Improvement of xylanase production by Aspergillus niger XY-1 using response surface methodology for optimizing the medium composition

Objective： To study the optimal medium composition for xylanase production by Aspergillus niger XY-1 in solid-state fermentation （SSF）. Methods： Statistical methodology including the Plackett-Burman design （PBD） and the central composite design （CCD） was employed to investigate the individual crucial component of the medium that significantly affected the enzyme yield. Results： Firstly, NaNO3, yeast extract, urea, Na2CO3, MgSO4, peptone and （NH4）2SO4 were screened as the significant factors positively affecting the xylanase production by PBD. Secondly, by valuating the nitrogen sources effect, urea was proved to be the most effective and economic nitrogen source for xylanase production and used for further optimization. Finally, the CCD and response surface methodology （RSM） were applied to determine the optimal concentration of each significant variable, which included urea, Na2CO3 and MgSO4. Subsequently a second-order polynomial was determined by multiple regression analysis. The optimum values of the critical components for maximum xylanase production were obtained as follows： x1 （urea）=0.163 （41.63 g/L）, x2 （Na2CO3）=-1.68 （2.64 g/L）, X3 （MgSO4）= 1.338 （10.68 g/L） and the predicted xylanase value was 14374.6 U/g dry substrate. Using the optimized condition, xylanase production by Aspergillus niger XY-1 after 48 h fermentation reached 14637 U/g dry substrate with wheat bran in the shake flask. Conclusion： By using PBD and CCD, we obtained the optimal composition for xylanase production by Aspergillus niger XY-1 in SSF, and the results of no additional expensive medium and shortened fermentation time for higher xylanase production show the potential for industrial utilization.     

Removal of Ammonia from Wastewater Effluent by Chlorella Vulgaris

The capability of Chlorella vulgaris to remove nitrogen in the form of ammonia and/or ammonium ions from wastewater effluent in a local wastewater treatment plant (i.e., the Mill Creek Plant in Cincinnati, Ohio, U.S.A.) was studied. The wastewater effluent leaving the plant was found to include high concentra-tions of nitrogen (7.7±0.19 mg/L) (ammonia (NH3) and/or ammonium ion (NH4+)) and total inorganic carbon (58.6±0.28 mg/L) at pH 7, and to be suitable for growing Chlorella vulgaris. When Chlorella vulgaris was cul-tivated in a batch mode under a closed system, half of the nitrogen concentration was dramatically removed in 48 h after a 24-h lag-phase period. Total inorganic carbon concentration also concomitantly decreased during the rapid growth-phase. The total biomass weight gained during the entire cultivation period balanced out well with the total amount of inorganic carbon and nitrogen removed from the culture medium. These results indicate that wastewater can be synergistically used to polish residual nutrients in wastewater as well as to cultivate microalgae for biofuel production.     

多粘类芽孢杆菌同步糖化发酵玉米粉生产(R,R)-2,3-丁二醇

【目的】对多粘类芽孢杆菌Paenibacillus polymyxa的玉米粉同步糖化发酵工艺进行优化,以获得低成本、高效的(R,R)-2,3-丁二醇生产技术。【方法】研究玉米粉浓度、氮源种类和氮源浓度对菌体生长、耗糖能力以及(R,R)-2,3-丁二醇产量、产率、得率和转化率的影响,并在此基础上进一步考察培养基中其它成分对(R,R)-2,3-丁二醇发酵的影响。【结果】优化后的培养基组分为玉米干粉140g/L,酵母粉30g/L,Na_2HPO_4 3g/L,KH_2PO_43g/L,(NH_4)2SO_42g/L,MgSO_40.8g/L,微量元素溶液2mL/L。使用优化后的培养基进行同步糖化发酵,发酵50h后(R,R)-2,3-丁二醇产量达到56.28g/L(光学纯度为98.3%),对葡萄糖的得率为0.44g/g,产率为1.13g/(L·h)。【结论】(R,R)-2,3-丁二醇生产技术低价高效,可为其工业化生产提供参考。     

毕赤酵母发酵生产重组人血清白蛋白高密度培养条件的研究

研究了毕赤酵母基因工程菌高密度培养条件。在全合成培养基的基础上考察了诱导过程中添加(NH₄)₂SO₄、微量元素PTM₁、油酸、甲醇加量及pH等因素对重组人血清白蛋白表达的影响，发现PTM₁能显著提高白蛋白的表达，6mL/L时蛋白表达量最大。诱导期硫酸铵浓度维持在 3g/L左右蛋白表达量最大，高于3g/L抑制蛋白表达。酸性环境不利于蛋白表达，诱导过程维持pH6.0～6.5最佳。甲醇最佳诱导浓度为10mL/L。添加0.05%的油酸可有效提高蛋白表达。可以此作为发酵罐上发酵培养流加成分的参考。     

