硅橡胶膜生物反应器封闭循环连续发酵制造乙醇放大实验及该发酵系统的基本性能

设计构建了发酵罐体积为5 L,单张膜有效面积为0.08 m2的平板硅橡胶膜生物反应器封闭循环连续发酵系统,实验研究了该系统在长期运行过程中的发酵反应动力学参数和膜传质动力学参数等基本性能.当发酵罐中乙醇浓度在30～60 g·L-1时,得质量浓度为17% ～28%的冷凝渗透液.在连续运行中,细胞浓度维持在10～24.8 g·L-1,料液罐中葡萄糖浓度大约为30～50 g·L-1,乙醇的体积产率为2.33～3.99 g·L-1·h-1,膜的渗透通量和分离因子分别为800～1050 g·m-2·h-1和5.1～8.6. 在连续269 h运行中,得到乙醇1999 g,基质转化率为87.2%,碳回收率为89.5%,产生的废液量大约为传统间歇发酵过程的22.2%左右.     

乙醇连续发酵-渗透汽化耦合系统发酵动力学研究

利用自制硅橡胶平板复合膜构造乙醇连续发酵-渗透汽化耦合系统,研究了系统长期运行中发酵反应动力学问题,并分析了连续发酵过程中细胞比生长率的变化。系统稳定运行了269h。由于渗透汽化的进行,发酵液中乙醇浓度控制在35—45g·L-1,酿酒酵母浓度维持在20—25g·L-1,在发酵与渗透汽化耦合阶段,乙醇产率达到3.4g·h-1L-1,是间歇发酵时的4.5倍。乙醇得率系数为0.46,乙醇的转化率达到91%。膜下游冷凝收集液的浓度可达28.2—16.5%(wt),总渗透通量达到1226—707g·m-2h-1,乙醇渗透通量为293—117g·m-2h-1,分离因子为8.5—4.9。     

自絮凝颗粒酵母发酵菊芋汁生产乙醇

分别采用分批和连续发酵方式,对自絮凝颗粒酵母Saccharomyces cerevisiae flo发酵菊芋汁生产乙醇的条件进行了优化. 与先酶解菊芋汁后再用自絮凝酵母发酵的分步糖化发酵相比,分批发酵过程中同时加入菊粉酶和自絮凝酵母的同步糖化发酵乙醇得率高,发酵时间短. 当菊芋汁总糖浓度分别为105和179 g/L时,同步糖化发酵的最高乙醇浓度达50和82.5 g/L,比分步糖化发酵高6.4%和13.8%. 在连续发酵过程中应用同步糖化发酵法,当稀释率为0.02 h-1时,乙醇浓度约为90 g/L时达到稳定状态,乙醇得率达到理论值的90%,生产强度达2.12 g/(L×h).     

乙醇发酵与渗透汽化在硅橡胶膜生物反应器中的耦合强化

用硅橡胶膜生物反应器(SMBR)实验研究了发酵-渗透汽化的耦合性能。发酵微生物采用酿酒活性干酵母,所用的碳源为工业级葡萄糖。间歇发酵过程由于产物抑制作用在乙醇浓度达到90g稬-1时就趋于停滞,而经耦合渗透汽化膜分离后,发酵罐内的乙醇浓度迅速降低并维持在40g稬-1,且发酵在此浓度下可以连续稳定地进行。 在SMBR运行达到稳态后,乙醇的体积产率为1.5gL-1h-1。SMBR中所用的聚二甲基硅氧烷(PDMS)复合膜由实验室自行制备,它能稳定分离含有酵母细胞的发酵液。当发酵液中乙醇浓度为92.7~49.5g稬-1时,PDMS复合膜的总通量为1490~1164g穖-2h-1,分离因子为6.9~7.8,与分离相同进料浓度的清洁模型溶液相比分别平均高出31%和14%。乙醇发酵和渗透汽化在硅橡胶膜生物反应器中能够相互耦合并得到强化。     

