玉米秸秆稀酸-蒸汽爆破预处理和水解糖化的试验研究

考察了玉米秸秆经1%(w/w)稀硫酸和水分别浸泡后在不同汽爆压力(分别确定研究压力为1.5 MPa,1.8 MPa和2.0 MPa)和保压时间(分别为4 min,6 min和8 min)下进行蒸汽爆破预处理的处理效果。分析了预处理后固体和液体部分的主要成分和含量。通过考察预处理后固体部分经过纤维素酶作用后所得到的葡萄糖得率,确定了最佳的稀酸-蒸汽爆破预处理工艺。在1%稀硫酸预浸12 h后,采用1.8 MPa汽爆条件保压8 min,经过预处理玉米秸秆的最大葡萄糖得率为26.9 g/100g原料;在该条件下,预处理后过滤液中总糖得率最高为34.5 g/100 g原料。     

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.     

固定化混合菌种发酵玉米秸秆水解液的研究

通过固定化混合菌种发酵法,研究了嗜单宁管囊酵母（Pachysolen tannophilus）和酿酒酵母（Saccharomyces cerevisiae）共固定于海藻酸钙中发酵玉米秸秆水解液工艺。研究结果表明,所得到的固定化混合菌种能同时利用玉米秸秆水解液中的木糖和己糖共同发酵,其效果明显优于游离混和菌种发酵,也明显优于S.cerevisiae和P.stipitis两种细胞共固定发酵。在发酵周期24h,初始pH值6.0,初始糖浓度118.42g/L的条件下,酒精转化率可达到0.4g/g糖。     

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。     

玉米秸秆水解的酶法与稀酸法比较

探讨玉米秸秆在纤维素酶及稀酸作用下的水解方法,并从水解影响因素（水解时间、温度、底物浓度等）及水解机理上,比较了两种纤维素酶与稀硫酸对玉米秸秆水解特性.结果表明：由于酶和酸的水解机理不同,对玉米秸秆的水解影响也不一样,酶水解速度慢,水解得率高,条件温和;稀酸水解速度快,水解得率低,对设备要求高.如果酸和酶结合,则玉米秸秆水解得率有很大的提高.     

玉米秸秆瞬间蒸汽爆破预处理的回归优化和结构分析

采用响应面法回归分析优化瞬间蒸汽爆破预处理玉米秸秆过程,研究了汽爆压强、维压时间以及填料量三因素对酶解糖产率的影响,基于Box-Behnken设计,分析并获得了一个二阶线性方程模型,能够较好地拟合实验值。获取的最优条件为汽爆压强3.5MPa,维压时间50s,填料量60g,此时糖产率达到54.37%,相比于未处理物料,其糖化率提高了1.88倍。采用扫描电镜、X射线衍射分析以及傅里叶红外光谱对处理前后的物料进行结构和组分分析,与未处理的物料相比,处理后的物料结晶度明显降低,颗粒度减小,可及度显著提高。     

Study on synthesis of polylactide from kitchen garbage

Polylactide, a kind of synthetical aliphatic polyesters, has excellent biologic compatibility and biodegradable properties. It can be degraded in the human body by sim-ple hydrolysis of the ester backbone to non-harmful and non-toxic compounds, and finally be biodegraded into CO2 and H2O in the environment. Therefore, for many years, the polylactide is widely used as drug delivery sys-tems, resorbable suture materials and implantable medical devices. Besides medical applications, it will be …     

Cellulose-hydrogen production from corn stalk biomass by anaerobic fermentation

Cellulose-hydrogen production from corn stalk by lesser panda manure was carried out in batch tests and a 5 L scale-up continuously stirred anaerobic bioreactor(CSABR),respectively.The bio-pretreatment of corn stalk was found most effective at 25℃ using microbe additive of 7.5 g/kg,in which the yields of soluble saccharides(SS) and lactic acid were 212 mg/g-TS and 21 mg/g-TS,respectively.The maximum cumulative H2 yield(176 ml/g-TS) and H2 production rate(14.5 ml/g-TS h-1) were obtained at pH 5.5,36℃ by trea...     

