粉废水生成乳酸的影响

米根霉AS3.254能较好地利用鱼粉废水生成乳酸,葡萄糖和NaCl浓度能显著影响该过程的实现.当外加葡萄糖浓度为30g/L,发酵培养72h的鱼粉废水CODCr去除率为97.8%,乳酸产量为19.4g/L,生物量为3.56g/L;最大乳酸得率和总氮去除率分别为0.66g/g和55%.鱼粉废水中较高浓度氯化钠(NaCl)能抑制菌株生长,降低其产酸能力.当NaCl浓度大于12g/L时,菌株生长被完全抑制,产酸能力完全丧失.图4表1参20     

均匀设计法优化樟芝产三萜液体发酵条件

为在兼顾生物量的基础上提高液体发酵中三萜的百分含量,首先通过单因子试验筛选出最佳的碳源、氮源、无机盐,然后采用均匀设计(UD)对其培养基配比和培养条件进行优化试验,建立适当的数学模型,并对模型进行验证确定优化条件.单因子试验结果表明玉米淀粉既能促进菌丝体的生长,又能促进三萜的合成;麸皮虽然对菌体生长略有不利,但是对三萜的积累作用特别显著;硫酸镁对菌丝体生长效果不显著,但是对三萜的合成比较有利.均匀设计试验结果表明,菌丝体干重最大的培养条件和三萜的最优培养条件相差甚远.菌丝体干重的最优培养条件为:1 L培养液中玉米淀粉47 g,麸皮47 g,硫酸镁0.5 g,初始pH 3.0,培养7 d;三萜最佳培养基配方为:1 L培养液中玉米淀粉20 g,麸皮20 g,硫酸镁1.85 g,初始pH 3.0,培养16 d.菌丝体干重在最优条件下能达到15.58(±0.37)g L-1;三萜百分含量在最优培养条件下高达6.04(±0.03)%,三萜的百分含量比基础发酵结果和报道的最高发酵结果3.18%提高了90%,因此,均匀设计法能有效优化液体发酵培养条件     

氮源对枯草芽孢杆菌WSHDZ-01合成过氧化氢酶的影响

为提高枯草芽孢杆菌WSHDZ-01合成过氧化氢酶的水平,尝试了不同种类氮源的添加.结果表明,硝酸钠(NaNO3)为适宜氮源,虽然生物量仅1.25 g/L,但过氧化氢酶活力最高可达3 200 U/mL.在添加NaNO,的基础上,研究了添加其它氮源麦芽汁、酵母膏、玉米浆对提高wsHDz.0l生物量的影响,发现适宜浓度的麦芽汁不仅可以提高生物量,并且能够缩短发酵周期.经进一步优化,在3 L发酵罐中,WSHDZ-01生物量提高到6 g/L,过氧化氢酶活力达到11 000 U/mL,与优化前相比,发酵周期缩短了近60%,生产强度提高了3倍.     

产氢新种Ethanoligenens harbinese B49的氮素营养与代谢特征

从氮源角度研究了一株产氢新种Ethanoligenens harbineseB49发酵葡萄糖和糖蜜的产氢特性及其营养需求,以及酵母粉对产氢菌E.harbinese B49生长和产氢的特殊效应.试验结果表明,在以葡萄糖为底物条件下,以酵母粉替代蛋白胨作为唯一氮源可以大大提高E.harbineseB49的产氢能力,单位体积产氢量从1700mL L-1培养基提高到2400mL L-1培养基,产氢能力提高40.35%.该条件下去除维生素液,E.harbinese B49的产氢能力不受影响.从E.harbinese B49产氢动力学分析可以看出,对数生长期处于12~22h期间.从对数期开始迅速产氢,并且一直持续到稳定前期,产氢速率达到16.8mL/h,生物气中最大氢含量为41%.驯化后的E.harbinese B49利用糖蜜为底物产氢,糖蜜COD为13g L-1时,单位体积最大产氢能力为1576mL L-1.当培养基中额外加入0.5g L-1酵母粉时,可以大大促进E.harbinese B49发酵糖蜜产氢的能力,单位体积氢产量达到1960mL L-1,提高了24.4%,比产氢率达到150.8mL(H2)/g(COD).以牛肉膏、蛋白胨、尿素为氮源时,E.harbineseB49产氢量提高较小.图4表2参16     

Effects of nitrate concentration on biological hydrogen production by mixed cultures

The effects of nitrate on fermentative hydrogen production and soluble metabolites from mixed cultures were investigated by varying nitrate concentrations from 0 to 10 g N/L at 35°C with an initial pH of 7.0. The results showed that the substrate degradation rate, hydrogen production potential, hydrogen yield, and average hydrogen production rate initially increased with increasing nitrate concentrations from 0 to 0.1 g N/L, while they decreased with increasing nitrate concentrations from 0.1 to 10 g N/L. The maximum hydrogen production potential of 305.0 mL, maximum hydrogen yield of 313.1 mL/g glucose, and maximum average hydrogen production rate of 13.3 mL/h were obtained at a nitrate concentration of 0.1 g N/L. The soluble metabolites produced by the mixed cultures contained only ethanol and acetic acid (HAc) without propionic acid (HPr) and butyric acid (HBu). This study used the Modified Logistic model to describe the progress of cumulative hydrogen production in batch tests. A concise model was proposed to describe the effects of nitrate concentration on average hydrogen production rate.     