LSM蛋白增强木糖发酵重组酵母抗逆性能的研究

木质纤维素原料预处理过程中产生的弱酸、呋喃醛类和酚类化合物等对酿酒酵母的乙醇发酵有抑制作用,提高基因重组酵母对抑制物的耐受性,是利用植物秸秆水解液生产燃料乙醇的关键技术之一。研究从前期构建的戊糖、己糖共发酵重组酿酒酵母Saccharomyces cerevisiae ZU-E8基因组DNA中克隆出RNA结合蛋白LSM6,将其连入含有PADH启动子的质粒构成表达载体pR-LSM,进而转入ZU-E8宿主细胞中。通过高浓度醋酸根平板筛选,得到高抗逆性木糖发酵重组酵母ZU-910。在醋酸浓度为2 g·L-1的木糖培养基中发酵96 h后,ZU-910的木糖利用率和乙醇浓度为90.2%和26.9 g·L-1,分别是出发菌株ZU-E8的8.5和10倍,并且ZU-910对糠醛和硫酸根的耐受能力也较ZU-E8大大增强。在玉米秸秆酶解液发酵中,ZU-910的木糖利用率和乙醇产量在ZU-E8基础上增加了10.5%和7.7%.证明LSM6蛋白确实能够增强木糖发酵重组酵母的抗逆能力,提高其发酵性能。该研究成果在木质纤维素替代粮食生产乙醇的产业化进程中具有良好的应用前景。     

产甘油假丝酵母细胞回用对甘油生产的影响

研究了产甘油假丝酵母细胞回用生产甘油的反复分批发酵。实验结果表明：第一级发酵培养基KH2PO4浓度为0.4 g·L-1,回用培养基的最适KH2PO4浓度为0.1 g·L-1,当上一批次发酵液中葡萄糖浓度降至10 g·L-1以下时,酵母细胞不经洗涤即可回用;经过15个批次的反复分批发酵过程,甘油的平均产量、平均得率和平均生产能力分别达到138.69 g·L-1、60.17%和2.31 g·L-1·h-1,分别比第一级发酵结果增加了15.74%、15.48%和39.16%;但回用至第15次时,菌体出现严重衰退,因此回用周期以12～14次为宜。     

ORP调控对马克斯克鲁维酵母发酵纤维素水解液的影响

采用通气的方式进行氧化还原电位(oxidation-reduction potential,ORP)调控,研究在添加抑制物条件下Kluyveromyces marxianus 1727-5利用葡萄糖、木糖以及两者混合体系的发酵性能,在此基础上考察了ORP调控策略对玉米秸秆水解液发酵的影响。研究结果表明:在调控ORP策略下,多种混合抑制物对酵母生长代谢造成的损害得以有效改善,细胞活性高,木糖、葡萄糖代谢速率加快。葡萄糖与木糖共发酵时,ORP调控至-150 mV时,葡萄糖发酵时间缩短近30%,木糖醇浓度由3 g·L~(-1)增加到10 g·L~(-1)。ORP调控策略也同样能有效缓解玉米秸秆水解液中较高浓度的多种抑制物对酵母细胞的胁迫,ORP为-100 mV时,相比于对照组,在保持终点乙醇不变的情况下,葡萄糖的发酵时间缩短了22%;木糖消耗由4.88 g·L~(-1)增至10.27 g·L~(-1),木糖醇得率也由0.20 g·g~(-1)提高至0.48 g·g~(-1)。     

硅橡胶膜生物反应器中乙醇发酵与渗透汽化的耦合

用硅橡胶膜生物反应器(SMBR)实验研究连续发酵-渗透汽化的耦合性能。发酵微生物采用酿酒干酵母,所用碳源为工业级葡萄糖。发酵过程由于产物抑制作用,在乙醇质量浓度达到73 g/L时趋于停滞,而耦合渗透汽化膜后,发酵罐内的乙醇质量浓度降低并维持在40 g/L,使发酵可以连续稳定地进行。在SMBR运行达到稳态后,乙醇的体积产率为4.02 g/(L.h)。发酵液中乙醇质量浓度维持在20~63 g/L,聚二甲基硅氧烷(PDMS)膜的总渗透通量为1 220~800 g/(m2.h),分离因子为5~9.2。与传统发酵和分离相同进料质量分数的乙醇溶液相比,乙醇发酵和渗透汽化在硅橡胶膜生物反应器中能相互耦合并得到强化。与较小规模耦合系统(发酵体积1 L和2 L)比较,性能稳定良好。     