德氏乳杆菌保加利亚亚种培养基及培养条件的优化

为了降低德氏乳杆菌保加利亚亚种的工业生产成本和提高发酵液的菌浓度,通过单因素试验和旋转中心组合设计（central composite design,CCD）相结合的方法优化培养基的组分和培养条件。优化后的培养基成分为:葡萄糖30 g/L、豆粕28 g/L、玉米粉14 g/L、乳清粉28 g/L、K2HPO4 3.0 g/L、柠檬酸三铵2.5 g/L、乙酸钠5 g/L、吐温-80 1.25 mL/L、MgSO4·7H2O 0.2 g/L、MnSO4·4H2O 0.0625 g/L;培养条件为:初始pH值7.1,温度37 ℃,接种量4%（体积分数）,静置培养。经优化后活菌数达到 6.07×109 CFU/mL,明显高于原MRS培养基（5.8×108 CFU/mL）,且其成本较原MRS培养基的成本降低了4000元/t。     

利用曲霉sp.HX-1发酵稻草秸秆制备纤维素酶

以稻草秸秆原料,利用曲霉sp.HX-1固态发酵生产纤维素酶,研究了不同发酵时间、不同氮源和氮源浓度对曲霉sp.HX-1纤维素酶系的影响,并最终用硫酸铵分级沉淀和低温冷冻干燥的方法得到混合纤维素酶干品.结果表明,发酵6 d后稻草秸秆产生的纤维素酶活力单位最大,分别为CMC酶307 U/mL,C1酶841 U/mL、β-葡萄糖苷酶205 U/mL.以不同氮源优化发酵条件时发现,NH4NO3质量浓度为1 g/L时CMC酶活力达到1 652 U/mL,且失重率也到达最大值17.42%;NH4Cl质量浓度为0.5 g/L时,C1酶活力达到1 807 U/mL;尿素（UREA）质量浓度为2.0 g/L时,β-葡萄糖苷酶活力为2 033 U/mL,这表明氮源对曲霉sp.HX-1纤维素酶系的影响很大.最后在NH4NO3质量浓度为1.0 g/L的条件下,将120 g稻草秸秆发酵6 d,从发酵液中提取得到8.851 7 g混合纤维素酶的干品.此实验为以后探讨碳源或者其他因素的影响提供方法借鉴,也可以为获得纤维素酶混合酶干品的获得提供参考方法.     

高耐受性拜氏梭菌的离子束诱变选育

【目的】为了优化生物丁醇生产工艺,研究拜氏梭菌(Clostridium beijerinckii)对有毒物质的耐受性。【方法】利用常温等离子诱变技术选育获得1株突变株C.beijerinckii GZ-9,并通过摇瓶发酵对比原始菌株C.beijerinckii NCIMB 8052和突变株GZ-9对有毒物质的耐受性。【结果】NCIMB 8052在酚浓度为1.5g/L的发酵液中已不能生长,而GZ-9在酚浓度2.4g/L的发酵培养基中生长良好;当可溶性总酚浓度为1.5g/L时,实验总溶剂产量和丁醇产量分别达9.2g/L和6.5g/L。【结论】突变株GZ-9对未脱毒甘蔗渣酸解糖液中毒素物质的耐受性远高于原始菌株NCIMB 8052。     

Effects of soil stoichiometry of the CaCO<Subscript>3</Subscript>/available phosphorus ratio on plant density in <Emphasis Type="Italic">Artemisia ordosica</Emphasis> communities

In soil, CaCO3 can reduce the availability of phosphorus in arid and semi-arid regions, thereby influencing plant growth. However, the effects of soil stoichiometry of the CaCO3/available phosphorus ratio (CaCO3/AP ratio) on plant density are unknown. In this study, we examined 4 Artemisia ordosica communities located in arid and semi-arid regions, and performed a pot experiment with an orthogonal design to examine the relationships between the soil CaCO3/AP ratio and plant density. The results showed that the soil stoichiometry of the CaCO3/AP ratio had a greater effect on growth of A. ordosica than either CaCO3 or AP as single factor. The influence of the soil CaCO3/AP ratio on density of A. ordosica was related to the climatic zone and the types and/or amplitudes of increases in the soil CaCO3/AP ratio. When the soil CaCO3/AP ratio was within the same amplitude and was less than 2.5, the A. ordosica density increased in the semi-arid region, whereas the density decreased with increasing soil CaCO3/AP ratios in the arid region. In the semi-arid region, when the soil CaCO3/AP ratio was within the same amplitude and the increase in the soil CaCO3 content was greater than the increase in soil AP, the A. ordosica density increased with increasing soil CaCO3/AP ratios. This means that the relationship between the soil CaCO3/AP ratio and the A. ordosica density remained the same. In the arid region, an increase in the soil CaCO3/AP ratio as a result of increased soil CaCO3 content and decreased soil AP resulted in increased density of A. ordosica. Therefore, the relationship between soil CaCO3/AP ratios and density of A. ordosica was altered by these changes in soil chemistry. Thus, the soil stoichiometry of the CaCO3/AP ratio is an important factor affecting the density of A. ordosica in arid and semi-arid regions.     