三峡库区笋溪河流域面源污染及其与土壤可蚀性k值的关系

研究三峡库区面源污染特征及其与水土流失的关系,可为库区氮磷污染和土壤侵蚀控制提供依据.选择三峡库区库尾笋溪河流域,在流域内分园地、林地和耕地3种土地利用类型共采集126个土壤样品,并在主干和支流采集52个水质样品.根据EPIC模型计算土壤可蚀性k值,分析流域内土壤可蚀性k值对面源污染的影响.结果表明,笋溪河流域面源污染主要是氮污染,总氮均值达1.37 mg/L,氮素的主要形态为硝态氮,占总氮的71.2%;总磷浓度为0.1 mg/L.流域内土壤可蚀性k值均值为0.040,随着土层加深土壤可蚀性k值呈上升趋势;林地土壤可蚀性k值显著低于园地和耕地.笋溪河流域总氮浓度与园地和耕地0-20 cm土壤可蚀性k值有关,硝态氮浓度与耕地0-40 cm土壤可蚀性k值有关.因此,笋溪河流域面源污染严重,主要来源是耕地和园地,应实行免耕、植物篱等措施,同时减少化肥施用,增加有机肥比例,以增加土壤抗侵蚀能力,进而控制流域水土流失和面源污染.(图6参37)     

Quantification of arginine requirements of juvenile marine shrimp,Penaeus monodon,using microencapsulated arginine

Using microencapsulated L-arginine, the quantitative requirement for amino acid has been determined for the first time for a shrimp species, which can not effectively utilize crystalline amino acids. In an 8 wk feeding trial (1990), juvenile Penaeus monodon were fed casein-based purified diets containing one of six levels (13.1, 17.7, 22.3, 26.9, 31.5 and 36.1 g/kg diet) of arginine. In addition to the protein-bound arginine already present in the casein of the test diets, pure arginine was supplemented by L-arginine microencapsulated in cellulose acetate phthalate, which is easily assimilated. The arginine level required for optimal growth was determined by brokenline model analysis of weight gain data to be 25.0 g/kg diet (=54.7 g/kg protein). Examination of the hemolymph 3 h after feeding revealed that the free arginine level in the hemolymph had not increased as a result of increasing levels of the dietary arginine. There was an abrupt increase of urea in the hemolymph when the arginine requirement of the shrimp had been met.     

Influence of glucose feeding on the ligninolytic enzyme production of the white-rot fungus Phanerochaete chrysosporium

The present work studied the influence of glucose feeding on the ligninolytic enzyme production of Phanerochaete chrysosporium in a nitrogen-limited (C/N ratio is 56/8.8 mmol/L) medium. Several sets of shaking flask experiments were conducted. The results showed that 2 g/L glucose feeding on the first day of the culture (24 h after the inoculation) simulated both fungal biomass growth and enzyme production. The manganese peroxidase (MnP) activity was 2.5 times greater than that produced in cultures without glucose feeding. Furthermore, the glucose feeding mode in fed-batch culture was also investigated. Compared to cultures with glucose feeding every 48 h, cultures with glucose feeding of 1.5 g/L (final concentration) every 24 h produced more enzymes. The peak and total yield of MnP activity were 2.7 and 3 times greater compared to the contrast culture, respectively, and the enzyme was kept stable for 4 days with an activity of over 200 U/L.     

Candida utilis生物合成谷胱甘肽的营养及环境条件

研究了摇瓶条件下Candida utilis WSH02—08生物合成谷胱甘肽(GSH)的营养条件，确定以葡萄糖作为碳源，硫酸铵和尿素作为混合氮源．在L6(4^5)正交试验的基础上，选用BP神经网络对营养条件进行优化和预测，得出较优的营养组合为：葡萄糖30gL^-1(NH4)2SO4gL^-1，尿素4gL^-1，KH2PO4 3gL^-1，MgSO4 0．25gL^-1．研究了GSH发酵的环境条件，得出较优的组合为：初始pH6．0，装液量30mL／250mL，接种量10％．利用优化后的营养及环境条件进行摇瓶发酵实验，结果表明，GSH产量和细胞干重均有较大幅度的提高．图7表2参13     

