固定化紫草细胞生产紫草宁衍生物

用海藻胶包埋紫草细胞,在紫草色素生产培养基M_9中,常温、黑暗下培养不同时间,收集培养液并提取色素,进行紫外—可见全波长分光光度扫描和TLC分析。结果表明:固定化紫草细胞可连续分泌紫草色素,其主要成分与天然成分基本相同,产量已达到一定水平。此外对海藻胶包埋条件、固定化珠粒的稳定性以及1L固定化植物细胞反应器生产紫草色素工艺进行了讨论。     

产物释放促进剂对固定化紫草细胞生产紫草色素的影响

研究了多种产物释放促进剂对海藻胶包埋的紫草细胞生产紫草色素的效果。结果表明,在色素生产培养基中加入合适的产物释放促进剂可显著地刺激紫草色素的分泌及合成,并且该种处理方式的作用效果明显优于细胞透性预处理方式的效果。将固定化紫草细胞培养在质量分数为０．０８％、０．２４％Ｔｗｅｅｎ８０和０．２％Ｔｗｅｅｎ２０培养基中,培养１２ｄ紫草色素产量分别为每克干细胞含４２．２０、６２．５２和３８．７３ｍｇ,比对照分别提高２．８２、４．６６和２．５１倍。在质量分数为０．２％Ｔｗｅｅｎ２０培养基中培养３５ｄ时,细胞合成色素总产量达每克干细胞含１８１．５ｍｇ,比对照提高４．２倍。     

聚氨酯泡沫原位提取培养紫草细胞生产紫草色素

在紫草细胞培养过程中影响紫草色素进一步合成的重要因素之一是色素在细胞内大量积累，为促进紫草色素向胞外分泌，建立了一种新的培养体系，即以聚氨酯泡沫作为色素吸附介质，加入到细胞培养液中原位吸附提取培养紫草细胞生产紫草色素。结果表明：聚氨酯泡沫的加入明显促进紫草色素的合成及分泌，细胞合成的９５％（质量分数，下同）以上的色素被聚氨酯泡沫吸咐分离，同时也发现在培养早期（０４ｄ）加入适量的（９ｇ／Ｌ）聚氨酯泡沫可使色素产量提高３０８倍。采用丙酮和乙醇洗脱吸附的紫草色素具有优良的解吸效果。另外，试验还比较了原位提取培养体系与非原位提取培养体系细胞对蔗糖利用情况以及ｐＨ值变化情况。     

紫草细胞悬浮培养的动力学

通过对紫草细胞进行悬浮培养,测定了培养基中碳源、氮源、磷酸盐等无机元素的变化;并分析了细胞结构性组分、细胞呼吸以及次生代谢产物紫草宁的合成情况,探索研究了细胞培养的内在规律,为建立结构化动力学模型奠定了基础。     

多糖对紫草细胞生产紫草色素的影响

在紫草色素生产培养基中添加琼脂糖、人参多糖、海藻多糖、黄原胶、卡拉胶和淀粉等６种多糖类物质，接入紫草细胞后置于２５℃黑暗条件下振荡培养２０ｄ，结果表明：试验中所选用的６种多糖类物质对紫草色素的产生均具有一定的刺激作用，其中以０．０５％琼脂糖的作用效果为最佳，紫草色素含量为２０．７７％，比对照提高４６．８９％。此外试验亦对琼脂糖对紫草色素生产及分泌特征、培养液中蔗糖浓度变化以及ｐＨ值变化情况进行了研究。     

细胞培养生产的紫草色素与天然紫草色素理化性质比较

对细胞培养生产的紫草色素与天然紫草色素进行了理化性质比较研究，结果表明，两者的耐热性、耐氧化性及不同ｐＨ值条件下颜色的变化无明显差异，而耐还原性则具显著差异。此外，还对细胞培养生产的紫草色素和天然紫草色素的组成成份进行了ＴＬＣ和ＨＰＬＣ分析，发现两者组成成份基本一致，只是相对含量具一定差异．     

半连续悬浮细胞及固定化细胞生产紫草素的研究

采用半连续悬浮细胞及固定化细胞生产紫草素,结果表明:前者可明显提高单位细胞的紫草素产量,但在15天后其产量明显下降;后者可在75天内连续稳定地生产并分泌紫草素。采用六种化学试剂,以不同浓度处理固定化细胞,发现只要选择合适的试利与浓度即可改变细胞的透性而不影响其活性,并且产生的紫草素可不断地向胞外分泌,产量可提高2.4倍。应用紫外——可见全波长扫描、TLC 和 HPLC 分析,发现固定化细胞生产的紫草素与天然的紫草素成分基本相同,只是各单体成分的相对含量有一定的差异。     