预处理玉米秸秆中抑制物对酶解与发酵影响研究

玉米秸秆在预处理过程中会生成酵母抑制物,不利于发酵产乙醇。本文考察了七种模型抑制物对酵母菌发酵乙醇的影响,并对预处理玉米秸秆（PCS）进行了脱毒试验,研究了酶解与同步糖化发酵过程。得到,对酿酒酵母FE-B抑制作用由强到弱为苯系化合物＞呋喃衍生物＞弱酸类;水洗至中性后挤压PCS的酶解和发酵效果均好于未处理和挤压处理PCS,在固含量为30%的酶解与发酵体系中,72 h葡萄糖浓度为12.23%,发酵24 h乙醇浓度达到5.82 w%。     

一株高温蛋白酶高产菌株产酶条件的优化

对一株高温蛋白酶高产菌株枯草芽孢杆菌BY25的发酵培养基组成与产酶条件进行了优化。正交试验结果显示培养基中各因子对产酶影响从高到低为:豆饼粉、葡萄糖、硫酸镁、麸皮、磷酸氢二钠、氯化钙。在此基础上进行培养基组成优化,将豆饼粉、麸皮混合氮源改为以豆饼粉为单一氮源进行蛋白酶发酵。单因素试验发现,在单一氮源培养条件下,培养基中各因子对产酶影响从高到低依次为:豆饼粉、葡萄糖、氯化钙、磷酸氢二钠。除微量氯化钙外,金属盐,尤其是金属硫酸盐的添加对产酶有显著抑制作用。此外,培养初始pH和培养时间对产酶有显著影响,接种量也有一定影响。通过绘制120 h产酶曲线发现,BY25产酶曲线为双峰,产酶曲线顶峰出现在发酵后72 h,优化后BY25发酵培养基各组分添加量为豆饼粉60 g/L、葡萄糖60 g/L、氯化钙 0.5 g/L、磷酸氢二钠 4 g/L,接种量为4.5%～5%,初始培养pH为8.0。优化产酶培养基和产酶条件后,发酵液酶活力可达到101.1 μmol/(min·mL)。     

响应面法优化萎缩芽孢杆菌BsR05发酵培养基

为了提高萎缩芽孢杆菌Bacillus atrophaeus BsR05发酵液的芽孢产量,采用响应面法对BsR05发酵培养基的最佳工艺条件进行了优化。通过Plackett-Burman实验,筛选出玉米粉、(NH_4)_2SO_4和MgSO_4·7H_2O为影响产孢的主要因子。采用最陡爬坡路径法确定3个因素的响应中心点及最适浓度范围,最后,通过Box-Behnken设计建立主要培养基成分与芽孢产量之间的回归关系,并确定发酵培养基最佳配方为葡萄糖5 g/L、玉米粉15.9 g/L、豆粕40.0 g/L、K_2HPO_4 3.0 g/L、KH_2PO_4 1.0 g/L、(NH_4)_2SO_4 2.1 g/L、MgSO_4·7H_2O 0.40 g/L、MnSO_4 0.02 g/L。经重复实验验证,平均芽孢含量与预测芽孢含量基本一致,发酵液中BsR05的芽孢产量从优化前的4.73×10~9 CFU/m L提高到6.02×10~9 CFU/m L。     

Studies on optimization of nitrogen sources for astaxanthin production by Phaffia rhodozyma

Fermentation of Phaffia rhodozyma is a major method for producing astaxanthin, an important pigment with industrial and pharmaceutical application. To improve astaxanthin productivity, single factor and mixture design experiments were used to investigate the effects of nitrogen source on Phaffia rhodozyma cultivation and astaxanthin production. Results of single factor experiments showed nitrogen source could significantly affect P. rhodozyma cultivation with respect to carbon source utilization, yeast growth and astaxanthin accumulation. Further studies of mixture design experiments using (NH₄)₂SO₄, KNO₃ and beef extract as nitrogen sources indicated that the proportion of three nitrogen sources was very important to astaxanthin production. Validation experiments showed that the optimal nitrogen source was composed of 0.28 g/L (NH₄)₂SO₄, 0.49 g/L KNO₃ and 1.19 g/L beef extract. The kinetic characteristics of batch cultivation were investigated in a 5-L pH-stat fermentor. The maximum amount of biomass and highest astaxanthin yield in terms of volume and in terms of biomass were 7.71 mg/L and 1.00 mg/g, respectively.     

