单相和两相发酵体系中Methylomonas Z201细胞的生长和环氧丙烷？…

研究了单相和两相发酵体系中甲基单胞菌Ｚ２０１细胞的生长和环氧丙烷的合成。在单相发酵体系中,底物丙烯和产物环氧丙烷抑制细胞生长,水相中环氧丙烷的浓度达到１．３ｍｍｏｌ／Ｌ。在两相发酵体系中,十六烷作为生长底物甲烷以及反应底物丙烯和分子氧的“储器”,减小了丙烯对细胞生长的抑制作用,水相和十六烷相中环氧丙烷的浓度分别达到１．７ｍｍｏｌ／Ｌ和２．６ｍｍｏｌ／Ｌ。同休止细胞相比,单相和两相发酵体系中辅酶ＮＡ     

两相体系中低聚半乳糖的合成

利用乳糖酶转移半乳糖苷活力,以环己烷和乙酸乙酯为有机相主体总蛋白在１５％水分手条件下,得到占总产物３５％以上的低聚半乳糖。在这两相体系中,研究了温度、缓冲液ＰＨ、乳糖浓度、半乳糖和葡萄糖,以及酶的固定化等因素对低地乳糖合成的影响：温度及起始乳糖浓度对低聚糖的影响不明显;加入半乳糖和葡萄糖对低聚糖的合成有一定的影响;以树脂Ｄ３４５固定化乳糖酶作生物催化剂,低聚糖的得率可达６４．７８％。     

两相体系中菌体转化法制备2-苯乙醇的研究

目的:利用酿酒酵母(Saccharomyces cerevisiae CICIMY008 6)菌体在油酸-水两 相体系中转化L-苯丙氨酸生成2-苯乙醇,以期解除产物抑制的同时降低萃取相油酸对转化的 不利影响,提高2-苯乙醇产量.方法:对2-苯乙醇的生成与菌体生长的关系进行考察,以确 定菌体转化法的可行性;通过单因素试验和正交设计试验获得转化培养基最佳配方;对菌体 转化条件进行优化.结果:向装液量为25mL/250mL转化培养基中加入0.6g 酵母湿菌体,30℃ 、100r/min条件下转化,9h加入等体积油酸,催化27h,产物浓度达4.55g/L.结 论:2-苯乙醇的制备可以使用菌体转化法,该法可在一定程度上克服两相转化体系中油酸的毒性影响.     

果蔬废弃物两相厌氧发酵产己酸的效能研究

利用厌氧菌群生物合成己酸被认为是一种非常有潜力的新型废弃物资源化技术,但是其合成效能的提高是目前亟待解决的关键问题。本研究以实际果蔬废弃物为原料,对两相厌氧发酵产己酸的效能进行了研究。首先优化接种比以提高酸化相的水解转化效率;在此基础上通过调控醇酸比和pH以强化产己酸相的发酵效能。结果显示,果蔬废弃物厌氧产酸的最佳接种比为2∶1,此时水解率和酸化率分别可达到98.1%和83.2%,乙酸和丁酸产量分别达到5.4 g/L和3.3 g/L。合理控制醇酸比和pH对提高产己酸相的发酵效能非常关键。当醇酸比和pH控制为4∶1和7.5时,己酸生成量可达14.9 g/L,约占液相总COD的80.84%;而低醇酸比和低pH易造成丁酸的累积,从而降低了己酸产量。己酸发酵过程属于非生长偶联型,己酸菌(Clostridium kluyveri)指数增长期伴随着丁酸的生成,而己酸合成主要发生在生长中后期。此外,己酸菌对于pH变化较为敏感,适当提高pH有助于减轻有机酸毒性,提高生物量;但是碱性环境会严重抑制己酸菌的生长繁殖。研究表明,通过分别对酸化相和产己酸相进行优化和调控,两相发酵策略更有利于提高己酸合成效能。     

微水-有机溶剂两相体系中消旋萘普生的酶法拆分

研究了微水-有机溶剂两相体系中固定化脂肪酶催化的萘普生甲酯的立体选择性水解反应。固定化酶活性受载体极性、水含量、有机溶剂的logP值、产物抑制的影响,据此构建了一种可以连续拆分产生(S)(+)萘普生的微水-有机溶剂两相体系。反应在一个具有回路的连续流搅拌反应器中进行,反应器中添加有采用吸附法固定化的脂肪酶,载体为一种弱极性的合成载体,水相连同固定化酶颗粒一起永久保持在反应器中,有机流动相带入底物,带出产物。固定化酶在该50mL反应器中30℃连续操作60d,仅损失活性25%,产生(S)(+)萘普生900mg,产物对映体过量值(ee_p)为95%。     