一步法培养紫草细胞生产紫草宁工艺与机理研究

本文提出一步法培养生产紫草宁新工艺，将细胞生产和产物合成两阶段合二为一，免去了原两步法培养中细胞生长阶段后必须充分过滤、洗涤等操作．该工艺操作简单、生产率高、细胞可重复利用．培养的紫草于占细胞干重已达17．9％，色素产量最高达2．3g／L.一步法培养的细胞重复使用试验表明，再生细胞的色素含量与原细胞的色素含量相近，一步培养末期进行半连续培养是可行的．在固体培养中，以一步培养基取代M－9培养基，接种量小，收获时紫草宁占细胞干重可达IS．7％，色素产量为2．7g从．本文对紫草合成机理进行了分析，提出了新疆紫草细胞…     

吸附树脂对紫草细胞培养生产紫草色素的原位提取

紫草（Ｌｉｔｈｏｓｐｅｒｍｕｍｅｒｙｔｈｒｏｒｈｉｚｏｎ）细胞在Ｍ－９培养基中培养时产生红色萘醌类化合物。实验表明，若在培养液中加入ＨＤ－１树脂，使色素和细胞产量下降，而Ｒ－Ａ树脂的加入却促进色素的合成和分泌，培养４ｄ的悬浮细胞中加入３０ｇ／ＬＲ－Ａ树脂，可使色素产量提高２倍。此外，还比较了原位提取和非原位提取体系细胞对碳源蔗糖的利用。     

超声波对新疆紫草悬浮培养细胞生长和紫草素合成的影响

探讨了超声波功率、超声波处理时间对新疆紫草细胞生长和紫草素合成的影响. 研究结果表明,超声波对悬浮培养的新疆紫草细胞生长有促进作用,低功率长时间或高功率短时间的超声处理对细胞生长比较有利. 在优化条件下(200 W超声处理1 min),培养结束时的生物量比对照提高61%. 超声波也可以提高新疆紫草悬浮培养细胞的紫草素含量和产量,接种后即进行超声波处理,超声波功率密度为39.9 mW/cm3、超声时间为3 min时,细胞紫草素含量和产量最高,达到2.72%和294 mg/L,分别比对照组提高了64%和135%. 超声波是通过提高细胞苯丙氨酸解氨酶的活力来强化紫草素的生物合成途径.     

包埋紫草细胞的海藻酸钙凝胶扩散性能

用非稳态法测定了充分搅拌条件下在未包埋紫草细胞和包埋紫草细胞的海藻酸钙凝胶珠粒体系内蔗糖溶液浓度的瞬间变化情况。结果表明，海藻酸钠浓度、氯化钙浓度和紫草细胞包埋量对蔗糖扩散有明显影响，有效扩散系数Ｄｅ随着海藻酸钠浓度、氯化钙浓度和紫草细胞包埋量的提高而下降．     

聚氨酯泡沫固定化重组枯草杆菌三段培养生产青霉素酰化酶

引　言青霉素G酰化酶 (penicillinGacylase ,EC3 5 1 11,PGA)是 β 内酰胺类抗生素工业中的关键酶之一 .目前主要采用大肠杆菌和巨大芽孢杆菌进行工业化生产 ,但由于产酶水平较低、变温发酵和需要苯乙酸诱导等缺点 ,国内外研究者尝试采用各种基因重组技术 ,以期大幅度提高产酶     

羧基修饰的超顺磁纳米粒子直接固定化罗伊氏乳杆菌

利用共沉淀法结合高锰酸钾氧化制备所得表面羧基修饰的超顺磁性纳米粒子吸附于罗伊氏乳酸杆菌表面,在磁场协助下实现细胞的固定化。吸附机理分析表明小尺寸相互作用和静电相互作用是磁性纳米粒子与细胞之间的主要作用。分别考察了菌体/磁性粒子质量比、pH、温度、时间等对固定化罗伊氏乳酸杆菌的影响,确定最佳固定条件为菌体与磁性纳米粒子相对质量比为2.25,在pH=3、温度25℃的条件下固定化0.5 h,可实现91%的细胞固定化。最后,对固定化后的细胞进行再培养,与游离细胞相比,两者表现出类似的代谢特征,证实细胞经固定化后仍具有活性。因此,羧基修饰的超顺磁性纳米粒子可成功用于细胞固定化,在不影响细胞活性的情况下,通过磁分离实现细胞的重复利用。     