不同氮源对2种微藻生长及总脂含量的影响

采用一次培养的方式研究硝态氮（NaNO3）、氨态氮（NH4Cl）、有机氮（尿素）及硝铵混合氮（NH4NO3）（氮浓度17.6 mmol.L-1）对新分离的两株热带淡水微藻——网状空星藻Coelastrum reticulatum及栅藻Scenedesums sp.生长情况及总脂含量的影响。结果表明,2株微藻在不同氮源影响下生长状况不同,并且造成最终的干质量、总脂含量也有较为明显的差异。2种藻添加NaNO3和NH4Cl情况下分别获得了最高的生物量（（0.72±0.08）g.L-1和（0.80±0.03）g.L-1）和最高总脂含量（（38.35±1.32）%和（30.24±3.13）%）。最终二者在添加NaNO3的情况下单位体积总脂产量最高,分别为199.26 mg.L-1和190.76 mg.L-1,均可作为生物柴油的良好来源。     

Highly efficient and selective removal of phosphate from wastewater of sea cucumber aquaculture for microalgae culture using a new adsorption-membrane separation-coordinated strategy

● A new adsorption-membrane separation strategy is used for phosphate removal. ● PVC/Zr-BT shows a selective adsorption ability to low-concentration phosphate. ● Low concentration of P below 0.05 mg/L was achieved in actual wastewater treatment. ● Algal biomass production served as a demonstration of phosphorus recycling. Enhanced phosphorus treatment and recovery has been continuously pursued due to the stringent wastewater discharge regulations and a phosphate supply shortage. Here, a new adsorption-membrane separation strategy was developed for rational reutilization of phosphate from sea cucumber aquaculture wastewater using a Zr-modified-bentonite filled polyvinyl chloride membrane. The as-obtained polyvinyl chloride/Zr-modified-bentonite membrane was highly permeability (940 L/(m²·h)), 1–2 times higher than those reported in other studies, and its adsorption capacity was high (20.6 mg/g) when the phosphate concentration in water was low (5 mg/L). It remained stable under various conditions, such as different pH, initial phosphate concentrations, and the presence of different ions after 24 h of adsorption in a cross-flow filtration system. The total phosphorus and phosphate removal rate reached 91.5% and 95.9%, respectively, after the membrane was used to treat sea cucumber aquaculture wastewater for 24 h and no other water quality parameters had been changed. After the purification process, the utilization of the membrane as a new source of phosphorus in the phosphorus-free f/2 medium experiments indicated the high cultivability of economic microalgae Phaeodactylum tricornutum FACHB-863 and 1.2 times more chlorophyll a was present than in f/2 medium. The biomass and lipid content of the microalgae in the two different media were similar. The innovative polyvinyl chloride/Zr-modified-bentonite membrane used for phosphorus removal and recovery is an important instrument to establish the groundwork for both the treatment of low concentration phosphate from wastewater as well as the reuse of enriched phosphorus in required fields.     

Nitrogen and energy loss via nonfaecal and faecal excretion in the marine teleost Lithognathus lithognathus

Nitrogen excretion and assimilation efficiencies of individual Lithognathus lithognathus (Cuvier 1830), a marine teleost from high energy surf zones in Algoa Bay, South Africa, were determined under laboratory conditions in 1985. Nonfaecal-nitrogen excreted by starved and fed L. lithognathus consists mainly of ammonia with urea and amino acids as secondary excretory products. Ammonia excretion rates were temperature dependant with the excretion rate of starved fish significantly lower than those of fed fish, at all three experimental temperatures. The mass component b of the mass/ammonia excretion equation was temperature independent and ranged from 0.651 to 0.700 and 0.589 to 0.635 for starved and fed fish respectively. The mean percentage of food energy lost via dissolved nonfaecal excretory products (exogenous plus endogenous) was 6.11±6.07%. Assimilation efficiencies ranged from 70.75 to 99.29% for dry matter and from 95.72 to 99.58% on an energy basis. The combined nonfaecal and faecal energy loss was calculated at 11.87% of the ingested energy. The benthic feeding ichthyofauna recycle 255 g total nitrogen per metre strip per year which constitutes 2% of total phytoplankton nitrogen requirements of the surf zone.     

上海城市降水径流营养盐氮负荷及空间分布

在野外采样和室内化学分析的基础上，研究了上海市区，郊区各不同功能区内降水径流中营养元素氮的含量水平及其空间分布差异，研究结果表明，城市降水径流中营养元素氮具有较高的含量水平，其中TIN，NH4－N，NO3－N的均值含量分别为7．45，3．14，2．98和1．33mg/L，与同期河流水体中氮含量相比，降水径流中营养元素氮对河流水中氮含量水平具有重要的贡献，反映城市降水径流是一种不容忽略的非点污染源，从城市降水径流中营养元素氮负荷的空间分布来看，在工业区，交通要道，氮含量普遍偏高，而在居民居住区和商业区，氮含量普遍较低，此外，市区和郊区相比，在市区内氮含量普遍比郊区高。