维生素对红法夫酵母产虾青素的影响

采用单因素实验方法研究了不同维生素对红法夫酵母(Phaffia rhodozyma)生长及虾青素积累的影响.结果表明:生物素、泛酸钙、肌醇、VB1可显著提高红法夫酵母的生物量、类胡萝卜素和虾青素的产量,其较适质量浓度分别为0.05,25.00,0.50,0.05 mg/L.VB12和核黄素对红法夫酵母的细胞生长有明显的促进作用,同时可略微提高虾青素产量,其较适质量浓度分别为0.50和0.005 mg/L;VB6、烟酸和叶酸虽然可促进细胞生长,但却明显抑制了虾青素的积累;而硫辛酸既抑制细胞生长也抑制其虾青素的积累.根据单因素实验结果,在培养基中按以上较适浓度添加生物素、泛酸钙、肌醇、VB1、VB12和核黄素,菌体生物量和虾青素质量浓度分别达到5.000 g/L和16.77 mg/L,较对照组分别提高了33.2%和108%.     

生物素对红发夫酵母生长和虾青素积累的影响

在摇瓶中研究生物素对红发夫酵母生长和虾青素积累的影响.结果表明生物素的添加能促进细胞生长,但对细胞内虾青素含量的提高不利.添加10μg/L生物素时,最大比生长速率达到0.110 h-1;添加6μg/L生物素时,达到最大细胞干重9.95g/L,比没有添加生物素时的7.53g/L提高32%.添加2μg/L生物素时,达到最高虾青素产量和产率,分别为3 825 μg/L和53μg/(L@h),比没有添加生物素时的2 944 μg/L和41μg/(L@h)提高30%.没有添加生物素时,达到最高细胞虾青素含量391μg/g.利用添加2μg/L生物素的方法可以提高虾青素产量,这种方法为提高虾青素产量提供了一种简单廉价的方法.     

2-脱氧葡萄糖对酵母虾青素积累及酶法提取的影响

2-脱氧葡萄糖是一种酵母细胞壁合成抑制剂,红发夫酵母培养中添加0．0005％—0．002％（w／v）的2-脱氧葡萄糖可使生物量基本保持不变,但能促进虾青素积累,在增加虾青素产量的同时缩短酶法提取的时间。当添加浓度为0．002％时,虾青素含量可提高到对照组的113．7％,若采用胞壁溶解酶对红发夫酵母破壁提取虾青素,作用12h提取率可达94．7％。同时,2-脱氧葡萄糖应在红发夫酵母培养的前36h添加。     

Optimization of butanol production from corn straw hydrolysate by <Emphasis Type="Italic">Clostridium acetobutylicum</Emphasis> using response surface method

Butanol is a new kind of very potential biofuels.Enzymatic hydrolysis of corn stalk was utilized in this study to produce butanol by Clostridium acetobutylicum CICC 8008.Plackett-Burman (P-B) design and Central Composite Design (CCD) were adopted to screen crucial factors during fermentation as well as the optimization of experimental conditions.The result demonstrated that among the seven factors,namely,Yeast extract,(NH 4) 2 SO 4,KH 2 PO 4,MgSO 4,FeSO 4,CuSO 4 and CaCO 3,only CaCO 3 was selected as the most critical factor.The optimization experiment results for CaCO 3 usage,temperature and reaction time by CCD were determined to be 5.04 g/L,35°C and 70 h,respectively.A corresponding mathematical model was established to predict the fermentation experiment and maximum butanol yield of 6.57 g/L was acquired.The result of verification experiment under the optimum conditions showed that 6.20 g/L was the maximum butanol yield.This demonstrated that statistical method was a powerful tool for the optimization of butanol production from enzymatic hydrolysis of corn stalk.     

正交设计法筛选槐耳菌丝体液体培养基的研究

以槐耳菌丝体生物量为主要指标,采用不同碳源和氮源进行单因素初选试验,然后以不同比例的混合碳氮源进行正交试验．结果表明,槐耳菌丝体液体最佳培养基是玉米渣200g／L,蔗糖20g／L,酵母膏3g／L,麦麸3．5g／L,KH2PO42．0g／L,MgSO4·7H2O1．0g／L和VB1 6mg／L,温度28℃,pH自然．其菌丝体生物量得率为21．6g／L．     

