几种营养物质对烟草细胞生长和CoQ_(10)含量的影响

研究了蔗糖、KH2PO4、NH 4/NO3比对烟草细胞生长和CoQ10含量的影响,结果表明,当蔗糖浓度为30g/L时,CoQ10总量最高,此时细胞产量、CoQ10含量和总量分别为198g/L、4147μg/g(dwt)、8212μg/L。在上述蔗糖浓度下,当KH2PO4起始浓度为340mg/L、NH 4/NO-3为12时,细胞产量、CoQ10含量和总量分别最高,高于此比例时有利于CoQ10形成,但不利于细胞生长。     

磁胁迫对烟草细胞生长和CoQ10含量的影响

用0.4 T恒定磁场处理烟草细胞0.5～1.5 h后,细胞活力和存活率可受到一定的影响,但处理间差异不显著.处理0.5～1.0h的细胞分裂指数和细胞生物量显著提高,分别提高15.4%和49.6%,细胞干重分别提高18.4%和42.3%,但CoQ10的生物合成和积累受抑.处理1.5 h的细胞分裂和细胞生长受抑,但CoQ10的生物合成和积累得到促进,CoQ10含量提高25.2%.     

生长调节剂对石刁柏悬浮培养细胞的生长和多糖含量的影响

选取在MS十2,4DI．0ms／L（单位下同）＋6－BA0．2培养基上,每30～40d继代1次的一年生长石刁柏（As～（nxv）zllaLLi）愈伤组织无性系,于26士IC下进行无菌悬浮培养,培养基为MS＋2,4D0·4＋6－BA0．4,每4d换液1次,8～12d继代1次,3个月后用于实验。测定生长（60C下供至恒重）和多糖（0．15g干培养物加3ml水,超声波处理回～2min后匀浆,于90C水浴中置lh,离心,上清液加2倍体积酒精沉淀过夜,取沉淀加入1．5ml水）。多糖测定按苯酚硫酸法（张惟杰．复合多糖生化研究技术．上海：上海科学技术出版社,1987．6）。结果表明：回…     

细胞外钙调素对烟草悬浮培养细胞蛋白质磷酸化作用的影响

用液体闪烁计数法研究了细胞外钙调素对烟草悬浮培养细胞质蛋白质磷酸化的作用。结果表明烟草细胞细胞质蛋白质磷酸化活性在细胞培养过程中逐渐增加,达到最高峰后又开始下降。在细胞质蛋白质磷酸化强度高峰时,加入抗CaM血清后,细胞质蛋白质磷酸化活性受到了部分抑制。加抗CaM血清后再补加CaM能够部分解除抗CaM血清对细胞质部分与细胞核部分蛋白质磷酸化的抑制作用。外加纯化钙调素可以引起烟草悬浮培养细胞细胞质蛋白质磷酸化的活性增强,并且这种增强作用具有时间（高峰为70min）与剂量（最适为CaM10-7mmol/L）依赖性。CaM引起的细胞质蛋白质磷酸化变化与红光所引起的细胞质蛋白质磷酸化变化在时间进程上是不相同的。     

影响高山红景天细胞悬浮培养中细胞生长和红景天苷积累的几个…

高山红景天细胞悬浮培养过程中３ｍｇ·Ｌ＾－１６－ＢＡ,０．３ｍｇ·Ｌ＾－１ＮＡＡ,６０ｍｍｏｌ·Ｌ＾－１氮源、０．５￣１．２５ｍｍｏｌ·Ｌ－１ＫＨ２ＰＯ４和２０ｍｇ·Ｌ＾－１蛋白胨,较适合于细胞生长和红景天苷的积累。在适宜的接种量下细胞培养２４ｄ后,生物量达１４．０４ｇ·Ｌ＾－１,干细胞中红景天苷含量为５．６６ｍｇ·ｇ＾－１。     

细胞外钙调素对烟草悬浮培养细胞蛋白质磷酸化作用的影响

用液体闪烁计数法研究了细胞外钙调素对烟草悬浮培养细胞质蛋白质磷酸化的作用。结果表明：烟草细胞细胞质蛋白质磷酸化活性在细胞培养过程中逐渐增加,达到最高峰后又开始下降。在细胞质蛋白质磷酸化强度高峰时,加入抗CaM血清后,细胞质蛋白质磷酸化活性受到了部分抑制。加抗CaM血清后再补加CaM能够部分解除抗CaM血清对细胞质部分与细胞核部分蛋白质磷酸化的抑制作用。外加纯化钙调素可以引起烟草悬浮培养细胞细胞质蛋白质磷酸化的活性增强,并且这种增强作用具有时间（高峰为70min）与剂量（最适为CaM10^-7mmol/L）依赖性。CaM引起的细胞质蛋白质磷酸化变化与红光所引起的细胞质蛋白质磷酸化变化在时间进程上是不相同的。     