四株酵母乙醇发酵的初步研究

比较了休哈塔假丝酵母NLP21、树干毕赤酵母NLP22、NLP23和NLP31,在30 g/L的木糖和混合糖(葡萄糖15 g/L+木糖15 g/L)发酵培养基上以及在培养基中氮源浓度降低到原来1/2和1/10时的发酵性能。结果表明,在30 g/L木糖发酵培养基上,NLP23和NLP31产乙醇质量浓度最高,分别为(11.14±0.13)和(11.15±0.08) g/L。在15 g/L葡萄糖+15 g/L木糖混合糖发酵培养基上,NLP31产乙醇质量浓度最高,为(10.91±0.12) g/L。当发酵培养基中氮源浓度降低到原来的1/2时,NLP23和NLP31产乙醇能力相当,但后者产木糖醇的量增大;当氮源质量浓度降低到原来的1/10时,NLP23和NLP31产乙醇能力随着发酵轮数的增加,逐渐下降,氮源浓度低,降低了乙醇的产量。     

自产气气提与纤维床反应器耦合发酵生产丁醇

采用发酵产物中的二氧化碳(CO2)和氢气(H2)作为循环气提气源,对丙酮丁醇梭菌(Clostridium acetobutylicum CGMCC 5234)发酵产物进行原位气提,实现丙酮、丁醇和乙醇混合物(ABE)的连续纤维床固定化发酵生产。连续发酵实验进行了12批次共309 h,总溶剂ABE当量浓度为133.3 g·L-1(其中丁醇83.5 g·L-1,丙酮38.4g·L-1,乙醇11.4 g·L-1),葡萄糖消耗率为1.29 g·(L·h)-1,总溶剂ABE产率为0.431 g·(L·h)-1,转化率为0.333 g·g-1,其中丁醇产率为0.270 g·(L·h)-1,转化率为0.209 g·g-1,发酵液中丁醇浓度控制在8~12 g·L-1,显著优于游离发酵的结果。气提提取之后冷凝的ABE溶液出现分层现象,其中丁醇相丁醇浓度高达603.7 g·L-1,极大地减缓后续分离提纯的负担。结果表明,自产气循环气提与纤维床固定化耦合连续发酵生产ABE(特别是丁醇)的工艺具有可行性和竞争力。     

Hydrogels reinforced with nanoclays with improved response rate

Two series of semiinterpenetrating networks (SIPN) based on linear hydrophilic poly(vinyl alcohol) (PVA) and thermo‐responsive poly(N‐isopropylacrylamide) (PNIPA), physically crosslinked with inorganic clay, are presented. The hydrogels with different crosslinking densities were prepared by varying the content of clay from 1 to 6 wt % and contained linear interpenetrant, PVA in the range of 0.5–1.5 wt %. The effect of clay content on swelling/deswelling behavior and phase transition in PNIPA gels, as well as the feasibility of reinforcing the gels with high molecular weight PVA, were analyzed. The thermal response of hydrogels, followed by DSC, confirmed that the insertion of hydrophilic PVA did not have a significant effect on the onset of the volume phase transition temperature, while the response was faster. The equilibrium degree of swelling of SIPNs and PNIPA hydrogels was in the range of 9–79 and decreased with increasing content of clay. The internal morphology and surface wettability of the hydrogels were investigated by scanning electron microscope analysis and contact angle measurements, respectively. The network structural parameters of the PNIPA and SIPN nanocomposites hydrogels, such as the average molecular weight between crosslinks, M_c, and effective crosslinking density, N_e, were determined by dynamic mechanical analysis. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 44535.     