Studies on the structural features of sodium alginate entrapped Kluyveromyces marxianus NCYC179 cells used for alcohol production from whey permeate

Alginate-entrapped living Kluyveromyces marxianus NCYC179 cells were successfully used in alcohol production from whey permeate. Using 10% whey lactose as substrate, an alcohol production efficiency of 81.5–84.9% and 84.0–88.2% with cell viability maintenance between 82.8–84.3% and 81.0–84.3% of the immobilised cells during the monitoring periods, in batch (six repeat runs) and continuous (23 days of experimenting time) systems, respectively. The immobilised samples remained stable during the course of fermentation with no leakage of K. marxianus cells into the surrounding medium in the case of the batch system. However, slight leakage after 10 days of continuous run in the continuous process was recorded. The scanning electron microscopic studies of the beads containing entrapped yeast cells and sections or crushed samples of the beads carried out at the start and termination of fermentation in both the processes (batch and continuous) revealed that the immobilisation procedure does not influence any morphological change in the entrapped cells during the prolonged periods of fermentation. Moreover, the gel matrix structure was also found to be unaffected by the fermentation conditions employed during the course of present studies.     

非水相体系中海藻酸钠-壳聚糖微胶囊粒径及强度性能

固定化技术是提高生物催化剂在非水相中活性和稳定性的一种很有工业应用潜力的手段。采用乳化-内部凝胶化与络合反应制备海藻酸钠-壳聚糖(AC)微胶囊作为固定化载体,筛选5种有机溶剂构建了培养基-有机溶剂两相体系,模拟酵母细胞培养条件,将AC微胶囊在培养基-有机溶剂两相体系中振荡48 h,5种两相体系对AC微胶囊形态没有明显影响;在培养基-癸二酸二丁酯两相体系中,当壳聚糖分子量在40000～100000、成膜反应时间在2～5 min范围内变化时,分子量小(40000)和成膜时间长(5 min)的AC微胶囊粒径稳定,破碎率较低、机械强度较大,适于用作进一步非水相细胞催化的固定化载体。两相体系中AC微囊化酵母细胞活性保持良好,能实现生物转化生产。     

Use of polyelectrolyte complex for immobilization of microorganisms

Cells from Escherichia coli (IAM 12119) were immobilized with the polyion complex of trimethylammonium glycol chitosan iodide (TGCI) and potassium poly(vinyl alcohol) sulfate (KPVS). The immobilization was carried out at pH 8 by mixing TGCI with the cell suspension, followed by addition of KPVS. The immobilized cells were characterized by investigating the glucose oxidizing activity. The results obtained indicated that the glucose consumption with immobilized cells is due not only to the cells released from the complex support but also to the entrapped cells which are grown in the complex; therefore, the cells entrapped in the complex have the glucose oxidizing activity. The physicochemical studies on the immobilization mechanism showed that cells are immobilized via two stages: the aggregation of cells with TGCI and the entrapment of the aggregates in the TGCI–KPVS complex. In the aggregation process, a part of TGCI which is added to cell suspension adsorbs on the cells and the other remains in the suspension. In the entrapment process, the remainder forms the polyion complex with KPVS added and the aggregated cells are coprecipitated with the complex.     

Polyurethane Foam Rafts Supported In Vitro Cultures of Rindera graeca Roots for Enhanced Production of Rinderol,Potent Proapoptotic Naphthoquinone Compound

Unique phytochemical profile of plants belonging to Boraginaceae family provides a prolific resource of lipophilic pigments from the group of naphthoquinone derivatives. To overcome low compound content, the major obstacle of plant-based production, immobilization of Rindera graeca roots in in vitro cultures was implemented for efficient production of rinderol, novel furanonaphthoquinone derivative with anticancer properties. Chromatographic procedures revealed rinderol presence in extracts of all investigated root lines, derived both from root biomass and post-culture medium. Unexpectedly, in the second stage of the experiment, rinderol production was ceased in control, unmodified culture systems. On the contrary, roots immobilized on PUF rafts uniformly and stably produced rinderol, and its highest amount was noted for transformed root lines after 42 days of cultivation (222.98 ± 10.47 µg/flask). PUF occurred to be the main place of compound accumulation. Moreover, investigation of rinderol biological activity revealed its fast-acting cell death induction in HeLa cervical cancer cells at relatively low concentrations. Presented results revealed successful application of R. graeca roots immobilization on PUF rafts for production and in situ product removal of rinderol, novel lipophilic furanonaphthoquinone with suggested proapoptotic activity.