乙醇/硫酸铵双水相体系分离纯化款冬花总黄酮

以低温烘干的款冬花为原料,利用乙醇/硫酸铵双水相体系分离纯化款冬花总黄酮。首先对无水乙醇、(NH_4)_2SO_4、黄酮粗提取液、NaCl质量分数和pH值进行单因素研究,然后利用Box-Benhnken Design实验对(NH_4)_2SO_4质量分数、无水乙醇质量分数、黄酮粗提取液质量分数、pH值进行了优化;结果表明:(NH_4)_2SO_4质量分数为15.5%、无水乙醇质量分数为32.2%、黄酮粗提取液质量分数为20.4%、pH=4.1时款冬花总黄酮萃取率可达99.21%,与预测的99.44%非常接近。     

裂殖壶菌Schizochytrium sp.FJU-512细胞油脂的研究

裂殖壶菌Schizochytrium sp.FJU-512是高产二十二碳六烯酸(DHA)的一株海洋真菌,其细胞内油脂占细胞干质量的18.3%.细胞油脂的总脂肪酸组成为45.7%DHA,12.5%DPA和41.8%棕榈酸.总油脂由26.4%极性脂和73.6%中性脂组成.薄层色谱、高效液相色谱及质谱分析表明该菌株还能合成强抗氧化剂虾青素.采用均匀设计获得高产DHA的最优培养基(g/L):葡萄糖109.7,蛋白胨19.2,酵母膏20.0,海水晶10.4,DHA产量高达2.10 g/L.     

Characteristics of electric double layer in different aqueous electrolyte solutions for supercapacitors

Aiming to find suitable electrolytes for electric double layer capacitor (EDLC),the relationship between the formation velocity of electric double layer and the specific capacitance in different electrolyte (H 2 SO 4,KOH,KCl,NH 4 Cl,and LiOH) solutions was investigated by cyclic voltammetry and AC impedance with varying frequency and potential,which was also analyzed based on Stern model and Gouy-Chapman diffusion layer model.It shows that the capacitance is positively correlated with the formation velocity of the electric double layers,which is affected by the type of ions applied,the concentration of the electrolyte solution,and the potential and frequencies as well.The electrolyte solutions of 6 mol/L KOH and 4 mol/L H 2 SO 4 provide the largest capacitance,reaching 214 and 186 μF/cm 2,respectively.Both 6 mol/L KOH and 4 mol/L H 2 SO 4 solutions are suitable electrolytes for EDLC applications using Pt as electrodes.     

Ammonium improves iron nutrition by decreasing leaf apoplastic pH of sunflower plants (<Emphasis Type="Italic">Helianthus annuus</Emphasis> L. cv. Frankasol)

The effect of nitrogen form on pH and concentration of soluble iron (Fe) in leaf apoplast was investigated in hydrophonically grown sunflower plants (Helianthus annuus L. cv. Frankasol), and the mechanism underlying the improved Fe nutrition by ammonium (NH₄) supply was also elucidated. Ammonium supply ameliorated Fe nutrition of plants grown without Fe through decreasing apoplastic pH and increasing soluble Fe concentration in apoplastic fluid of young leaves. The soluble Fe concentration in apoplastic fluid and cell sap of young leaves, and xylem exudates of NH₄ fed-plants was higher than that of nitrate (NO₃) fed-plants, and no typical Fe-deficiency chlorosis in young leaves was observed in NH₄, fed plant without Fe supply. The apoplastic pH was 6.15 and 5.94 in young leaves of Fe-deficient plants fed respectively with NO₃ and NH₄, while in Fe-sufficient plants, the apoplastic pH was 6.43 with NO₃, and 5.50 with NH₄ supply. In primary leaves, the apoplastic pH was around 6.25 irrespective of nitrogen form and Fe supply. The pH of xylem exudate was 5.72 in Fe-deficient plants fed with NO₃ and 5.49 with NH₄. Iron nutrition increased the pH of xylem exudate by 0.27 and 0.16 unit under NO₃ and NH₄ supply respectively.     

黑芝菌丝体液体培养基的筛选

以黑芝菌丝体生物量为主要指标,首先对不同碳源和氮源进行单因素初选试验,然后进行不同比例混合碳氮源正交试验.结果表明：黑芝菌丝体液体最佳培养基是蔗糖20 g/L、麦芽糖15 g/L、麦麸20 g/L、酵母粉1.5 g/L、KH2PO42 g/L、MgSO4·7H2O 1 g/L和VB16 mg/L,其菌丝体生物量产率为35 g/L.     

三角褐指藻的培养条件研究

对三角褐指藻(Phaeodactylum tricornutum)进行了培 养条件研究,结果表明,在20 ℃下,采用去除氮源的No.4基本培养液,起始pH值为8.0,(N H2)2CO为氮源,氮浓度为0.01 mol/ L时,对藻细胞的光合自养生长极为有利,培养10 d,其生物量达441 mg/L.过高的碳酸氢钠 浓度(>0.04 mol/L)易引起培养液pH值的过大改变而导致培养液中出现沉淀,在培养液中加 入葡萄糖有利于藻细胞的生长,光合自养生长生物量浓度较低,混合营养明显促进藻细胞的 生长,且培养液中同时含质量分数0.2%葡萄糖和0.04 mol/L碳酸氢钠后藻生物量达1 630 mg /L,因此光合异养生长明显促进藻细胞的生长.     