内构件固定床反应器中不同水分煤的热解特性

通过在有无内构件(传热板和中心集气管)固定床反应器中研究不同水分含量煤的热解特性,考察了两反应器中煤料的升温特性、热解产物分布、焦油品质以及气体产物组成和半焦热值。结果表明,内构件可以强化传热和调节热解产物在反应器内的流动,相对无内构件反应器,有内构件反应器的反应时间缩短近一半。在有内构件反应器中,当煤水分增加,导致煤热解反应要求的时间延长,焦油中轻质组分(沸点低于360℃)含量明显升高,焦油收率先增加后降低,热解水和热解气产率升高,而无内构件反应器的热解产物无明显差异。当加热温度900℃时,煤水分从0.41%(本文中无特殊说明的均为质量分数) 增加至11.68%,焦油产率从9.21%增长到10.74%;当煤水分增加到15.93%,焦油产量下降到10.26%。两反应器气体平均组成随水分增加的变化趋势相似,气体热值均随水分增加呈下降趋势。     

Dehydrochlorination of propylene chlorohydrin with sodium hydroxide and with catholyte

Dehydrochlorination of an aqueous solution of propylene chlorohydrin with sodium hydroxide (12·5 and 30% by weight), with catholyte (5 or 10% by weight NaOH + 14% by weight NaCl) and with milk of lime (15% by weight) were compared. The use of catholyte (10% by weight NaOH + 14% by weight NaCl) enables propylene oxide to be obtained in a yield of 94·3%, with 100% conversion of chlorohydrin. The concentration of 1,2-propylene glycol in the waste is 0·07% by weight.     

Enzymatic Assays Coupled with Mass Spectrometry with or without Embedded Liquid Chromatography

This article reviews monitoring strategies for enzymatic assays coupled with mass spectrometric detection. This coupling has already been shown to be helpful in providing versatile and detailed knowledge about enzyme kinetics. Various available publications address two general approaches. 1) The continuous‐flow setup allows real‐time determination of substrate degradation. Simultaneously, resulting product or potential intermediates can be detected. 2) The online coupled continuous‐flow mixing assay allows the direct coupling of an enzymatic assay to chromatographic separation of complex mixtures. The latest efforts in improving the methodology have been made with regard to miniaturization. This is especially advantageous with regard to reducing costly consumption of chemicals. Finally, these developments are applicable for diverse bioanalytical purposes in the realms of pharmaceutical, biotechnological, food, and environmental research.     

Wettability enhancement of polystyrene with electron cyclotron resonance plasma with argon

Polystyrene cell‐culture substrates were treated with argon glow discharge to make their surfaces hydrophilic. The process was novel in that it used a microwave electron cyclotron resonance (ECR) source for polymer surface modification. The substrates were processed at different microwave powers and time periods, and the surface modification was assessed with by measurement of the water contact angle. A decrease in contact angle was observed with increasing microwave power and processing time. Beyond a certain limit of power and duration of exposure, however, surface deterioration occurred. The optimum conditions for making the surfaces hydrophilic without deterioration of the samples were identified. The plasma parameters were assessed by Langmuir probe measurement. Fourier transform infrared spectroscopy with attenuated total reflectance showed evidence for the induction of hydrophilicity on the surface. The surface micromorphology was examined with scanning electron microscopy. The results prove that the ECR glow discharge was an efficient method for enhancing the wettability of the polymer surfaces. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 1618–1623, 2003     

Molecular mechanics calculations on HIV-1 protease with peptide substrates correlate with experimental data

Molecular models of HIV-1 protease and 21 peptide substrateswith single amino acid substitutions at positions from P4 toP3' were built and compared with kinetic measurements. The crystalstructure of HTV-1 protease with a peptidic inhibitor was modifiedto model the peptide substrate Pro-Ala-Val-Ser-Leu-Ala-Met-Thrfor the starting geometry. Models were built of two reactionintermediates, HIV protease with peptide substrate and withits tetrahedral intermediate. The energy minimization used anew algorithm that increased the speed and eliminated a cut-offfor non-bonded interactions. After minimization the models forsubstrate and tetrahedral intermediate both had root mean squaredeviations of 0.48 Å for all atoms of the HIV proteasecompared to the starting crystal structure. Differences in themodel structures and interaction energies for HIV protease withdifferent substrates were analyzed. The calculated interactionenergies for the 21 HIV protease-tetrahedral intermediate modelsgave a correlation coefficient of 0.64 with the kinetic measurements.The eight substrates with changes in the P1 and P1' residuesnext to the scissile bond gave the highest correlation of 0.93,while the 14 substrates with changes in P2-P2' gave a correlationcoefficient of 0.86. The catalytic mechanism and factors influencingthe catalytic efficiency of the different substrates are discussedin relation to the models. The predictive ability of molecularmechanics calculations is discussed in the context of the statisticalmechanics analysis of the differences in free energy.     