水-有机溶剂两相体系中甲基单孢菌Z201细胞催化丙烯环氧化的初步研究

Laabe于1987年提出了生物催化剂工程(Biocatalyst engineering)和介质工程(Medium engineering)的概念^[1]。有机相生物催化中溶剂的选择也是介质工程的内容之一。纯酶在有机相中的催化作用已有大量报道^[2],但对完整细胞研究甚步。本文以甲基单胞菌(Methylomonas Z201)完整细胞为生物催化剂．丙烯环氧化为指标反应．研究有机溶剂对活性的影响并对催化活性——溶剂疏水性进行了相关性分析。研究了水一十六烷两相体系中十六烷含量和搅拌速度对丙烯环氧化速度的影响和细胞的操作稳定性。     

水-有机溶剂两相体系中甲基单胞菌Z201催化丙烯环氧化的初步研究

Lanne于1987年提出了生物催化剂工程（Biocatalyst engimeering）和介质工程（Medium enineering）的概念^[1].有机相生物催化中溶剂的选择也是介质工程的内容之一。纯酶在有机相中的催化作用已有大量报道^[2],但对完整细胞研究甚少。本文以甲基单胞菌（Methylomonos）Z201完整细胞为生物催化剂,丙烯环氧化为指标反应,研究有机溶剂对活性的影响并对催化活性-溶剂疏水性进行了相关性分析。研究了水-十六烷两相体系中十六烷含量和搅拦速度对丙烯环氧化速度的影响和细胞的操作稳定性。     

果糖和麦芽糖作为有氧碳源对大肠杆菌NZN111两阶段发酵中厌氧发酵的影响

为了了解磷酸转移酶转运系统 (PTS) 依赖和非PTS依赖代谢的糖类对大肠杆菌生产琥珀酸的影响,进行了两阶段培养,有氧阶段采用PTS依赖型的果糖或非PTS依赖型的麦芽糖作为丙酮酸甲酸裂解酶 (PFL) 和乳酸脱氢酶 (LDH) 双突变株NZN111的碳源,研究其对NZN111厌氧阶段代谢葡萄糖的影响。5 L罐发酵结果表明,以果糖和麦芽糖为碳源有氧培养的细胞恢复了在厌氧条件下快速代谢葡萄糖的能力,琥珀酸和丙酮酸成为主要代谢产物,最终琥珀酸得率分别为0.84和0.75 mol/mol,丙酮酸得率分别达到了0.65和0.83 mol/mol,琥珀酸和丙酮酸终浓度比分别为1.73∶1和1.21∶1。果糖和麦芽糖培养的NZN111与葡萄糖培养的菌体代谢的明显差异推测是cyclic AMP (cAMP) 依赖型和非cAMP依赖型的分解代谢物阻遏调控这两种机制共同作用的结果。     

糙皮侧耳不同生长时期发酵料中微生物和代谢物的变化

为了解糙皮侧耳栽培期间发酵料中的微生物及代谢产物变化,分别采用宏基因组测序、非靶向代谢组学技术对糙皮侧耳5个生长时期发酵料中微生物及代谢物进行分析.结果 表明:糙皮侧耳生长发育过程中,发酵料中的Bacillus EGD-AK10、Cellulosinicrobium aquatile、Cellulosinicrobiu...     

花生四烯酸高产突变株的选育及其发酵调控

以拉曼被孢霉（Mortierellaramanniana）SM541为原始菌株,经过紫外线复合氯化锂诱变处理,得到突变株SM541－9,其生物量由126g／L提高到28．8g／L,油脂含量由5．8g／L提高到15.7g／L。花生四烯酸含量由321mg／L增加到623mg／L。传代实验表明,SM541－9具有良好的遗传稳定性,以葡萄糖为碳源,硝酸钾为氮源的最适培养条件下,10L罐发酵,生物量和花生四烯酸含量较原始菌株分别提高了45．2％和109.5％。在液体发酵过程中,菌丝老化、增大溶氧量、间歇流加葡萄糖都有利于花生四烯酸的合成。     

2—KLG产生菌混合发酵特性及最佳混生模式的研究

氧化葡萄糖酸杆菌合成的2－KLG对巨大芽孢杆菌的生长繁殖具有明显的抑制作用,可缩短其生长周期。发酵体系中巨大芽孢杆菌的存在是氧化葡萄糖酸杆菌的生长繁殖和合成2－KLG所必需的,发酵过程中巨大芽孢杆菌裂解所释放的活性物质可能是刺激氧化葡萄糖酸杆菌合成2－KLG的主要原因。二菌混合发酵需在适宜的混生模式下才可达到最佳效果。     