4种红豆杉培养细胞的生长和紫杉醇含量的比较(简报)

南方红豆杉、云南红豆杉、欧洲红豆杉和东北红豆杉细胞生长在45d的培养过程中基本至S型,在30d时生长速率最大,以后逐渐下降。随着细胞进入快速生长期,培养细胞中的紫杉醇含量也达到最大值,其中云南红豆杉含量最高（0.019％）,南方红豆杉含量居中（0.009％）,而欧洲红豆村和东北红豆杉含量最低（0.004％和0．006％）。     

几种生理因子对印楝细胞悬浮培养生长的影响

通过研究不同生理因子对印楝悬浮细胞生长的影响 ,建立了一种快速生长的印楝细胞悬浮培养体系。结果表明 ,在添加了 6 BA 0 .8mg/L、NAA 0 .8mg/L、蔗糖 3 0 g/L且初始 pH值为 5 .8的MS培养液中 ,以 60g/L的接种量进行印楝细胞的悬浮培养 ,细胞生长速率可达 4.49g/L·d。     

一种改进的烟草悬浮细胞继代方法

在植物细胞悬浮培养过程中,对影响细胞系均一性和质量的细胞团块形成的克服方式一般有2种。一是分级过滤继代,即用不同目数的细胞筛除去大的细胞团块和细胞碎片,将小细胞团和单细胞转移到新培养基中作继代培养。此方法操作     

微生物CoQ合成途径及CoQ10生产菌株的分子生物学改造

CoQ10具有呼吸链电子传递者、抗氧化性、调控基因表达等多种生理生化功能,目前不仅用作药物也用作食品添加剂。微生物发酵法是目前生产CoQ10最有效的方法。本文就有关微生物CoQ合成途径及基于CoQ合成途径的CoQ10生产菌株分子生物学改造的策略与研究进展进行了综述和展望。     

Effects of Nutritional Factors on the Formation of Ubiquinone by Tobacco Plant Cells in Suspension Culture

The ubiquinone (UQ) content of BY–2 cells on surface culture was examined to compare with that in suspension culture. The UQ content on surface culture was a little lower than that in suspension culture, but the pattern of the time-course of the UQ formation on surface culture was similar.

The changes of UQ content in BY–2 cells during autolysis were also examined. UQ in the cells subjected to autolysis was not rapidly metabolized nor excreted into the medium.

In order to obtain basic information for UQ formation by BY–2 cells in suspension culture, the cultural conditions, especially nutritional ones were investigated. Addition of 2,4-D was remarkably effective on UQ formation and a higher UQ content was observed with a higher 2,4-D concentration. Sucrose and glucose concentrations in the original medium were also influential factors. The UQ content tends to increase with the decrease of the sugar content. Precursors of UQ, amino acids, vitamins and organic acids were not effective on the UQ formation.     

Investigation of the Effects of Squalene and Squalene Epoxides on the Homeostasis of Coenzyme Q10 in Rats by UPLC‐Orbitrap MS

Squalene has been used as a dietary supplement for a long history due to its potential cancer‐preventive function. However, the mechanism has not been investigated in detail yet. Therefore, the aim of this study is to see if the plasma coenzyme Q10 (CoQ10) level will be altered by gavage of squalene and oxidosqualenes to rats. In the present work, a sensitive and simple high‐performance analytical method based on ultra‐high‐performance liquid chromatography coupled with an Orbitrap mass spectrometry (UPLC‐Orbitrap‐MS) was developed for the quantification of CoQ10 in rat plasma. Coenzyme Q9 (CoQ9) was employed as the internal standard. CoQ10 was determined after acetonitrile‐mediated plasma protein precipitation using UPLC‐Orbitrap‐MS in negative ion mode. Intragastric administration of squalene and the two squalene epoxides into rats once daily for several days elevated the level of CoQ10 in their plasma, but there was no significant difference between high‐dose (286 mg/kg) and low‐dose (143 mg/kg) groups. Intragastric administration of squalene once a day for 5 consecutive days and oxidosqualenes once a day for 3 consecutive days is necessary for reaching the steady‐state level of CoQ10. Our present findings indicate that squalene and oxidosqualenes may be useful for stimulating the synthesis of CoQ10 in rats.     