Salivary proteins associated with periodontitis in patients with type 2 diabetes mellitus

The objective of this study was to investigate the salivary proteins that are associated with periodontitis in patients with Type 2 diabetes mellitus (T2DM). Volunteers for the study were patients from the Diabetic Unit, University of Malaya Medical Centre, whose periodontal status was determined. The diabetic volunteers were divided into two groups, i.e., patients with periodontitis and those who were periodontally healthy. Saliva samples were collected and treated with 10% TCA/acetone/20 mM DTT to precipitate the proteins, which were then separated using two-dimensional polyacrylamide gel electrophoresis. Gel images were scanned using the GS-800(TM) Calibrated Densitometer. The protein spots were analyzed and expressed in percentage volumes. The percentage volume of each protein spot was subjected to Mann-Whitney statistical analysis using SPSS software and false discovery rate correction. When the expression of the salivary proteins was compared between the T2DM patients with periodontitis with those who were periodontally healthy, seven proteins, including polymeric immunoglobulin receptor, plastin-2, actin related protein 3, leukocyte elastase inhibitor, carbonic anhydrases 6, immunoglobulin J and interleukin-1 receptor antagonist, were found to be differentially expressed (p < 0.01304). This implies that the proteins may have the potential to be used as biomarkers for the prediction of T2DM patients who may be prone to periodontitis.     

复合扰流柱的螺旋内构件反应器内流体的宏观混合

在管式反应器内插入复合扰流柱的螺旋型内构件,采用脉冲示踪法测定了反应器内流体的停留时间分布,研究了器内流体的宏观混合特性,分析了该内构件强化混合的机理. 结果表明,复合扰流柱的螺旋型内构件反应器内的流体流动更接近于活塞流,其轴向扩散模型参数Pe为螺旋型内构件反应器的1.02~1.28倍,为不带内构件反应器的1.35~1.77倍. 复合扰流柱的螺旋型内构件能显著增强器内的二次流动,从而强化了宏观混合效果.     

Improved adhesion of EVAs with different vinyl acetate contents treated with sulphuric acid

Four ethylene vinyl acetate copolymers (EVAs) containing 9, 12, 18 and 20 wt% vinyl acetate (VA) were treated with concentrated sulphuric acid to improve their adhesion to polychloroprene (PCP) adhesive. The tensile strength and Young's modulus of EVAs decreased as the VA content increased, due to the reduction in crystallinity of the polyethylene blocks in the copolymer. The modifications produced in the EVAs by treatment with sulphuric acid were followed using contact angle measurements (water, 25 °C), ATR-IR spectroscopy and scanning electron microscopy (SEM). Adhesive-bond strength was obtained by T-peel tests on treated EVA/polychloroprene adhesive joints. The vinyl acetate content in the EVA affected the extent, but not the nature, of the surface modification produced by treatment with sulphuric acid. The treatment produced both sulfonation and oxidation on the EVA surfaces. The higher the vinyl acetate content in the EVA, the more significant the modifications produced. Increased T-peel strengths of EVA/polychloroprene adhesive + 5 wt% polyisocyanate joints were obtained and a mixed failure (adhesion failure + cohesive failure in the adhesive) was produced. It was found that, to be effective, the treatment of EVAs must be carried out with 96 wt% sulphuric acid.     

Blends of imidized acrylic polymers with SAN copolymers and with PVC

A series of imidized acrylic polymers of varying structural composition generated by reaction of methylamine with poly(methyl methacrylate) were blended with a range of styrene/acrylonitrile or SAN copolymers (0–33% AN) and with poly(vinyl chloride). On the basis of glass transition behavior determined by differential scanning calorimetry, some but not all imidized acrylic structures were found to be miscible with PVC and with SAN copolymers within a limited window of AN levels. Acid functionality in the imidized acrylics appears to hinder their miscibility with SAN rather significantly and with PVC to a lesser extent. Miscible SAN blends showed lower critical solution temperature behavior whereas miscible blends with PVC did not up to the highest attainable temperatures. The composition factors that influence the phase behavior are described and interpreted in terms of possible mechanisms.