CaCO<Subscript>3</Subscript> film synthesis from ladle furnace slag: morphological change,new material properties,and Ca extraction efficiency

A rapidly air-cooled ladle furnace slag (RA-LFS), which is a type of steelmaking slag discharged from a steel mill, was used to synthesize CaCO₃ film. The CaCO₃ film with 35 cm2 of surface area was synthesized under atmospheric conditions, and the surface morphology of the CaCO₃ films was changed by using additives (CaCl₂ and ethylene glycol). Especially, the addition of CaCl₂ changed the surface morphology of CaCO₃ film with pore and induced new material properties, such as water adsorption. The (012) face of CaCO₃ film (calcite) was rapidly decreased by the addition of CaCl₂. The major components of RA-LFS were calcium (type of CaO, 53.9wt%) and aluminum (type of Al₂O₃, 37.9wt%), and the major crystal phases of RA-LFS were C₃S, C₁₂A₇, and C₃A. The calcium extraction efficiency of RA-LFS was significantly increased after the CaCO₃ film synthesis. The material properties (hardness and elastic modulus) and the thermal characteristics of the CaCO₃ films were analyzed by nano-indentation and thermogravimetry-differential thermal analysis. The synthesized CaCO₃ films from RA-LFS and Ca(OH)₂ (reagent) showed similarities in terms of their material properties and the decomposition temperature.     

餐厨垃圾发酵产油脂的复合酶制剂水解试验

为使餐厨垃圾能有效地通过微生物发酵产油脂,采用复合酶对餐厨垃圾进行水解预处理研究。研究结果表明,复合酶中淀粉酶、糖化酶、蛋白酶和纤维素酶投加量分别为92.5,1 250,3 000,100活力/g原料时,效果最优;其水解产物中氨基酸态氮浓度为137.7mg/L,还原糖的质量百分比含量为11.11%。在复合酶各组分投加量一定时,水解效果受pH值、温度和水解时间影响。试验结果表明,pH值为6、温度为55℃、水解时间为30min时,水解效率最高;在最佳水解条件下,每升水解液经从土壤筛选分离的菌种B发酵7d后,生物产量36.9g,微生物油脂产量5.172g。     

产丁醇重组大肠杆菌的构建及初步发酵

将丙酮丁醇梭菌ATCC 824丁醇合成途径中分别编码硫解酶(THL)、3-羟基丁酰-CoA脱氢酶(HBD)、巴豆酸酶(CRT)、丁酰-CoA脱氢酶(BCD)和醛/醇脱氢酶(BDH)的关键基因thil、hbd、crt、bcd-etfB-etfA、adhE在大肠杆菌中进行重组表达,构建出3株产丁醇的重组大肠杆菌JM109-99d、DL1346 99d和AFP111-99d。选取摇瓶发酵产丁醇最高(0.52 g/L)的重组大肠杆菌DL1346-99d进行上罐发酵,丁醇产量可达1.07g/L。     

香蕉杆组分测定及其产沼气潜力的研究

【目的】测定香蕉杆中的纤维素、半纤维素和木质素组分,并对其发酵产沼气的潜力进行研究,以期为香蕉杆的资源化利用提供可靠的基础资料。【方法】分别采用改进后的硫酸-重铬酸钾氧化法、2mol/L盐酸水解法和72%浓硫酸法测定香蕉杆中纤维素、半纤维素和木质素的含量;在35℃恒温条件下进行厌氧发酵产沼气,绘制不同原料产沼气过程变化曲线图。【结果】香蕉杆纤维素、半纤维素和木质素含量分别为17.47%、11%、6.86%,其厌氧发酵产沼气最高可达到(155±3)mL/g TS。【结论】香蕉杆纤维含量较高,可作为沼气发酵原料使用,为香蕉杆的资源化利用提供有益参考。