EVALUATION OF BIOETHANOL PRODUCTION FROM CAROB PODS BY Zymomonas mobilis AND Saccharomyces cerevisiae IN SOLID SUBMERGED FERMENTATION

Bioethanol production from carob pods has attracted many researchers due to its high sugar content. Both Zymomonas mobilis and Saccharomyces cerevisiae have been used previously for this purpose in submerged and solid-state fermentation. Since extraction of sugars from the carob pod particles is a costly process, solid-state and solid submerged fermentations, which do not require the sugar extraction step, may be economical processes for bioethanol production. The aim of this study is to evaluate the bioethanol production in solid submerged fermentation from carob pods. The maximum ethanol production of 0.42 g g^?1 initial sugar was obtained for Z. mobilis at 30°C, initial pH 5.3, and inoculum size of 5% v/v, 9 g carob powder per 50 mL of culture media, agitation rate 0 rpm, and fermentation time of 40 hr. The maximum ethanol production for S. cerevisiae was 0.40 g g^?1 initial sugar under the same condition. The results obtained in this research are comparable to those of Z. mobilis and S. cerevisiae performance in other culture mediums from various agricultural sources. Accordingly, solid submerged fermentation has a potential to be an economical process for bioethanol production from carob pods.     

酪氨酸对表面灌流系统中大鼠离体大,小黄体细胞孕酮生成的影响

参照我室Ｗａｎｇ Ｘｉａｏ－Ｎｉｎｇ的黄体细胞制备方法，制成大鼠黄体细胞悬浮液，经蔗糖密度梯度离心分离大，小黄体细胞，经６０ｍｉｎ预培灌后，每１０ｍｉｎ收集一次培灌液，用ＲＩＡ测定其中孕酮含量，实验结果表明，在培灌系统中，大黄体细胞的基础孕酮分泌量明显高于小黄体细胞，但小黄体地ｈＣＧ的敏感性强于大黄体细胞，不同剂量的酪氨酸对ｈＣＧ致大，小黄体细胞孕酮生成有不同程度的抑制，对小黄体细胞的抑制作用较为     

ANTISENSE INHIBITION OF THE 27 kDa HEAT SHOCK PROTEIN PRODUCTION AFFECTS GROWTH RATE AND CYTOSKELETAL ORGANIZATION IN MCF-7 CELLS

MCF-7 cells were co-transfected with the human HSP27 antisense cDNA and the neomycin resistance gene, included in the constitutive expression vector pSVL, and the phenotypical changes associated with decreased expression of the HSP27 protein were analysed. Three out of 10 neomycin-resistant clones obtained proliferated normally and showed a normal HSP27 content (Western blot). The seven other clones (designated as αHSP27 clones) were characterized by a dramatic growth inhibition associated with alterations in cellular morphology. Cells became progressively hypertrophied, exhibited lamellar protrusions and tended to lose contact with each other. They also acquired characteristics of secretory cells, namely the presence of numerous refractile granules and secretory canaliculi. Among the αHSP27 clones, two were immunocytochemically analysed for HSP27 content. Both clones were immunonegative for HSP27, contrary to parental cells and neo transfectants. Actin immunostaining in one of these HSP27 negative clones revealed that microfilament organization changed from diffuse to punctate distribution. Our data support the current concept of a role for HSP27 in cell growth and differentiation and further suggests that this might occur through a control on actin polymerization-depolymerization.     

IN SITU SEPARATION OF ETHANOL WITH AQUEOUS TWO-PHASE SYSTEM AND ASSESSMENT OF KLa FOR YEAST GROWTH IN BATCH CULTIVATION

In the fermentation process, the separation of product and its purification is the most difficult and exigent task in the ground of biochemical engineering. Another major problem that is encountered in the fermentation is product inhibition, which leads to low conversion and low productivities. Extractive fermentation is a technique that helps in the in situ removal of product and better performance of the fermentation. An aqueous two-phase system was employed for in situ ethanol separation since the technique was biofriendly to the Saccharomyces cerevisiae and the ethanol produced. The two-phase system was obtained with polyethylene glycol 4000 (PEG 4000) and ammonium sulfate in water above critical concentrations, with the desire that the ethanol moves to the top phase while cells rest at the bottom. The overall mass transfer coefficient (K_La) was also estimated for the yeast growth at different rpm. The concentration and yield of ethanol were determined for conventional fermentation to be around 81.3% and for extractive fermentation around 87.5% at the end of the fermentation. Based on observation of both processes, extractive fermentation was found to be the best.