Assembly of Coenzyme Q10 nanostructure resembling nascent discoidal high density lipoprotein particle

There are tremendous drug candidates that suffer from insolubility in water. In the present study, it is shown that Coenzyme Q10 (CoQ10), a model water-insoluble compound, can be nanoparticulated into a water-soluble form using apolipoprotein A-I (apoA-I). Similar to the way that apoA-I forms nascent discoidal high density lipoprotein (ndHDL) particles by bordering acyl chain tails of phospholipids, CoQ10 could be enclosed into the circle of a disk made of apoA-Is. The resulting nanostructure of CoQ10 and apoA-I was water-soluble with a size of ∼12 nm in diameter and was physically more robust than liposome. We expect that the strategy suggested in this study can be exploited to assemble nano-sized, water-soluble structures of various water-insoluble drug candidates.     

辅酶Q10/两亲性木聚糖纳米悬浮剂的制备以及生物利用度

研究一种新型共聚物负载辅酶Q10形成纳米悬浮剂能够增加CoQ10的水溶性,并且提高其口服生物利用度。本研究以槲皮素—木聚糖（QT-Xylan）共聚物偶联为基础进行合成,采用高剪切均质法进一步包载辅酶Q10,形成了一种新型载药纳米悬浮剂。采用单因素实验设计,并以粒径大小作为单因素实验的考察条件,影响其粒径大小的因素包括高压均质压力、高压均质次数、共聚物浓度、共聚物与CoQ10的质量比4个因素,并进行一系列体外实验评价。当均质压力为60 MPa,均质次数为7次,共聚物浓度为1 mg·mL^-1,共聚物与CoQ10的质量比为1∶1,是纳米悬浮剂的最佳制备工艺,此时粒径大小为166.7 nm。在最佳工艺条件下,在体外溶出实验中,包载CoQ10纳米悬浮剂的体外溶出率在人工胃液（SGF）和人工肠液（SIF）中分别是CoQ10原药的1.89和1.48倍。在体内生物利用度实验中,分别对大鼠灌胃CoQ10原药与载药纳米悬浮剂后,检测不同时间点的血药浓度,考察药物在大鼠体内的吸收和代谢情况,负载CoQ10的纳米悬浮剂在大鼠体内的血药浓度明显高于CoQ10原药,生物利用度提高为CoQ10原药的2.64倍。     

Coenzyme Q10 production in plants: current status and future prospects

Coenzyme Q10 (CoQ10) or Ubiquinone10 (UQ10), an isoprenylated benzoquinone, is well-known for its role as an electron carrier in aerobic respiration. It is a sole representative of lipid soluble antioxidant that is synthesized in our body. In recent years, it has been found to be associated with a range of patho-physiological conditions and its oral administration has also reported to be of therapeutic value in a wide spectrum of chronic diseases. Additionally, as an antioxidant, it has been widely used as an ingredient in dietary supplements, neutraceuticals, and functional foods as well as in anti-aging creams. Since its limited dietary uptake and decrease in its endogenous synthesis in the body with age and under various diseases states warrants its adequate supply from an external source. To meet its growing demand for pharmaceutical, cosmetic and food industries, there is a great interest in the commercial production of CoQ10. Various synthetic and fermentation of microbial natural producers and their mutated strains have been developed for its commercial production. Although, microbial production is the major industrial source of CoQ10 but due to low yield and high production cost, other cost-effective and alternative sources need to be explored. Plants, being photosynthetic, producing high biomass and the engineering of pathways for producing CoQ10 directly in food crops will eliminate the additional step for purification and thus could be used as an ideal and cost-effective alternative to chemical synthesis and microbial production of CoQ10. A better understanding of CoQ10 biosynthetic enzymes and their regulation in model systems like E. coli and yeast has led to the use of metabolic engineering to enhance CoQ10 production not only in microbes but also in plants. The plant-based CoQ10 production has emerged as a cost-effective and environment-friendly approach capable of supplying CoQ10 in ample amounts. The current strategies, progress and constraints of CoQ10 production in plants are discussed in this review.     

Effects of Inorganic Nitrogen Sources and Physical Factors on the Formation of Ubiquinone by Tobacco Plant Cells in Suspension Culture

In order to obtain basic information on ubiquinone (UQ) formation by BY-2 cells in suspension culture, effects of inorganic phosphate and nitrogen sources and such physical factors as initial pH, light irradiation, shaking condition and temperature were investigated. The concentration of phosphate and nitrogen sources had no marked effect, but the ratio of ammonium nitrogen to nitrate nitrogen was significantly effective on UQ formation. The UQ content in BY-2 cells tends to increase at higher ratios of ammonium nitrogen. The increase in the UQ content was recognized at higher concentrations of 2, 4-d, especially with lower concentrations of sucrose. Physical factors had no marked effect on UQ formation except temperature. The UQ content was considerably raised at higher temperatures.     

微生物发酵法生产辅酶Q10研究进展

微生物发酵法是生产辅酶Q10很有前景的方法.本文综述了辅酶Q10产生菌的种类、生物合成机制、辅酶Q10产生菌的改良以及发酵条件优化等方面的研究进展.