Study on screening and cultivation conditions of xylanase-producing alkalophilic bacterial

Xylanisthemajorcomponentofplantcellwallsandthemostabundantpentosaninnature.Xylancontainsβ 1,4 linkedd xylosebackboneswitharabinose,4 O methyl d glucuronicacid,andaceticacidsubstituents[1 ] .Thexylan degradingen zymesincludeendoxylanase(1,4 β d xylanxylanohydrolase;EC3.2 .1.8)and β xylosidase (1,4 β d xylanxylanohydrolase;EC 3.2 .1.37) .xylanasedegradesthexylanbackboneintoxylo bioseandxylooligosaccharides,whileβ xylosidasereleasesxylosylresiduesfromthenonreducingendsofxylooligosaccha…     

多因子正交试验对发根农杆菌生物波培养基的筛选

用Ｌ１６（４＾５）正交设计考察了牛肉浸膏、酵母浸出汁、蛋白胨、蔗糖及琼脂５种ＶＥＢ细菌培养基成分对发根农杆菌Ａ４的生物波形成的影响,主差分析结果显示,各因子对菌落直径的影响都极显著,它们的作用大小依次为：琼脂,蛋白胨,蔗糖,牛肉浸膏,酵母梁出汁,各因子对生物波成环数的影响也都极显著,它们的作用大小依次为：蔗糖,牛肉浸膏,蛋白胨,酵母浸出讦,琼脂,筛选出的最佳波动波动培养基为２．０ｇ／Ｌ牛肉浸膏＋０     

重组脂肪酶自诱导发酵培养基优化

【目的】优化自动诱导发酵培养基,提高脂肪酶产量。【方法】通过对构建的一株转座子整合型脂肪酶重组菌株,采用自动诱导发酵表达方法,并结合CCD响应面实验设计方法,对自动诱导培养基中的碳源进行优化,获得了1个二次模型用于描述发酵培养基中碳源对产脂肪酶的影响。【结果】优化后的最适自动诱导培养基(W/V)组成为glycerol 2.596,glucose 0.035,lactose 1.289,tryptone 1.0,yeast extract 0.5,Na_2HPO_4·12H_2O 1.79,KH_2PO_4 0.68,NH_4Cl 0.267 5,Na_2SO_4 0.071,MgSO_4·7H_2O 0.05。【结论】发酵试验表明,最优碳源培养基的比活力为R_1=6.35U/mg,比初始发酵培养基提高近3倍。     

一株致香真菌产香碳、氮源的优化

以菌落直径及致香程度为指标,分别采用不同碳源、氮源对一株致香真菌进行平板培养,筛选出致香真菌产香及生长的最佳碳源、氮源。结果表明:该致香真菌最适N源、C源依次为蛋白胨和葡萄糖,确定致香菌株最优的培养基配方为蛋白胨2 g/L,K2HPO41 g/L,KCl 0.5 g/L,Mg SO4·7H2O 0.5 g/L,Fe SO4·7H2O 0.01 g/L,葡萄糖3 g/L,麸皮3 g/L。     

Reaction condition optimization and kinetic investigation of roasting zinc oxide ore using (NH<Subscript>4</Subscript>)<Subscript>2</Subscript>SO<Subscript>4</Subscript>

An orthogonal test was used to optimize the reaction conditions of roasting zinc oxide ore using (NH₄)₂SO₄. The optimized reaction conditions are defined as an (NH₄)₂SO₄/zinc molar ratio of 1.4:1, a roasting temperature of 440°C, and a thermostatic time of 60 min. The molar ratio of (NH₄)₂SO₄/zinc is the most predominant factor and the roasting temperature is the second significant factor that governs the zinc extraction. Thermogravimetric–differential thermal analysis was used for (NH₄)₂SO₄ and zinc mixed in a molar ratio of 1.4:1 at the heating rates of 5, 10, 15, and 20 K·min-1. Two strong endothermic peaks indicate that the complex chemical reactions occur at approximately 290°C and 400°C. XRD analysis was employed to examine the transformations of mineral phases during roasting process. Kinetic parameters, including reaction apparent activation energy, reaction order, and frequency factor, were calculated by the Doyle–Ozawa and Kissinger methods. Corresponding to the two endothermic peaks, the kinetic equations were obtained.